Integrated screening and cultivation optimization of food processing wastewater-derived microalgae for enhanced biomass and lipid production toward sustainable biodiesel

Integrated screening and cultivation optimization of food processing wastewater-derived microalgae for enhanced biomass and lipid production toward sustainable biodiesel | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Integrated screening and cultivation optimization of food processing wastewater-derived microalgae for enhanced biomass and lipid production toward sustainable biodiesel Samart Taikhao, Saranya Phunpruch This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9112207/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 11 You are reading this latest preprint version Abstract Cultivating microalgae in wastewater represents a sustainable strategy for generating lipid-rich biomass for biodiesel production while concurrently achieving wastewater remediation. In this study, green microalgae isolated from food industry wastewater were screened to identify strains with high lipid production potential. The most promising isolate was identified by morphological characteristics and 18S rRNA gene analysis as Coelastrella sp. RMUTSB 01. Comparative cultivation in three types of food industry wastewater showed that wastewater from a processed food production plant (PFPP) supported the highest biomass growth and lipid production. Optimization of wastewater concentration indicated that 60% (v/v) PFPP wastewater was optimal for biomass and lipid production. Further optimization demonstrated that pH 8, 25°C, and a light intensity of 100 µmol photons m⁻ 2 s⁻ 1 maximized growth, yielding biomass and lipid productivities of 70.66 ± 1.89 and 35.51 ± 1.16 mg L⁻ 1 day⁻ 1 , respectively. Scale-up cultivation in 2.0-L flasks further increased biomass productivity to 102.79 ± 3.60 mg L⁻ 1 day⁻ 1 , with a lipid productivity of 48.80 ± 3.00 mg L⁻ 1 day⁻ 1 , and lipid content of 47.47 ± 2.51%. Simultaneously, nutrient and organic pollutant removal efficiencies reached 81–96% for COD, TDS, NH 3 –N, NO 3 ⁻, and PO 4 3 ⁻. The resulting PUFA-rich fatty acid profile supports both biodiesel production and high-value PUFA applications, highlighting Coelastrella sp. RMUTSB 01 as a promising candidate for integrated wastewater-based lipid production within a sustainable biorefinery framework. Coelastrella Wastewater Biomass Lipid Biodiesel Figures Figure 1 Figure 2 Figure 3 Highlights • A lipid-rich microalga sp. RMUTSB 01 was isolated from food-industry wastewater. • PFPP wastewater enabled high biomass and lipid accumulation under optimized conditions. • Biomass productivity reached 102.79 mg L⁻ day⁻ with 47.47% lipid content. • Efficient wastewater remediation achieved 81–96% removal of nutrients and COD. • PUFA-rich lipids support both biodiesel production and high-value bioproducts. Introduction The growing demand for renewable energy and the global shift toward a circular bioeconomy have increased interest in sustainable biofuels as alternatives to fossil fuels (Santos et al. 2025 ). Among potential bioenergy sources, green microalgae are considered highly promising due to their rapid growth, high photosynthetic efficiency, and ability to accumulate lipids without competing for arable land or freshwater resources. Many green microalgae can also adapt to nutrient-limited and fluctuating environments, including industrial and municipal wastewaters. This adaptability makes them suitable for both biomass production and wastewater treatment (Xu et al. 2020 ; Suh et al. 2024 ). Various types of wastewater, such as municipal, agricultural, and agro-industrial effluents, have been explored as alternative culture media because they contain high levels of organic carbon, nitrogen, and phosphorus (Ummalyma et al. 2022 ). Food-processing wastewater is particularly attractive due to its continuous generation and enrichment with biodegradable nutrients derived from carbohydrate-, protein-, and lipid-based processing (Taikhao and Phunpruch 2022 ). However, microalgal growth and lipid production in wastewater depend on several interacting factors, including wastewater composition, concentration, pH, light intensity, and temperature (Arumugham et al. 2025 ; Politaeva et al. 2023 ). In addition, lipid productivity varies greatly among microalgal strains, as different species and strains possess distinct metabolic capacities for lipid accumulation. Therefore, identifying strains that are well adapted to wastewater environments is essential for efficient lipid production. In Thailand, only a limited number of studies have explored the isolation and application of indigenous green microalgae from industrial wastewater for lipid or biofuel production. For example, Yeesang ( 2018 ) isolated several oleaginous microalgae, including Botryococcus sp., Chlorella sp., Scenedesmus sp., Volvox sp., and Dunaliella sp., capable of simultaneous lipid production and wastewater remediation in food-processing effluents, while Whangchenchom et al. ( 2014 ) demonstrated the cultivation of Scenedesmus sp. using instant noodle factory wastewater. Food-industry effluents have also supported biomass and biohydrogen production by Chlorella vulgaris var. vulgaris TISTR 8261 (Taikhao and Phunpruch 2022 ). More recently, Reungsang and Plangklang ( 2023 ) reported lipid- or carbohydrate-rich biomass production from Coelastrella sp. KKU-P1 and Acutodesmus sp. KKU-P2 cultivated in cassava ethanol wastewater. Despite these studies, systematic strain screening and resource-recovery strategies remain limited relative to the rapid expansion of the Thailand’s food-processing sector. Large volumes of wastewater from dairy, frozen food, and processed food industries are typically treated without recovering valuable resources (Pan et al. 2022 ). As these wastewaters are rich in nutrients, they could serve as low-cost culture media, and microalgae isolated from such environments may be naturally adapted to high nutrient levels and variable water quality. Therefore, further screening and evaluation of wastewater-derived green microalgae in Thailand are needed to support sustainable biofuel production. In this study, a green microalgal strain with high biomass and lipid production potential, isolated from food-processing industrial wastewater, was selected and evaluated as a sustainable platform for lipid production. Cultivation conditions in food-processing plant wastewater were optimized to enhance growth and lipid accumulation, while pollutant removal efficiency was assessed under wastewater-based cultivation. In addition, the fatty acid composition and predicted biodiesel properties of the produced lipids were analyzed to evaluate their suitability for biofuel applications. This study aims to demonstrate the feasibility of integrating wastewater treatment with microalgal lipid production within a sustainable biorefinery framework. Materials and methods Collection of wastewater samples and isolation of green microalgae Wastewater samples were collected from food-processing plants located in Phra Nakhon Si Ayutthaya Province, Thailand: a dairy product production plant (DPPP) (14°21'30.5"N, 100°40'17.4"E), a frozen food production plant (FFPP) (14°15'24.4"N, 100°36'13.2"E), and a processed food production plant (PFPP) (14°19'32.7"N, 100°38'39.5"E). Samples were obtained from the equalization ponds of the biological wastewater treatment systems in April 2024 and transported to the laboratory for further analysis. Approximately 300 mL of wastewater was collected in sterile glass bottles and incubated at 25°C under a light intensity of 30 µmol photons m⁻ 2 s⁻ 1 for 7 days to promote algal enrichment. Following the enrichment period, wastewater samples were centrifuged at 5,000 × g for 5 min. The supernatant was discarded, and the resulting cell pellets were washed twice with sterile distilled water to remove residual impurities. The concentrated algal suspension was then spread onto Tris–acetate–phosphate (TAP) agar (Harris 1989 ) using the spread plate technique. The plates were incubated at 25°C under a light intensity of 30 µmol photons m⁻ 2 s⁻ 1 for 7 days. Individual green microalgal colonies were subsequently selected and purified through repeated streak plating to obtain axenic cultures. The pure isolates were maintained under the same cultivation conditions and used for subsequent experiments. Screening of green microalgal isolates for lipid production The purified green microalgal isolates were cultivated in 250 mL Erlenmeyer flasks containing 100 mL of TAP medium. Cultures were maintained at a constant temperature of 25°C under continuous illumination at a light intensity of 30 µmol photons m⁻ 2 s⁻ 1 , with orbital shaking at 120 rpm. The cultures were grown for 5 days to allow sufficient biomass accumulation. At the end of the cultivation period, algal cells were harvested by centrifugation at 7,000 × g for 10 min at 4°C. The collected biomass was subsequently used for lipid content determination, which served as the primary criterion for screening high-lipid-producing isolates. Morphological observation and molecular identification The morphological characteristics of the purified microalgal isolates were examined using a light microscope (BX51, Olympus, Tokyo, Japan). For molecular identification, genomic DNA was extracted from fresh algal biomass using the Wizard SV Genomic DNA Purification Kit (Promega, Madison, WI, USA). The 18S rRNA gene was amplified by PCR using green algal-specific primers, F-18S rRNA (5′-CTGCGAATGGCTCATTAAATC-3′) and R-18S rRNA (5′-AAGGCCAGGGACGTAATCAA-3′) (Taikhao 2020 ), with a miniPCR system (mini16, miniPCR bio™, Cambridge, MA, USA). PCR amplification was carried out in a 50-µL reaction mixture containing 2× Taq PCR MasterMix II (Tiangen, Beijing, China), 250 nmol of each primer, and 50 ng of template DNA. The thermal cycling program consisted of an initial denaturation at 94°C for 3 min, followed by 35 cycles of denaturation at 94°C for 30 s, annealing at 58°C for 30 s, and extension at 72°C for 60 s, with a final extension at 72°C for 5 min. The amplified PCR products were purified using the GenepHlow™ Gel/PCR Purification Kit (Geneaid, New Taipei City, Taiwan) and sequenced bidirectionally (Macrogen, Seoul, Korea). Sequence similarity analysis was performed using BLASTN against the NCBI nucleotide database. Multiple sequence alignment was conducted using ClustalW (Larkin et al. 2007 ), and phylogenetic analysis was performed using the Maximum Likelihood method implemented in MEGA version 12 (Kumar et al. 2024 ) to confirm the taxonomic position of the algal isolate. Cultivation of the selected green microalga in different wastewater sources The selected microalgal cells were harvested by centrifugation at 7,000 × g for 10 min at 20°C, washed twice, and resuspended in sterilized wastewater. The initial inoculum density was adjusted to an OD 750 of approximately 0.1 (≈ 7.2 × 10 4 cells mL⁻ 1 ). Cultures were incubated at 25°C under a light intensity of 30 µmol photons m⁻² s⁻¹ on an orbital shaker at 120 rpm for 20 days. Growth parameters, including specific growth rate and doubling time, were monitored throughout the cultivation period. At the end of the experiment, lipid content, biomass productivity, and lipid productivity were determined. The wastewater source that supported the highest growth and lipid production was selected for subsequent optimization experiments. Effects of wastewater concentration on growth and lipid production The selected wastewater was diluted with distilled water to final concentrations of 0, 20, 40, 60, 80, and 100% (v/v). All treatments were inoculated to an initial OD 750 of approximately 0.1 and cultivated for 20 days under identical culture conditions. Growth kinetics were monitored throughout the cultivation period, and biomass- and lipid-related parameters were analyzed at the end of the experiment. The concentration level that resulted in the highest biomass accumulation and lipid production was selected for subsequent studies. Effects of pH, temperature and light intensity on growth and lipid production To evaluate the effects of environmental factors on microalgal growth and lipid production, the initial pH of the culture medium was adjusted to 4, 5, 6, 7, 8, or 9 using 1 M HCl or 1 M NaOH prior to sterilization. Temperature effects were investigated at 20, 25, 30, and 35°C, while light intensity was varied at 0, 30, 50, 100, and 150 µmol photons m⁻ 2 s⁻ 1 . All experiments were conducted for 20 days with agitation at 120 rpm under otherwise identical culture conditions. Growth kinetics, biomass production, and lipid accumulation were evaluated at the end of the cultivation period. Scale-up experiment For scale-up cultivation, algal cells were transferred to 2.0-L Erlenmeyer flasks containing 1.8 L of wastewater, with the initial inoculum density adjusted to an OD 750 of approximately 0.1. Cultures were grown under the optimized pH, temperature, and light intensity conditions with continuous aeration. Filtered air was supplied at a flow rate of 0.5 L min⁻ 1 through a 0.2-µm inline air filter and a sandstone bubble diffuser to ensure uniform mixing and stable aeration. The optical cell density, biomass production, and lipid accumulation were monitored throughout the cultivation period. After cultivation, the treated wastewater was collected and analyzed for changes in physicochemical parameters to evaluate nutrient removal efficiency. In addition, the fatty acid composition and biodiesel properties of fatty acid methyl esters (FAMEs) were determined. Analytical methods Wastewater analysis The pH and dissolved oxygen (DO) were measured on-site using a portable multi-parameter meter (HQ40D, Hach, Loveland, CO, USA), while electrical conductivity (EC) was determined using a conductivity meter (F30 FiveEasy, Mettler-Toledo, Columbus, OH, USA). Prior to chemical analysis, the samples were filtered through Whatman No. 3 filter paper (Cytiva, Marlborough, MA, USA) to remove suspended solids. Chemical oxygen demand (COD) and total dissolved solids (TDS) were analyzed in accordance with APHA Standard Methods 5220 C and 2540 C, respectively (APHA 2017). Concentrations of ammonia nitrogen (NH 3 –N), nitrate (NO 3 ⁻), and orthophosphate (PO 4 3 ⁻) were determined spectrophotometrically following APHA Standard Methods 4500-NH 3 F, 4500-NO 3 ⁻ B, and 4500-P E (APHA 2017). Prior to use in cultivation experiments, the wastewater samples were adjusted to pH 7, sterilized by autoclaving at 121°C for 15 min, and stored under sterile conditions. Growth assessment and biomass dry weight analysis Microalgal growth was monitored by withdrawing 1 mL of culture at designated time intervals and measuring the optical density at 750 nm (OD 750 ) using a UV–visible spectrophotometer (P1 UV–Visible Spectrophotometer, MAPADA, Shanghai, China), following established procedures. Biomass dry weight was determined by filtering 10 mL of culture through a pre-weighed GF/C glass microfiber filter (47 mm diameter; Whatman, Maidstone, UK). The filters were rinsed twice with distilled water to remove residual salts, dried at 85°C for 16 h, cooled in a desiccator, and reweighed to determine the dry biomass. The specific growth rate, doubling time, and biomass productivity were calculated using standard equations described in previous studies (Taikhao and Phunpruch, 2025 ). Total lipid extraction Algal cells cultivated for 20 days were harvested by centrifugation at 7,000 × g for 10 min at 4°C and subsequently dried at 60°C for 24 h. Total lipids were extracted from the dried biomass using a modified single-step extraction method based on Axelsson and Gentili ( 2014 ). Briefly, 20–30 mg of dried biomass was mixed with 8 mL of chloroform–methanol (2:1, v/v), followed by the addition of 2 mL of 0.73% (w/v) NaCl solution to induce phase separation. After centrifugation at 7,000 × g for 5 min, the chloroform phase was collected. The extraction procedure was repeated five times, and the pooled organic phases were evaporated under vacuum. The remaining lipid residue was weighed. Lipid content was expressed as a percentage of dry cell weight, lipid yield was calculated as mg L⁻ 1 , and lipid productivity was determined as lipid yield per culture volume per day (mg L⁻ 1 day⁻ 1 ). Fatty acid profile analysis The fatty acid composition of the extracted lipids was determined at the Scientific Instrument Center, School of Science, King Mongkut’s Institute of Technology Ladkrabang (KMITL), Bangkok, Thailand. Lipid extracts were converted to fatty acid methyl esters (FAMEs) via acid-catalyzed transesterification using 5% (v/v) HCl in methanol at 85°C for 1 h, following the method of Thongtha et al. ( 2025a ). The resulting FAMEs were analyzed using a gas chromatography–mass spectrometry system (GC 6890N coupled with MS 5973, Agilent Technologies, Santa Clara, CA, USA). Injector and detector temperatures were set at 250°C and 270°C, respectively. A 1-µL sample was injected in split mode (50:1), using high-purity helium (99.999%) as the carrier gas at a constant flow rate of 1 mL min⁻ 1 . Fatty acids were identified by comparing their mass spectra with reference spectra in the WILEY 7n.1 mass spectral library. The relative abundance of each fatty acid was calculated based on its peak area as a percentage of the total chromatographic peak area. Biodiesel properties of fatty acid methyl esters Biodiesel-related properties of the produced fatty acid methyl esters were estimated based on their fatty acid composition. Parameters including the degree of unsaturation, saponification value, iodine value, cetane number, cold flow properties, oxidation stability, higher heating value, kinematic viscosity, and density were calculated using Biodiesel Analyzer software (version 1.1) following the method described by Talebi et al. ( 2014 ). Statistical analysis All experimental results are presented as the mean ± standard deviation (SD) of three independent replicates. Error bars in the figures represent the corresponding SD values. Statistical comparisons among treatments were performed using one-way analysis of variance (ANOVA), followed by Duncan’s multiple range test to determine significant differences among means. A significance level of p < 0.05 was applied for all analyses. Statistical analyses were conducted using IBM SPSS Statistics version 24.0 (IBM Corp., Armonk, NY, USA). Results Screening of green microalgal isolates for total lipid production A total of 28 green microalgal isolates were purified from wastewater samples and examined under a light microscope. Their morphological characteristics are shown in Supplementary Fig. 1. Isolates SB-01 to SB-08 were obtained from FFPP wastewater, whereas isolates SB-09 to SB-27 were isolated from PFPP wastewater. The remaining isolate, SB-28, originated from DPPP wastewater. The isolates were subsequently screened for total lipid production. The total lipid content ranged from less than 0.2% to 2.99% of dry weight (Fig. 1 ). Among the isolates, SB-01 from FFPP wastewater exhibited the highest lipid content with 2.99% of dry weight, followed by SB-03, SB-17, and SB-08 (Fig. 1 ). The lipid content of SB-01 was approximately 1.2–26 times higher than that of the other isolates, indicating considerable variability in lipid accumulation among the strains. Based on its superior lipid content, isolate SB-01 was selected for taxonomic identification and further optimization of lipid production under wastewater-based cultivation conditions. Microalgal identification The high-lipid-producing microalgal isolate SB-01 was identified using a combination of morphological characterization and molecular phylogenetic analysis. Light microscopy revealed that SB-01 consisted of unicellular, bright green cells with an elliptical to slightly ovoid shape, occurring either singly or in small, loosely associated aggregates (Fig. 2 A). The cells measured approximately 8–12 µm in diameter and contained a parietal chloroplast, consistent with the diagnostic morphological characteristics of the genus Coelastrella . For molecular identification, the 18S rRNA gene of SB-01 was successfully amplified and sequenced. The obtained sequence was deposited in the NCBI GenBank database under accession number PX631087.1. BLASTN analysis revealed high sequence similarity to previously reported Coelastrella species. Phylogenetic analysis based on the Maximum Likelihood method placed SB-01 within a well-supported clade comprising Coelastrella thermophila , Coelastrella aeroterrestrica , Coelastrella rubescens , and Coelastrella terrestris , clearly distinct from other chlorophyte genera (Fig. 2 B). Based on the combined morphological characteristics and molecular phylogenetic data, isolate SB-01 was identified as Coelastrella sp. RMUTSB 01. Chemical characteristics of wastewater sources The three wastewater sources exhibited distinct chemical profiles, reflecting differences in their respective industrial processing activities. The pH values varied among the sources: wastewater from FFPP was near neutral, whereas wastewater from DPPP and PFPP was more acidic (Table 1 ). Wastewater temperatures ranged from 33 to 36°C across all samples (Table 1 ). EC and TDS were relatively high in wastewater from DPPP and PFPP (Table 1 ), indicating high ionic strength and elevated concentrations of dissolved solids. DO levels were consistently low in all wastewater samples (Table 1 ), suggesting high organic loading. COD was highest in PFPP wastewater (Table 1 ), followed by DPPP and FFPP, indicating a greater abundance of biodegradable organic matter in processed food effluents. PFPP wastewater also exhibited the highest concentrations of NH 3 –N, NO 3 ⁻, and PO 4 3 ⁻, whereas DPPP and FFPP contained substantially lower nutrient levels (Table 1 ). From the results, PFPP wastewater demonstrated the highest organic and nutrient loads among the three sources and is therefore expected to be a suitable nutrient-rich medium for microalgal cultivation. Table 1 Chemical characteristics of wastewater sources collected from food processing industries in Phra Nakhon Si Ayutthaya Province, Thailand. All values are presented as mean ± standard deviation (SD) from three independent experimental replicates. Statistically significant differences among samples within the same row are indicated by different superscript letters at a 95% confidence level. Parameters Wastewater sources Dairy product production plant (DPPP) Frozen food production plant (FFPP) Processed food production plant (PFPP) pH 5.57 ± 0.02 b 6.48 ± 0.03 a 4.52 ± 0.02 c Temperature 36.00 ± 0.60 a 33.00 ± 0.60 b 36.00 ± 0.40 a EC (µs cm⁻ 1 ) 2,340.00 ± 5.60 a 257.00 ± 1.70 b 2,350.00 ± 7.50 a DO (mgO 2 L⁻ 1 ) 0.83 ± 0.03 a 0.96 ± 0.51 a 1.13 ± 0.51 a COD (mgO 2 L⁻ 1 ) 2,144.10 ± 20.30 b 1,184.38 ± 30.02 c 2,560.00 ± 40.61 a TDS (mg L⁻ 1 ) 1,600.00 ± 20.00 a 800.00 ± 10.00 c 1,400.00 ± 15.00 b NH 3 -N (mg L⁻ 1 ) 6.45 ± 0.12 c 11.20 ± 0.32 b 21.25 ± 0.56 a NO 3 − (mg L⁻ 1 ) 0.42 ± 0.22 b 0.68 ± 0.17 b 4.72 ± 0.32 a PO 4 3− (mg L⁻ 1 ) 2.02 ± 0.12 b 1.05 ± 0.02 c 4.29 ± 0.11 a Growth and lipid production of Coelastrella sp. RMUTSB 01 in different wastewater sources Growth of Coelastrella sp. RMUTSB 01 was determined by measuring OD₇₅₀ in all wastewater sources and compared with that in the control TAP medium, a standard medium for green algae. Among the wastewater samples tested, Coelastrella sp. RMUTSB 01 demonstrated the highest growth in PFPP wastewater, with a specific growth rate of 0.42 ± 0.03 day − 1 , biomass productivity of 44.58 ± 2.65 mg L − 1 day − 1 , and the shortest doubling time of 1.64 ± 0.09 days, followed by FFPP and DPPP wastewater (Table 2 ). However, growth in PFPP was still lower than that observed in TAP medium (Table 2 ). In addition, Coelastrella sp. RMUTSB 01 cells cultivated in PFPP wastewater exhibited the highest lipid content of 20.94 ± 0.32% of dry weight and lipid productivity of 9.34 ± 0.77 mg L⁻ 1 day⁻ 1 (Table 2 ). In contrast, the lowest lipid content was shown in TAP medium (Table 2 ). It seems that growth and lipid production appeared to show an inverse relationship in TAP medium whereas in PFPP both parameters increased in the same direction. Based on these findings, PFPP wastewater was selected for subsequent optimization of lipid production in Coelastrella sp. RMUTSB 01. Table 2 Specific growth rate, doubling time, lipid content, biomass productivity, and lipid productivity of Coelastrella sp. RMUTSB 01 cultivated in DPPP, FFPP, and PFPP wastewater compared with standard TAP medium over 20 days. All values are presented as mean ± standard deviation (SD) from three independent experimental replicates. Statistically significant differences among samples within the same column are indicated by different superscript letters at a 95% confidence level. Culture media Specific growth rate (day⁻ 1 ) Biomass productivity (mg L⁻ 1 day⁻ 1 ) Doubling time (day) Lipid content (%) Lipid productivity (mg L⁻ 1 day⁻ 1 ) DPPP 0.35 ± 0.01 d 22.62 ± 2.15 d 1.98 ± 0.03 d 7.85 ± 0.25 c 1.78 ± 0.47 d FFPP 0.38 ± 0.02 c 38.05 ± 2.60 c 1.80 ± 0.02 c 12.11 ± 0.13 b 4.61 ± 0.89 b PFPP 0.42 ± 0.03 b 44.58 ± 2.65 b 1.64 ± 0.09 b 20.94 ± 0.32 a 9.34 ± 0.77 a TAP 0.46 ± 0.01 a 50.63 ± 2.95 a 1.50 ± 0.09 a 6.59 ± 0.16 d 3.34 ± 0.28 c Effects of PFPP wastewater concentration on growth and lipid production Due to its high ionic strength and elevated concentrations of dissolved solids, PFPP wastewater was diluted with autoclaved distilled water to obtain concentrations ranging from 0–100% for the enhancement of growth and lipid production. The highest growth of Coelastrella sp. RMUTSB 01 with a specific growth rate of 0.46 ± 0.02 day − 1 and biomass productivity of 51.03 ± 1.95 mg L − 1 day − 1 , was observed at 60% PFPP (Table 3 ). Both lower and higher concentrations than 60% resulted in reduced biomass productivity. Similarly, the maximum lipid content and lipid productivity of 37.06 ± 1.18% and 18.91 ± 1.61 mg L − 1 day − 1 , respectively, was shown at 60% PFPP wastewater (Table 3 ). Lipid productivity decreased at concentrations below or above 60%. These results indicate that 60% PFPP wastewater provided the most favorable nutrient composition suitable for both growth and lipid accumulation and was therefore selected for subsequent optimization experiments. Table 3 Specific growth rate, doubling time, lipid content, biomass productivity, and lipid productivity of Coelastrella sp. RMUTSB 01 cultivated under different concentrations of PFPP wastewater for 20 days. All values are presented as mean ± standard deviation (SD) from three independent experimental replicates. Statistically significant differences among samples within the same column are indicated by different superscript letters at a 95% confidence level. PFPP wastewater concentration (%) Specific growth rate (day⁻ 1 ) Biomass productivity (mg L⁻ 1 day⁻ 1 ) Doubling time (day) Lipid content (%) Lipid productivity (mg L⁻ 1 day⁻ 1 ) 0 0.08 ± 0.01 d 1.23 ± 0.04 e 8.18 ± 0.02 d 2.45 ± 0.13 f 0.03 ± 0.00 e 20 0.34 ± 0.03 c 22.71 ± 2.21 d 2.02 ± 0.04 c 11.49 ± 0.56 e 2.61 ± 0.62 d 40 0.38 ± 0.03 bc 30.62 ± 1.61 c 1.81 ± 0.07 b 31.31 ± 0.98 b 9.59 ± 1.13 c 60 0.46 ± 0.02 a 51.03 ± 1.95 a 1.52 ± 0.09 a 37.06 ± 1.18 a 18.91 ± 1.61 a 80 0.44 ± 0.04 a 46.08 ± 1.39 b 1.58 ± 0.08 a 28.33 ± 0.98 c 13.06 ± 1.62 b 100 0.42 ± 0.02 ab 44.58 ± 1.61 b 1.64 ± 0.10 a 20.94 ± 0.32 d 9.34 ± 1.28 c Effects of pH, temperature, and light intensity on growth and lipid production At temperature of 25°C and a light intensity at 30 µmol photons m⁻ 2 s⁻ 1 , pH strongly influenced the growth of Coelastrella sp. RMUTSB 01 in 60% PFPP wastewater. Increasing the pH from 4 to 8 enhanced growth and lipid production (Table 4 ).The maximum biomass productivity of 60.66 ± 1.39 mg L⁻ 1 day⁻ 1 and lipid productivity of 28.06 ± 1.60 mg L⁻ 1 day⁻ 1 was found at pH 8. A further increase to pH 9 resulted in a significant decline in these parameters (Table 4 ). Therefore, pH 8 was considered optimal for growth and lipid production in Coelastrella sp. RMUTSB 01. Table 4 Specific growth rate, doubling time, lipid content, biomass productivity, and lipid productivity of Coelastrella sp. RMUTSB 01 cultivated in 60% (v/v) PFPP wastewater under different pH, temperature, and light intensities for 20 days. All values are presented as mean ± standard deviation (SD) from three independent experimental replicates. Statistically significant differences among samples within the same column for each condition are indicated by different superscript letters at a 95% confidence level. pH Temperature ( o C) Light intensity (µmol photon m⁻ 2 s⁻ 1 ) Specific growth rate (day⁻ 1 ) Biomass productivity (mg L⁻ 1 day⁻ 1 ) Lipid content (%) Lipid productivity (mg L⁻ 1 day 1 ) 4 25 30 0.21 ± 0.02 d 21.23 ± 1.19 e 11.92 ± 0.13 e 2.53 ± 1.06 e 5 0.34 ± 0.02 c 27.71 ± 2.21 d 14.83 ± 0.56 d 4.11 ± 1.62 de 6 0.40 ± 0.03 b 35.62 ± 1.61 c 16.94 ± 0.98 d 6.03 ± 1.13 d 7 0.46 ± 0.02 ab 51.03 ± 1.95 b 37.06 ± 1.18 b 18.91 ± 1.61 b 8 0.48 ± 0.06 a 60.66 ± 1.39 a 46.25 ± 2.23 a 28.06 ± 1.60 a 9 0.42 ± 0.01 ab 49.58 ± 1.61 b 27.79 ± 1.79 c 13.78 ± 1.63 c 8 20 30 0.31 ± 0.04 c 37.71 ± 2.21 c 26.81 ± 1.12 d 10.11 ± 1.62 c 25 0.48 ± 0.06 a 60.66 ± 1.39 a 46.25 ± 2.23 a 28.06 ± 1.60 a 30 0.43 ± 0.03 a 45.62 ± 1.61 b 35.15 ± 1.44 b 16.03 ± 1.13 b 35 0.37 ± 0.03 b 44.58 ± 1.61 b 32.03 ± 1.25 c 14.28 ± 1.63 b 8 25 0 0.05 ± 0.02 b 1.28 ± 0.04 d 1.95 ± 0.01 c 0.03 ± 0.00 d 30 0.48 ± 0.06 a 60.66 ± 1.39 c 46.25 ± 2.23 b 28.06 ± 1.60 c 50 0.49 ± 0.02 a 64.62 ± 1.61 b 47.18 ± 1.29 ab 30.48 ± 1.63 bc 100 0.52 ± 0.03 a 70.66 ± 1.89 a 50.25 ± 2.18 a 35.51 ± 1.16 a 150 0.50 ± 0.04 a 65.58 ± 1.11 b 48.76 ± 2.05 ab 31.98 ± 2.13 b To evaluate the effect of incubation temperature, Coelastrella sp. RMUTSB 01 was cultivated at pH 8 and a light intensity of 30 µmol photons m⁻ 2 s⁻ 1 at temperature of 20, 25, 30, and 35°C. Cultivation at 25°C yielded the highest growth, as well as the highest biomass and lipid productivities (Table 4 ). Both lower (20°C) and higher temperatures (30–35°C) led to reduced growth and lipid production. Therefore, 25°C was selected for further optimization of light intensity. In Coelastrella sp. RMUTSB 01, negligible growth and lipid production were observed under dark conditions (Table 4 ). The highest growth with a specific growth rate of 0.52 ± 0.03 day − 1 , biomass productivity at 70.66 ± 1.89 mg L⁻ 1 day⁻1, lipid content of 50.25 ± 2.18%, and lipid productivity at 35.51 ± 1.16 mg L⁻ 1 day⁻ 1 , was achieved at a light intensity of 100 µmol photons m⁻ 2 s⁻ 1 (Table 4 ). These values decreased at 150 µmol photons m⁻ 2 s⁻ 1 , likely due to the onset of light saturation. In summary, pH 8, a temperature of 25°C, and a light intensity of 100 µmol photons m⁻ 2 s⁻ 1 were identified as the optimal conditions for biomass and lipid production in Coelastrella sp. RMUTSB 01 cultivated in 60% PFPP wastewater. Growth and lipid production by scale-up cultivation in 2.0-L Erlenmeyer flasks Growth and lipid production was further evaluated in Coelastrella sp. RMUTSB 01 cultivated in 2.0-L Erlenmeyer flasks containing 1.8 L of PFPP wastewater under two conditions: a control conditions (100% PFPP, pH 7, 25°C, 30 µmol photons m⁻ 2 s⁻ 1 ) and an optimal conditions (60% PFPP, pH 8, 25°C, 100 µmol photons m⁻ 2 s⁻ 1 ). Growth by OD 750 measurement under the optimal conditions was approximately 1.5-fold higher than those under the control conditions (Fig. 3 A). During 20 days of cultivation, biomass increased rapidly and reached a maximum on day 15 (Fig. 3 B). Under the optimal conditions, Coelastrella sp. RMUTSB 01 achieved the maximum biomass productivity of 102.79 ± 3.60 mg L⁻ 1 day⁻ 1 , which was about 1.5 times higher than that under the control conditions (68.91 ± 5.93 mg L⁻ 1 day⁻ 1 ). Moreover, lipid production of cells cultivated under the optimal conditions was approximately 3 fold higher that under the control conditions (Fig. 3 C). The maximum lipid content of 47.47 ± 2.51% and lipid productivity of 48.80 ± 3.00 mg L⁻ 1 day⁻ 1 was found on day 15 (Fig. 3 C), indicating a shift from active biomass production to lipid accumulation during the mid-stationary phase. Removal efficiencies of physicochemical parameters in PFPP wastewater The removal of physicochemical parameters from PFPP wastewater by Coelastrella sp. RMUTSB 01 cultivation under control and optimal conditions was evaluated. Substantial removal of all measured parameters was achieved under both conditions, with consistently higher removal efficiencies under the optimal conditions (Table 5 ). EC and COD were reduced by 85–87% and 78–81%, respectively, with slightly greater removal under the optimal conditions. Removal of TDS increased markedly under the optimal conditions, reaching 86%. Nitrogenous compounds were efficiently eliminated, with NH 3 –N and NO 3 ⁻ removal exceeding 90% under the optimal conditions. Similarly, phosphate removal improved to over 96% in the optimal conditions. These results indicate that cultivation of Coelastrella sp. RMUTSB 01, particularly under the optimal conditions, is highly effective for simultaneous nutrient removal and biomass/lipid production in PFPP wastewater. Table 5 Removal efficiencies (%) of physicochemical parameters in PFPP wastewater after cultivation of Coelastrella sp. RMUTSB 01 under control (100% PFPP, pH 7, 25°C, 30 µmol photons m⁻² s⁻¹) and optimal (60% PFPP, pH 8, 25°C, 100 µmol photons m⁻² s⁻¹) conditions. Data are presented as mean ± standard deviation (SD) (n = 3). Removal efficiencies (%) Control condition Optimal condition EC 85.10 ± 0.89 86.55 ± 0.34 COD 78.12 ± 0.65 81.46 ± 0.69 TDS 75.81 ± 2.21 86.44 ± 2.12 NH 3 -N 87.95 ± 2.89 91.06 ± 1.15 NO 3 − 86.22 ± 4.72 95.53 ± 0.46 PO 4 3− 85.54 ± 2.80 96.45 ± 0.50 Fatty acid composition and properties of biodiesel derived from lipids obtained under control and optimal conditions Coelastrella sp. RMUTSB 01 produced lipids dominated by polyunsaturated fatty acids (PUFA) under both conditions, with a higher PUFA proportion (70.8%) under the optimal conditions compared with the control conditions (65.7%) (Table 6 ). Linoleic acid (C18:2) and α-linolenic acid (C18:3) were the major PUFA detected under both conditions (Table 6 ). The contents of C16:2, C16:3, and eicosapentaenoic acid (EPA, C20:5) increased under the optimal conditions. In contrast, saturated fatty acids decreased from 31.4% to 25.8%, whereas monounsaturated fatty acids showed a slight increase under the optimal conditions (Table 6 ). Table 6 Fatty acid composition (% of total fatty acids) of lipids obtained from Coelastrella sp. RMUTSB 01 cells cultivated under control and optimal conditions for 20 days. Fatty acid Fatty acid composition (%) Control condition Optimal condition Myristic acid (C14:0) 0.6 0.5 Palmitic acid (C16:0) 27.3 23.1 Palmitoleic acid (C16:1) 0.8 1.2 Hexadecadienoic acid (C16:2) 9.0 11.8 Hexadecatrienoic acid (C16:3) 7.6 9.5 Stearic acid (C18:0) 3.6 2.2 Oleic acid (C18:1) 2.1 2.3 Linoleic acid (C18:2) 26.0 26.3 α-Linolenic acid (C18:3) 21.6 20.0 Eicosapentaenoic acid (C20:5) 1.4 3.1 Saturated fatty acid (SFA) 31.5 25.8 Monounsaturated fatty acid (MUFA) 2.9 3.5 Polyunsaturated fatty acid (PUFA) 65.6 70.7 Total identified fatty acids 100 100 The biodiesel properties estimated from the fatty acid profiles are shown in Table 7 . Under the optimal conditions, several fuel-related parameters improved, including an increase in the cetane number from 54.04 to 57.79, exceeding the minimum requirements of international biodiesel standards. Cold flow properties were also enhanced, as indicated by lower cold filter plugging point and cloud point values. Although oxidation stability increased slightly, it remained below standard requirements. The saponification and iodine values decreased under the optimal conditions but remained within acceptable ranges. In contrast, the kinematic viscosity, density, and higher heating value under both conditions were lower than standard biodiesel specifications. Therefore, the shift in fatty acid composition under the optimal conditions resulted in improved biodiesel-related properties, particularly with respect to ignition quality and low-temperature performance. Table 7. Properties of biodiesel derived from lipids obtained from Coelastrella sp. RMUTSB 01 cells cultivated under control and optimal conditions for 20 days. Biodiesel properties were predicted using the Biodiesel Analyzer (version 1.1). DU: Degree of unsaturation (% wt.), SV: Saponification value (mg g −1 ) IV: Iodine value g I 2 (100 g) −1 , CN: Cetane number, LCSF: Long-chain saturated factor, CFPP: Cold filter plugging point (°C), CP: Cloud point (°C), PP: Pour point (°C), APE: Allylic position equivalent, BAPE: Bis-allylic position equivalent, OS: Oxidation stability (h), HHV: Higher heating value (MJ kg −1 ), υ: Kinematic viscosity (mm 2 s −1 ), ρ: Density (kg m −3 ) Discussion In this study, screening of green microalgal isolates for lipid production revealed clear differences in lipid accumulation among the tested strains. Isolate SB-01 demonstrated the highest lipid content, approximately 1.2–26-fold higher than that of the other isolates (Fig. 1 ), highlighting substantial physiological variability among green microalgae isolated from the same or different industrial wastewater sources. Such variation is common in green microalgae and is associated with differences in carbon allocation, lipid biosynthetic capacity, and stress tolerance (Nayana et al. 2022 ). The high lipid-accumulating isolate SB-01 may be well adapted to elevated nutrient levels and capable of tolerating fluctuating environmental conditions. This adaptability likely enables SB-01 to produce biomass and accumulate lipids while simultaneously utilizing organic and inorganic compounds in wastewater for bioremediation. Comparable findings have been reported for green microalgae cultivated in sewage and laundry wastewater systems, where efficient removal of nutrients and organic loads was coupled with enhanced lipid accumulation and biodiesel-oriented biomass production (El Sheekh et al. 2023; Viena et al. 2025). The combined morphological and molecular analyses confirmed that isolate SB-01 belongs to the genus Coelastrella . Morphological observation showed that SB-01 consisted of unicellular, bright green cells with an elliptical to slightly ovoid shape and a parietal chloroplast, which are typical characteristic of this genus. The cell size ranged from 8–12 µm (Fig. 2 A), and the cells often formed loose aggregates. These features are similar to those reported for Coelastrella sp. UKM4 and Coelastrella S16/8 isolated from terrestrial and wastewater environments, respectively (Ding et al. 2020 ; Stamenov et al. 2025 ). Molecular phylogenetic analysis based on the 18S rRNA gene further supported this identification. Isolate SB-01 was placed within a well-supported Coelastrella clade that includes C. thermophila , C. aeroterrestrica , C. terrestris , and C. rubescens (Fig. 2 B). The sequence showed 99.57% similarity to C. thermophila var. globulina FACHB-2308. Phylogenetic analysis further placed the isolate within lineages containing known lipid- and pigment-producing Coelastrella strains (Wang et al. 2019 ). Recent taxonomic revisions have also clarified species boundaries within the genus Coelastrella , including the description of C. affinis sp. nov. and updates within the C. thermophila complex (Krivina et al. 2024 ). These studies highlight the high diversity within the genus, which also includes closely related taxa such as C. multistriata var. multistriata , C. striolata var. striolata , C. corcontica , and members of the C. rubescens lineage, together with C. oocystiformis and C. terrestris . Despite this diversity, many Coelastrella species share similar physiological traits that allow them to adapt to environmental stress. Species of Coelastrella are widely distributed in soils, aquatic systems, swamps, hot springs, and subaerial habitats, indicating strong ecological adaptability (Krivina et al. 2024 ). This adaptability is often linked to the accumulation of storage lipids and valuable metabolites such as carotenoids, including β-carotene and astaxanthin. For example, C. striolata var. multistriata strain 047 showed a biomass productivity of 26.67 mg L⁻ 1 day⁻ 1 and a lipid productivity of 6.71 mg L⁻ 1 day⁻ 1 under high-light conditions in humic-acid-containing medium (Susanti et al. 2024 ). Its fatty acid profile was mainly composed of oleic, α-linolenic, and palmitic acids. These findings support the identification of SB-01 as a Coelastrella strain and highlight its potential as a lipid-producing microalga isolated from food-processing wastewater. The wastewater from three sources (DPPP, FFPP, and PFPP) exhibited distinct physicochemical characteristics, reflecting differences in food-processing activities (Table 1 ). Such variation is commonly observed in industrial effluents and can strongly influence microalgal cultivation (Ansari et al. 2019 ). Among the tested sources, PFPP wastewater contained the highest organic and nutrient loads, as indicated by elevated COD, TDS, NH 3 –N, NO 3 ⁻, and PO 4 3 ⁻ concentrations (Table 1 ). Wastewater from processed food industries is typically rich in carbohydrates, proteins, and lipids. Consequently, it often contains high levels of organic carbon and nitrogen, which can support microalgal growth without the need for additional nutrient supplementation (Taikhao and Phunpruch 2025 ; Ummalyma et al. 2022 ). PFPP contained NH 3 -N at 21.25 ± 0.56 mg L⁻ 1 and PO 4 3 ⁻ at 4.29 ± 0.11 mg L⁻ 1 (Table 1 ). These values were higher than those measured in the other wastewater sources but substantially lower than the nutrient concentrations in the standard TAP medium (NH 3 -N 104.7 mg L⁻¹ and PO 4 3 ⁻ 47.47 mg L⁻¹). This intermediate nutrient level likely supported active growth while imposing moderate nitrogen and phosphorus limitation. Nitrogen availability plays a key role in regulating lipid metabolism. Under nitrogen-replete conditions, microalgae typically prioritize protein synthesis and biomass accumulation. In contrast, partial nitrogen limitation redirects carbon flux toward the synthesis of neutral lipids, particularly triacylglycerols (Maltsev et al. 2023 ). Phosphorus limitation can further reduce phospholipid and nucleic acid synthesis, thereby promoting neutral lipid accumulation as an alternative carbon storage pathway (Maltsev et al. 2023 ). Therefore, the relatively lower nitrogen and phosphorus levels in PFPP compared with TAP medium may create conditions that support Coelastrella sp. RMUTSB 01 growth while simultaneously enhancing lipid accumulation. Although the nutrient-rich TAP medium produced the highest specific growth rate and biomass productivity, lipid productivity remained relatively low (Table 2 ), indicating that these conditions primarily promoted cell proliferation rather than lipid accumulation. Among wastewater sources, PFPP wastewater showed higher biomass, lipid content and lipid productivity than other sources (Table 2 ). This can be attributed that PFPP wastewater contains higher NH 3 -N and PO 4 3 ⁻ concentrations compared with the other wastewaters, providing sufficient nutrients to support active microalgal growth. Biomass productivity in PFPP wastewater was higher than in FFPP and DPPP (Table 2 ) and exceeded values reported for Coelastrella sp. and Scenedesmus dimorphus cultivated in municipal wastewater (Lage and Gentili 2018 ). Similar trends have been reported for other microalgae grown in organic-rich food-industry wastewaters, including Chlorella sorokiniana , Coelastrella sp., and Chlorella pyrenoidosa cultivated in palm oil mill effluent (Ding et al. 2020 ). The lipid content obtained in PFPP wastewater (20.94%) was also substantially higher than values reported for Coelastrella sp. in municipal wastewater (< 10%) (Lage and Gentili 2018 ). In contrast, DPPP and FFPP wastewaters, which contain lower nutrient levels, resulted in lower growth and lipid production. This agrees with previous studies showing that dairy and low-strength food-processing wastewaters may limit nutrients or contain substances that reduce microalgal growth (Taikhao and Phunpruch 2025 ). Interestingly, Coelastrella sp. RMUTSB 01 was originally obtained from FFPP wastewater but exhibited significantly improved growth and lipid accumulation when cultivated in PFPP wastewater. One possible explanation is that FFPP wastewater originates from a frozen fried-food processing facility and may therefore contain relatively high levels of residual oils and lipids. Such conditions could favor the occurrence or selection of microalgae with strong lipid-accumulating capability. However, the nutrient composition of FFPP wastewater may not be optimal for sustained growth. In contrast, PFPP wastewater likely provides a more balanced nutrient environment that supports both active biomass production and lipid synthesis. These results therefore suggest that PFPP wastewater represents a suitable and low-cost cultivation medium for lipid-oriented microalgal production. By studying the responses of Coelastrella sp. RMUTSB 01 in terms of biomass and lipid production, it was found that nutrient levels and organic load strongly influence algal growth and lipid accumulation in PFPP wastewater. The lowest growth at 0% PFPP wastewater indicates that Coelastrella sp. RMUTSB 01 requires nitrogen and phosphorus for growth and metabolism, including photosynthesis and cell division (Table 3 ). Moderate concentrations at 20–40% PFPP wastewater sufficiently supported algal growth. At 60% PFPP wastewater, the highest growth rate, biomass productivity, lipid content, and lipid productivity were observed (Table 3 ). This suggests that 60% PFPP wastewater provides a suitable balance between sufficient nutrients and moderate metabolic stress, which can stimulate lipid accumulation. A similar dilution-optimization concept was reported by Suh et al. ( 2024 ), where Coelastrella sp. KNUA068 showed improved biomass in highly diluted cattle wastewater (1/50 diluted cattle wastewater, DCW). Likewise, Chlorella vulgaris and Scenedesmus sp. typically exhibited the highest biomass productivity at 25–50% wastewater strength in industrial sewage wastewater, while higher concentrations tend to reduce biomass productivity (Singh and Rathilal, 2024 ). At higher concentrations (80–100% PFPP wastewater), growth and lipid production decreased compared with the 60% PFPP wastewater, although they remained higher than in the low-strength cultures. This reduction is likely due to excessive organic matter, high ammonium levels, and increased TDS and EC, which can cause osmotic stress, oxidative stress, and pH imbalance within the cells. Similar inhibitory effects under highly concentrated wastewater conditions have been reported for C. striolata and Chlorella sorokiniana UKM3 cultivated in organic-rich effluents (Khalid et al. 2018 ; Susanti et al. 2024 ). Interestingly, the 60% PFPP concentration showed a growth rate similar to that of TAP medium but produced more than five times higher lipid productivity. This suggests that PFPP wastewater not only supplies essential nutrients but also provides additional organic substrates and micronutrients that are absent in defined media, while creating conditions that stimulate lipid biosynthesis in Coelastrella . The growth and lipid productivity of Coelastrella sp. RMUTSB 01 cultivated in PFPP wastewater were strongly influenced by pH, temperature, and light intensity. Biomass and lipid accumulation gradually increased from pH 4 to pH 8, with the maximum values observed at pH 8 (Table 4 ). This indicates that the strain prefers mildly alkaline conditions. This result agrees with previous studies on many green microalgae, such as Auxenochlorella protothecoides KP7, Coelastrella sp., Nannochloropsis salina , and Tetraselmis suecica , where neutral to slightly alkaline pH improves inorganic carbon availability and photosynthetic efficiency (Andeden et al. 2021 ; Singh et al. 2023 ; Arumugham et al. 2025 ). The superior biomass and lipid production at pH 8 in Coelastrella sp. RMUTSB 01 is also consistent with observations in C. rubescens , in which slightly alkaline pH enhanced both biomass production and lipid synthesis (Minyuk et al. 2016 ). Low growth at acidic pH values (4–5) is likely due to proton stress and reduced nutrient uptake. In contrast, the decrease in growth at pH 9 suggests alkaline stress and lower CO 2 availability, which has also been reported for Coelastrella cultivated in unbuffered wastewater systems (Zhou et al. 2017 ; Zeng et al. 2011 ). Temperature clearly affected growth and lipid production, with the highest biomass and lipid productivity observed at 25°C. This temperature falls within the typical mesophilic range for most green microalgae, where enzyme activity and carbon fixation operate most efficiently (Farrelly et al. 2013 ; Politaeva et al. 2023 ). Lower productivity at 20°C can be explained by reduced enzyme activity at low temperatures. In contrast, the decrease at high temperatures is likely due to heat stress, reduced CO₂ solubility, and damage to photosystem II (Zeng et al. 2011 ). Light intensity also strongly influenced growth and lipid accumulation. Biomass and lipid accumulation increased progressively from 30 to 100 µmol photons m⁻ 2 s⁻ 1 , with a clear optimum at 100 µmol photons m⁻ 2 s⁻ 1 . This reflects the classical saturation kinetics of photosynthesis, in which increasing irradiance enhances electron transport, ATP/NADPH generation, and carbon assimilation (Souliès et al. 2016 ). The strong enhancement of lipid accumulation at moderate irradiance (80–100 µmol photons m⁻ 2 s⁻ 1 ) is consistent with observations in Isochrysis galbana LB987 (Gim et al. 2016 ). However, increasing the light intensity to 150 µmol photons m⁻² s⁻¹ reduced productivity in Coelastrella sp. RMUTSB 01, likely due to photoinhibition and decreased of CO 2 fixation (Ashour et al. 2024 ). Similar effects have also been reported in Dunaliella salina and Nannochloropsis oculata CCAP849/1, where exposure to excessive light (200 µmol photons m⁻ 2 s⁻ 1 ) reduced lipid content (Gim et al. 2016 ). Scale-up cultivation in 2.0-L flasks confirmed that Coelastrella sp. RMUTSB 01 can maintain strong growth and lipid production under the optimal conditions (60% PFPP wastewater, pH 8, 25°C, 100 µmol photons m⁻ 2 s⁻ 1 ). Biomass reached a maximum of 1,542 mg L⁻ 1 on day 15, while lipid accumulation increased later, reaching 732 mg L⁻ 1 on day 15 (Fig. 3 ). This pattern indicates a typical shift from active growth to lipid accumulation during the stationary phase. Such delayed lipid production is commonly observed in Coelastrella sp. QY01 and Coelastrella sp. KNUA068, where cells initially prioritize biomass formation and later redirect carbon toward lipid storage under nutrient limitation or metabolic stress (Luo et al. 2016 ; Suh et al. 2024 ). Comparative analysis demonstrates that Coelastrella sp. RMUTSB 01 performs well compared with other green microalgae grown in wastewater (Table 8 ). The biomass yield of 1,542 mg L⁻ 1 and productivity of 102.79 mg L⁻ 1 day⁻ 1 after 15 days of cultivation were comparable to or higher than those reported for Coelastrella cultivated in cattle and municipal wastewaters (Ferro et al. 2018 ; Suh et al. 2024 ). These values were clearly higher than those obtained in humic-rich systems and palm oil mill effluent (POME) (Ding et al. 2020 ; Susanti et al. 2024 ; Lee et al. 2021 ). Table 8 Comparison of biomass yield, biomass productivity, lipid yield, and lipid productivity of Coelastrella sp. RMUTSB 01 compared with other Coelastrella strains cultivated in wastewater under different conditions. Green microalgal species Wastewater source Culture condition Biomass yield (mg L⁻ 1 ) Biomass productivity (mg L⁻ 1 day⁻ 1 ) Lipid yield (mg L⁻ 1 ) Lipid productivity (mg L⁻ 1 day⁻ 1 ) Lipid content (%) References Coelastrella sp. RMUTSB 01 Processed food production plant wastewater 2.0-L Erlenmeyer flask, sterile air, 60% wastewater, pH 8, temperature at 25°C and light intensity of 100 µmol photon m⁻ 2 s⁻ 1 1,542 102.79 732.00 48.80 47.47 This study Coelastrella sp. KNUA068 Cattle wastewater 250 mL Erlenmeyer flask, shaking at 160 rpm, 100% wastewater, temperature at 25°C and light intensity of 7 photon m⁻ 2 s⁻ 1 1,470 147.00 333.10 33.31 22.66 Suh et al ( 2024 ) Coelastrella striolata var. multistriata strain 047 Water-soluble humic acid (WSHA) 75% (v/v) WASHA at room temperature under continuous aeration and illumination at 20,000–25,000 lux 170 26.67 46.97 6.71 25.67 Susanti et al ( 2024 ) Coelastrella sp. KKU-P1 Unhydrolyzed molasses 1.0-L PBR, ambient concentration of CO 2 , BBM medium supplemented with molasses (10 g L⁻ 1 ) and a sodium nitrate (1.5 g L⁻ 1 ), pH 5, temperature at 25°C, continuous light illumination at 23.7 W m⁻ 2 4,280 305.71 - - 15.10 Thepsuthammarat et al ( 2023 ) Coelastrella sp. KJ-04 Distillery wastewater Distillery spent wash, shaking at 100 rpm, temperature at 25°C and light intensity of 70 µmol photon m⁻ 2 s⁻ 1 4,610 - 3,600 - - Vasistha et al (2023) Coelastrella sp. Piggery wastewater 0.5-L Cylinder-type PBR, 5% CO 2 , raw piggery wastewater, temperature at 25°C and light intensity of 1,250 µmol m⁻ 2 s⁻ 1 7,000–7,900 1,975 - 450 40 Lee et al ( 2021 ) Coelastrella sp. UKM4 Palm oil mill effluent (POME) Flask containing 1.8 L of pre-treated POME with 1% CO 2 mixed with air, temperature at 25°C and continuous light at 20,000 lux 930 187.20 - - - Ding et al ( 2020 ) Coelastrella sp. GN12 Energy grass digestates Bubbling column photobioreactors, 25% energy grass digestate: BG-11 (v: v) > 6,000 - 4,940 - > 50.00 Xu et al ( 2020 ) Coelastrella sp. (3–4) Municipal wastewater Multi-Cultivator MC 1000-OD photobioreactor, municipal wastewater at 25°C and continuous light intensity of 100 µmol photon m⁻ 2 s⁻ 1 1,460 0.11 450.00 34.61 30.80 Ferro et al ( 2018 ) Coelastrella sp. QY01 Aerobically treated swine wastewater (AnATSW) 0.5-L Erlenmeyer flask, 40% AnATSW, 25°C and light intensity of 100 µmol photon m⁻ 2 s⁻ 1 625 57.46 237.50 13.42 24.8 Luo et al ( 2016 ) Importantly, the lipid content of 47.47%, lipid yield of 732 mg L⁻ 1 , and lipid productivity of 48.80 mg L⁻ 1 day⁻ 1 after 15 days of cultivation were higher than most values reported for Coelastrella grown in wastewater, including cattle, swine, municipal, and molasses-based effluents (Luo et al. 2016 ; Ferro et al. 2018 ; Thepsuthammarat et al. 2023 ; Suh et al. 2024 ). Although some studies have reported higher lipid yields in high-strength wastewaters such as distillery effluent and digestate (Xu et al. 2020 ; Vasistha et al. 2023 ), these systems usually contain very high organic and nitrogen loads, which can create operational challenges. In comparison, PFPP wastewater enabled Coelastrella sp. RMUTSB 01 to achieve both high biomass productivity and high lipid accumulation under moderate-strength conditions. This balanced performance suggests that PFPP wastewater can serve as a practical and low-cost culture medium, highlighting the strong potential of Coelastrella sp. RMUTSB 01 for large-scale, lipid-focused microalgal cultivation within circular bioeconomy systems. The high removal efficiencies of EC, COD, TDS, NH 3 –N, NO 3 ⁻, and PO 4 3 ⁻ under optimal conditions demonstrate that Coelastrella sp. RMUTSB 01 is highly effective in treating nutrient-rich food-processing wastewater. The removal of nitrogen (91–96%) and phosphate (about 96%) indicates rapid uptake and incorporation into algal biomass. This confirms the dual function of the strain as wastewater treatment and biomass production. Similarly, strains such as Coelastrella sp. UKM4 and C. terrestris have shown the ability to survive and actively remove nutrients from challenging waste streams such as palm oil mill effluent (Badar et al. 2017 ; Ding et al. 2020 ; Udaiyappan et al. 2020 ) and municipal or industrial effluents (Al Raie et al., 2020 ). Cultivation of Coelastrella sp. GN12 in energy-grass digestate also demonstrated removal efficiencies exceeding 70% for nitrogen and 90% for phosphorus, further reinforcing the robustness of this genus in nutrient-rich environments (Xu et al. 2020 ). The strong remediation ability observed in this study is consistent with the known characteristics of the genus, such as efficient nitrogen assimilation, rapid phosphate uptake, and tolerance to high organic and salt concentrations. Improved nutrient removal under optimal pH, temperature, and light conditions also shows the close relationship between photosynthesis, biomass growth, and nutrient uptake, as previously described in carbon-rich wastewater systems (Vignesh et al. 2020 ). These results demonstrate that Coelastrella sp. RMUTSB 01 can achieve both high lipid production and efficient nutrient removal, making it a strong candidate for integrated wastewater treatment and value-added biomass production. The optimized cultivation conditions induced a significant shift in the fatty acid composition of Coelastrella sp. RMUTSB 01. The proportion of saturated fatty acids (SFAs) decreased from 31.5% to 25.8%, primarily due to reductions in palmitic acid (C16:0) and stearic acid (C18:0). In contrast, polyunsaturated fatty acids (PUFAs) increased from 65.6% to 70.7%, mainly driven by higher levels of C16:2, C16:3, C18:2, and C18:3, along with a more than twofold increase in eicosapentaenoic acid (EPA, C20:5). Comparable fatty acid remodeling has been reported in Coelastrella sp. V3 under a two-stage cultivation strategy, where an initial growth phase was followed by a modified second stage that stimulated lipid restructuring. This strategy led to pronounced changes in the proportions of C16:0, C18:0, C18:2, and C18:3 (Minhas et al., 2020 ). During the second stage, linoleic acid (C18:2) and α-linolenic acid (C18:3) became dominant components, significantly influencing the predicted biodiesel properties. The SFA-to-PUFA shift observed in the present study highlights the metabolic flexibility of Coelastrella , likely driven by enhanced desaturase activity and membrane lipid remodeling in response to the optimized cultivation conditions (Xu et al., 2020 ). Fatty acid composition is a key determinant of biodiesel quality, particularly the balance between saturated and unsaturated fatty acids (Minhas et al., 2016 ). Increased unsaturation generally improves cold-flow properties, whereas saturated fatty acids contribute to greater oxidative stability. In the present study, monounsaturated fatty acids (MUFAs) increased slightly from 2.92% to 3.43%. Despite this modest change, the presence of oleic acid (C18:1) remains beneficial, as this fatty acid is widely considered optimal for balancing ignition quality and oxidation resistance (Narayanan et al., 2018 ). These compositional changes translated into improved predicted fuel properties. The cetane number increased from 54.0 to 57.8, exceeding the minimum requirements specified by the EN 14214 standard and the Thailand DOEB guidelines. In addition, the cold filter plugging point decreased from − 2.28°C to − 5.83°C, indicating improved cold-flow performance. Similar enhancements in cold-flow behavior have been reported for PUFA-rich Coelastrella lipids, where increased unsaturation lowers crystallization temperature and improves low-temperature operability (Minhas et al., 2016 ). Therefore, while the fatty acid profile of Coelastrella sp. RMUTSB 01 supports acceptable ignition quality and favorable low-temperature performance, its high PUFA content may limit its direct biodiesel application as biodiesel without stabilization strategies, such as antioxidant supplementation or blending with more saturated feedstocks. At the same time, the elevated PUFA fraction particularly EPA (C20:5) and C18 polyunsaturated fatty acids enhances its potential for high-value applications, including nutraceutical and functional lipid production (Thongtha et al. 2025a and b ). Collectively, these findings highlight the dual potential of Coelastrella sp. RMUTSB 01 for integrated wastewater-based biomass production, offering flexibility toward either biodiesel generation or PUFA-rich lipid valorization depending on downstream processing priorities and market demand. Conclusion This study demonstrates that Coelastrella sp. RMUTSB 01 is a robust microalgal strain capable of simultaneously achieving efficient wastewater treatment and high lipid production. Among the tested substrates, PFPP wastewater significantly enhanced biomass and lipid accumulation compared with DPPP and FFPP. Under optimized conditions (60% PFPP, pH 8, 25°C, and 100 µmol photons m⁻² s⁻¹), the strain achieved a biomass productivity of 102.79 mg L⁻¹ day⁻¹, lipid content of 47.47%, and lipid productivity of 48.80 mg L⁻¹ day⁻¹, while removing 81–96% of nutrients and organic pollutants. The PUFA-rich fatty acid profile further indicates potential for both biodiesel production and high-value lipid applications. Therefore, Coelastrella sp. RMUTSB 01 shows strong promise for integrated biorefinery systems that convert agro-industrial wastewater into value-added biofuels and functional lipids, supporting circular bioeconomy strategies. Declarations Acknowledgements The authors appreciation to the three food industry factories in Phra Nakhon Si Ayutthaya Province for their kind cooperation in providing wastewater samples for this study. Authors’ contributions Conceptualization, S.T. and S.P.; methodology, S.T.; software, S.T.; validation, S.T. and S.P.; formal analysis, S.T. and S.P.; investigation, S.T.; resources, S.P.; data curation, S.T.; writing—original draft preparation, S.T. and S.P.; writing—review and editing, S.T. and S.P.; visualization, S.T.; supervision, S.P.; project administration, S.T., and S.P.; funding acquisition, S.T. and S.P. All authors have read and agreed to the published version of the manuscript. Funding This research did not receive funding. 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Udaiyappan AFM, Hasan HA, Takriff MS et al (2020) Microalgae–bacteria interaction in palm oil mill effluent treatment. J Water Process Eng 35:101203. Ummalyma SB, Sirohi R, Udayan A, Yadav P, Raj A, Sim SJ, Pandey A (2022) Sustainable microalgal biomass production in food industry wastewater for low-cost biorefinery products: a review. Phytochem Rev 22:969–991. Vasistha S, Balakrishnan D, Manivannan A, Rai MP (2023) Microalgae on distillery wastewater treatment. Chemosphere 315:137666. Viena V, Elvitriana, Juwandi, Suhendrayatna (2025) Effect of lighting cycles and wastewater concentrations on biomass growth and lipid content of green microalgae in laundry wastewater treatment. Results Eng 25:104316. Vignesh NS, Vimali E, Sangeetha R et al (2020) Sustainable biofuel from microalgae using lignocellulosic wastes. Fuel 278:118326. 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Supplementary Files SupplementaryFigure.docx Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Revision requested 27 Apr, 2026 Reviews received at journal 20 Apr, 2026 Reviews received at journal 18 Apr, 2026 Reviewers agreed at journal 01 Apr, 2026 Reviewers agreed at journal 30 Mar, 2026 Reviewers agreed at journal 28 Mar, 2026 Reviewers agreed at journal 27 Mar, 2026 Reviewers invited by journal 27 Mar, 2026 Editor assigned by journal 27 Mar, 2026 Submission checks completed at journal 24 Mar, 2026 First submitted to journal 13 Mar, 2026 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-9112207","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":614272862,"identity":"fe8d6a06-3728-4779-8cea-53cafc062f63","order_by":0,"name":"Samart Taikhao","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA6UlEQVRIiWNgGAWjYFAD9gYwlQAmGRsIKU9gkGDgOQBmQLQcJEqLRAKRWuTbTyd+rvzBUMc/8+3Tzbw/6vIY2A8/YP64A7cWgzO5myXPAG2RuJ1udpsn4XAxA0+aAcPBM3i0MORukGwAOex2GhtQy4HEBoYcoMPa8Dis/+3mnyAt8jePgbTUJTbwv8GvheFG7jawLQY32EBamBMbJAjYYnDj7TbLhjQJyY1n0thuzkk7XMwm8czgwFm8DsvdfLPBxoZf7vgxthtvbOry+PmTHz6oxOcwCJBAMNmA+ABBDaNgFIyCUTAK8AIA14VSY9GPO+oAAAAASUVORK5CYII=","orcid":"","institution":"Rajamangala University of Technology Suvarnabhumi","correspondingAuthor":true,"prefix":"","firstName":"Samart","middleName":"","lastName":"Taikhao","suffix":""},{"id":614272863,"identity":"e67ce5ee-7c26-476b-9798-1d970902db89","order_by":1,"name":"Saranya Phunpruch","email":"","orcid":"","institution":"King Mongkut's Institute of Technology Ladkrabang","correspondingAuthor":false,"prefix":"","firstName":"Saranya","middleName":"","lastName":"Phunpruch","suffix":""}],"badges":[],"createdAt":"2026-03-13 08:24:40","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-9112207/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-9112207/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":105815392,"identity":"df183380-b158-4455-a57f-8b281833d059","added_by":"auto","created_at":"2026-03-31 12:13:14","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":33940,"visible":true,"origin":"","legend":"\u003cp\u003eLipid content (% dry weight) of twenty-eight green microalgal strains isolated from wastewater of food processing plants in Phra Nakhon Si Ayutthaya Province, Thailand. The microalgal isolates were cultivated in TAP medium for 5 days at a constant temperature of 25 °C under continuous illumination at a light intensity of 30 µmol photons m⁻\u003csup\u003e2\u003c/sup\u003e s⁻\u003csup\u003e1\u003c/sup\u003e and shaking at 120 rpm. Bars represent mean values ± standard deviation (n = 3). Different letters above the bars indicate statistically significant differences among isolates (p \u0026lt; 0.05).\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-9112207/v1/8c452237cdf8fd69eecb2d67.png"},{"id":105815399,"identity":"eeb8ddcd-d54b-4f23-a519-abd557f754aa","added_by":"auto","created_at":"2026-03-31 12:13:20","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":598458,"visible":true,"origin":"","legend":"\u003cp\u003eMicroscopic morphology of \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 observed under light microscopy (scale bar = 10.0 µm) (A). Phylogenetic tree based on 18S rRNA gene sequences showing the relationship between \u003cem\u003eCoelastrella\u003c/em\u003esp. RMUTSB 01 and related species using the Maximum Likelihood method \u003cem\u003e(B)\u003c/em\u003e.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-9112207/v1/b8bb4d60a34839fdbefb0a65.png"},{"id":105815395,"identity":"7cafa357-ae2d-45b5-87de-5348b3ab8449","added_by":"auto","created_at":"2026-03-31 12:13:15","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":47366,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eComparison of growth by \u003c/strong\u003eoptical density at 750 nm (A)\u003cstrong\u003e, biomass yield (B), and lipid yield (C) of \u003c/strong\u003e\u003cem\u003eCoelastrella\u003c/em\u003e\u003cstrong\u003esp. RMUTSB 01 under control and optimal conditions \u003c/strong\u003eduring\u003cstrong\u003escaled-up cultivation for \u003c/strong\u003e20 days. Data are presented as mean ± standard deviation (SD) (n = 3). Different letters above the bars indicate statistically significant differences among isolates (p \u0026lt; 0.05).\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-9112207/v1/2ff7b8428f71e27f45846833.png"},{"id":105904623,"identity":"d2591b4e-2883-4d84-a30b-927d06fa4b1f","added_by":"auto","created_at":"2026-04-01 10:09:55","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2544930,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-9112207/v1/aa1bb9f9-6778-4729-b73a-43e02ca0d141.pdf"},{"id":105815394,"identity":"79873f7a-9299-4623-a66e-c75cff7106be","added_by":"auto","created_at":"2026-03-31 12:13:14","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":8408620,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryFigure.docx","url":"https://assets-eu.researchsquare.com/files/rs-9112207/v1/372b55a481769664abbc9455.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Integrated screening and cultivation optimization of food processing wastewater-derived microalgae for enhanced biomass and lipid production toward sustainable biodiesel","fulltext":[{"header":"Highlights","content":"\u003cp\u003e\u0026bull; A lipid-rich microalga sp. RMUTSB 01 was isolated from food-industry wastewater.\u003c/p\u003e\u003cp\u003e\u0026bull; PFPP wastewater enabled high biomass and lipid accumulation under optimized conditions.\u003c/p\u003e\u003cp\u003e\u0026bull; Biomass productivity reached 102.79 mg L⁻ day⁻ with 47.47% lipid content.\u003c/p\u003e\u003cp\u003e\u0026bull; Efficient wastewater remediation achieved 81\u0026ndash;96% removal of nutrients and COD.\u003c/p\u003e\u003cp\u003e\u0026bull; PUFA-rich lipids support both biodiesel production and high-value bioproducts.\u003c/p\u003e"},{"header":"Introduction","content":"\u003cp\u003eThe growing demand for renewable energy and the global shift toward a circular bioeconomy have increased interest in sustainable biofuels as alternatives to fossil fuels (Santos et al. \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). Among potential bioenergy sources, green microalgae are considered highly promising due to their rapid growth, high photosynthetic efficiency, and ability to accumulate lipids without competing for arable land or freshwater resources. Many green microalgae can also adapt to nutrient-limited and fluctuating environments, including industrial and municipal wastewaters. This adaptability makes them suitable for both biomass production and wastewater treatment (Xu et al. \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Suh et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2024\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eVarious types of wastewater, such as municipal, agricultural, and agro-industrial effluents, have been explored as alternative culture media because they contain high levels of organic carbon, nitrogen, and phosphorus (Ummalyma et al. \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Food-processing wastewater is particularly attractive due to its continuous generation and enrichment with biodegradable nutrients derived from carbohydrate-, protein-, and lipid-based processing (Taikhao and Phunpruch \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). However, microalgal growth and lipid production in wastewater depend on several interacting factors, including wastewater composition, concentration, pH, light intensity, and temperature (Arumugham et al. \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2025\u003c/span\u003e; Politaeva et al. \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). In addition, lipid productivity varies greatly among microalgal strains, as different species and strains possess distinct metabolic capacities for lipid accumulation. Therefore, identifying strains that are well adapted to wastewater environments is essential for efficient lipid production.\u003c/p\u003e \u003cp\u003eIn Thailand, only a limited number of studies have explored the isolation and application of indigenous green microalgae from industrial wastewater for lipid or biofuel production. For example, Yeesang (\u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e2018\u003c/span\u003e) isolated several oleaginous microalgae, including \u003cem\u003eBotryococcus\u003c/em\u003e sp., \u003cem\u003eChlorella\u003c/em\u003e sp., \u003cem\u003eScenedesmus\u003c/em\u003e sp., \u003cem\u003eVolvox\u003c/em\u003e sp., and \u003cem\u003eDunaliella\u003c/em\u003e sp., capable of simultaneous lipid production and wastewater remediation in food-processing effluents, while Whangchenchom et al. (\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e2014\u003c/span\u003e) demonstrated the cultivation of \u003cem\u003eScenedesmus\u003c/em\u003e sp. using instant noodle factory wastewater. Food-industry effluents have also supported biomass and biohydrogen production by \u003cem\u003eChlorella vulgaris\u003c/em\u003e var. \u003cem\u003evulgaris\u003c/em\u003e TISTR 8261 (Taikhao and Phunpruch \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). More recently, Reungsang and Plangklang (\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2023\u003c/span\u003e) reported lipid- or carbohydrate-rich biomass production from \u003cem\u003eCoelastrella\u003c/em\u003e sp. KKU-P1 and \u003cem\u003eAcutodesmus\u003c/em\u003e sp. KKU-P2 cultivated in cassava ethanol wastewater. Despite these studies, systematic strain screening and resource-recovery strategies remain limited relative to the rapid expansion of the Thailand\u0026rsquo;s food-processing sector. Large volumes of wastewater from dairy, frozen food, and processed food industries are typically treated without recovering valuable resources (Pan et al. \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). As these wastewaters are rich in nutrients, they could serve as low-cost culture media, and microalgae isolated from such environments may be naturally adapted to high nutrient levels and variable water quality. Therefore, further screening and evaluation of wastewater-derived green microalgae in Thailand are needed to support sustainable biofuel production.\u003c/p\u003e \u003cp\u003eIn this study, a green microalgal strain with high biomass and lipid production potential, isolated from food-processing industrial wastewater, was selected and evaluated as a sustainable platform for lipid production. Cultivation conditions in food-processing plant wastewater were optimized to enhance growth and lipid accumulation, while pollutant removal efficiency was assessed under wastewater-based cultivation. In addition, the fatty acid composition and predicted biodiesel properties of the produced lipids were analyzed to evaluate their suitability for biofuel applications. This study aims to demonstrate the feasibility of integrating wastewater treatment with microalgal lipid production within a sustainable biorefinery framework.\u003c/p\u003e"},{"header":"Materials and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eCollection of wastewater samples and isolation of green microalgae\u003c/h2\u003e \u003cp\u003eWastewater samples were collected from food-processing plants located in Phra Nakhon Si Ayutthaya Province, Thailand: a dairy product production plant (DPPP) (14\u0026deg;21'30.5\"N, 100\u0026deg;40'17.4\"E), a frozen food production plant (FFPP) (14\u0026deg;15'24.4\"N, 100\u0026deg;36'13.2\"E), and a processed food production plant (PFPP) (14\u0026deg;19'32.7\"N, 100\u0026deg;38'39.5\"E). Samples were obtained from the equalization ponds of the biological wastewater treatment systems in April 2024 and transported to the laboratory for further analysis. Approximately 300 mL of wastewater was collected in sterile glass bottles and incubated at 25\u0026deg;C under a light intensity of 30 \u0026micro;mol photons m⁻\u003csup\u003e2\u003c/sup\u003e s⁻\u003csup\u003e1\u003c/sup\u003e for 7 days to promote algal enrichment.\u003c/p\u003e \u003cp\u003eFollowing the enrichment period, wastewater samples were centrifuged at 5,000 \u0026times; g for 5 min. The supernatant was discarded, and the resulting cell pellets were washed twice with sterile distilled water to remove residual impurities. The concentrated algal suspension was then spread onto Tris\u0026ndash;acetate\u0026ndash;phosphate (TAP) agar (Harris \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e1989\u003c/span\u003e) using the spread plate technique. The plates were incubated at 25\u0026deg;C under a light intensity of 30 \u0026micro;mol photons m⁻\u003csup\u003e2\u003c/sup\u003e s⁻\u003csup\u003e1\u003c/sup\u003e for 7 days. Individual green microalgal colonies were subsequently selected and purified through repeated streak plating to obtain axenic cultures. The pure isolates were maintained under the same cultivation conditions and used for subsequent experiments.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eScreening of green microalgal isolates for lipid production\u003c/h3\u003e\n\u003cp\u003eThe purified green microalgal isolates were cultivated in 250 mL Erlenmeyer flasks containing 100 mL of TAP medium. Cultures were maintained at a constant temperature of 25\u0026deg;C under continuous illumination at a light intensity of 30 \u0026micro;mol photons m⁻\u003csup\u003e2\u003c/sup\u003e s⁻\u003csup\u003e1\u003c/sup\u003e, with orbital shaking at 120 rpm. The cultures were grown for 5 days to allow sufficient biomass accumulation. At the end of the cultivation period, algal cells were harvested by centrifugation at 7,000 \u0026times; g for 10 min at 4\u0026deg;C. The collected biomass was subsequently used for lipid content determination, which served as the primary criterion for screening high-lipid-producing isolates.\u003c/p\u003e\n\u003ch3\u003eMorphological observation and molecular identification\u003c/h3\u003e\n\u003cp\u003eThe morphological characteristics of the purified microalgal isolates were examined using a light microscope (BX51, Olympus, Tokyo, Japan). For molecular identification, genomic DNA was extracted from fresh algal biomass using the Wizard SV Genomic DNA Purification Kit (Promega, Madison, WI, USA). The 18S rRNA gene was amplified by PCR using green algal-specific primers, F-18S rRNA (5\u0026prime;-CTGCGAATGGCTCATTAAATC-3\u0026prime;) and R-18S rRNA (5\u0026prime;-AAGGCCAGGGACGTAATCAA-3\u0026prime;) (Taikhao \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), with a miniPCR system (mini16, miniPCR bio\u0026trade;, Cambridge, MA, USA). PCR amplification was carried out in a 50-\u0026micro;L reaction mixture containing 2\u0026times; \u003cem\u003eTaq\u003c/em\u003e PCR MasterMix II (Tiangen, Beijing, China), 250 nmol of each primer, and 50 ng of template DNA. The thermal cycling program consisted of an initial denaturation at 94\u0026deg;C for 3 min, followed by 35 cycles of denaturation at 94\u0026deg;C for 30 s, annealing at 58\u0026deg;C for 30 s, and extension at 72\u0026deg;C for 60 s, with a final extension at 72\u0026deg;C for 5 min. The amplified PCR products were purified using the GenepHlow\u0026trade; Gel/PCR Purification Kit (Geneaid, New Taipei City, Taiwan) and sequenced bidirectionally (Macrogen, Seoul, Korea). Sequence similarity analysis was performed using BLASTN against the NCBI nucleotide database. Multiple sequence alignment was conducted using ClustalW (Larkin et al. \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2007\u003c/span\u003e), and phylogenetic analysis was performed using the Maximum Likelihood method implemented in MEGA version 12 (Kumar et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2024\u003c/span\u003e) to confirm the taxonomic position of the algal isolate.\u003c/p\u003e\n\u003ch3\u003eCultivation of the selected green microalga in different wastewater sources\u003c/h3\u003e\n\u003cp\u003eThe selected microalgal cells were harvested by centrifugation at 7,000 \u0026times; g for 10 min at 20\u0026deg;C, washed twice, and resuspended in sterilized wastewater. The initial inoculum density was adjusted to an OD\u003csub\u003e750\u003c/sub\u003e of approximately 0.1 (\u0026asymp;\u0026thinsp;7.2 \u0026times; 10\u003csup\u003e4\u003c/sup\u003e cells mL⁻\u003csup\u003e1\u003c/sup\u003e). Cultures were incubated at 25\u0026deg;C under a light intensity of 30 \u0026micro;mol photons m⁻\u0026sup2; s⁻\u0026sup1; on an orbital shaker at 120 rpm for 20 days. Growth parameters, including specific growth rate and doubling time, were monitored throughout the cultivation period. At the end of the experiment, lipid content, biomass productivity, and lipid productivity were determined. The wastewater source that supported the highest growth and lipid production was selected for subsequent optimization experiments.\u003c/p\u003e\n\u003ch3\u003eEffects of wastewater concentration on growth and lipid production\u003c/h3\u003e\n\u003cp\u003eThe selected wastewater was diluted with distilled water to final concentrations of 0, 20, 40, 60, 80, and 100% (v/v). All treatments were inoculated to an initial OD\u003csub\u003e750\u003c/sub\u003e of approximately 0.1 and cultivated for 20 days under identical culture conditions. Growth kinetics were monitored throughout the cultivation period, and biomass- and lipid-related parameters were analyzed at the end of the experiment. The concentration level that resulted in the highest biomass accumulation and lipid production was selected for subsequent studies.\u003c/p\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eEffects of pH, temperature and light intensity on growth and lipid production\u003c/h2\u003e \u003cp\u003eTo evaluate the effects of environmental factors on microalgal growth and lipid production, the initial pH of the culture medium was adjusted to 4, 5, 6, 7, 8, or 9 using 1 M HCl or 1 M NaOH prior to sterilization. Temperature effects were investigated at 20, 25, 30, and 35\u0026deg;C, while light intensity was varied at 0, 30, 50, 100, and 150 \u0026micro;mol photons m⁻\u003csup\u003e2\u003c/sup\u003e s⁻\u003csup\u003e1\u003c/sup\u003e. All experiments were conducted for 20 days with agitation at 120 rpm under otherwise identical culture conditions. Growth kinetics, biomass production, and lipid accumulation were evaluated at the end of the cultivation period.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eScale-up experiment\u003c/h3\u003e\n\u003cp\u003eFor scale-up cultivation, algal cells were transferred to 2.0-L Erlenmeyer flasks containing 1.8 L of wastewater, with the initial inoculum density adjusted to an OD\u003csub\u003e750\u003c/sub\u003e of approximately 0.1. Cultures were grown under the optimized pH, temperature, and light intensity conditions with continuous aeration. Filtered air was supplied at a flow rate of 0.5 L min⁻\u003csup\u003e1\u003c/sup\u003e through a 0.2-\u0026micro;m inline air filter and a sandstone bubble diffuser to ensure uniform mixing and stable aeration. The optical cell density, biomass production, and lipid accumulation were monitored throughout the cultivation period. After cultivation, the treated wastewater was collected and analyzed for changes in physicochemical parameters to evaluate nutrient removal efficiency. In addition, the fatty acid composition and biodiesel properties of fatty acid methyl esters (FAMEs) were determined.\u003c/p\u003e\n\u003ch3\u003eAnalytical methods\u003c/h3\u003e\n\u003cp\u003eWastewater analysis\u003c/p\u003e \u003cp\u003eThe pH and dissolved oxygen (DO) were measured on-site using a portable multi-parameter meter (HQ40D, Hach, Loveland, CO, USA), while electrical conductivity (EC) was determined using a conductivity meter (F30 FiveEasy, Mettler-Toledo, Columbus, OH, USA). Prior to chemical analysis, the samples were filtered through Whatman No. 3 filter paper (Cytiva, Marlborough, MA, USA) to remove suspended solids. Chemical oxygen demand (COD) and total dissolved solids (TDS) were analyzed in accordance with APHA Standard Methods 5220 C and 2540 C, respectively (APHA 2017). Concentrations of ammonia nitrogen (NH\u003csub\u003e3\u003c/sub\u003e\u0026ndash;N), nitrate (NO\u003csub\u003e3\u003c/sub\u003e⁻), and orthophosphate (PO\u003csub\u003e4\u003c/sub\u003e\u003csup\u003e3\u003c/sup\u003e⁻) were determined spectrophotometrically following APHA Standard Methods 4500-NH\u003csub\u003e3\u003c/sub\u003e F, 4500-NO\u003csub\u003e3\u003c/sub\u003e⁻ B, and 4500-P E (APHA 2017). Prior to use in cultivation experiments, the wastewater samples were adjusted to pH 7, sterilized by autoclaving at 121\u0026deg;C for 15 min, and stored under sterile conditions.\u003c/p\u003e \u003cp\u003eGrowth assessment and biomass dry weight analysis\u003c/p\u003e \u003cp\u003eMicroalgal growth was monitored by withdrawing 1 mL of culture at designated time intervals and measuring the optical density at 750 nm (OD\u003csub\u003e750\u003c/sub\u003e) using a UV\u0026ndash;visible spectrophotometer (P1 UV\u0026ndash;Visible Spectrophotometer, MAPADA, Shanghai, China), following established procedures. Biomass dry weight was determined by filtering 10 mL of culture through a pre-weighed GF/C glass microfiber filter (47 mm diameter; Whatman, Maidstone, UK). The filters were rinsed twice with distilled water to remove residual salts, dried at 85\u0026deg;C for 16 h, cooled in a desiccator, and reweighed to determine the dry biomass. The specific growth rate, doubling time, and biomass productivity were calculated using standard equations described in previous studies (Taikhao and Phunpruch, \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2025\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eTotal lipid extraction\u003c/p\u003e \u003cp\u003eAlgal cells cultivated for 20 days were harvested by centrifugation at 7,000 \u0026times; g for 10 min at 4\u0026deg;C and subsequently dried at 60\u0026deg;C for 24 h. Total lipids were extracted from the dried biomass using a modified single-step extraction method based on Axelsson and Gentili (\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). Briefly, 20\u0026ndash;30 mg of dried biomass was mixed with 8 mL of chloroform\u0026ndash;methanol (2:1, v/v), followed by the addition of 2 mL of 0.73% (w/v) NaCl solution to induce phase separation. After centrifugation at 7,000 \u0026times; g for 5 min, the chloroform phase was collected. The extraction procedure was repeated five times, and the pooled organic phases were evaporated under vacuum. The remaining lipid residue was weighed. Lipid content was expressed as a percentage of dry cell weight, lipid yield was calculated as mg L⁻\u003csup\u003e1\u003c/sup\u003e, and lipid productivity was determined as lipid yield per culture volume per day (mg L⁻\u003csup\u003e1\u003c/sup\u003e day⁻\u003csup\u003e1\u003c/sup\u003e).\u003c/p\u003e \u003cp\u003eFatty acid profile analysis\u003c/p\u003e \u003cp\u003eThe fatty acid composition of the extracted lipids was determined at the Scientific Instrument Center, School of Science, King Mongkut\u0026rsquo;s Institute of Technology Ladkrabang (KMITL), Bangkok, Thailand. Lipid extracts were converted to fatty acid methyl esters (FAMEs) via acid-catalyzed transesterification using 5% (v/v) HCl in methanol at 85\u0026deg;C for 1 h, following the method of Thongtha et al. (\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2025a\u003c/span\u003e). The resulting FAMEs were analyzed using a gas chromatography\u0026ndash;mass spectrometry system (GC 6890N coupled with MS 5973, Agilent Technologies, Santa Clara, CA, USA). Injector and detector temperatures were set at 250\u0026deg;C and 270\u0026deg;C, respectively. A 1-\u0026micro;L sample was injected in split mode (50:1), using high-purity helium (99.999%) as the carrier gas at a constant flow rate of 1 mL min⁻\u003csup\u003e1\u003c/sup\u003e. Fatty acids were identified by comparing their mass spectra with reference spectra in the WILEY 7n.1 mass spectral library. The relative abundance of each fatty acid was calculated based on its peak area as a percentage of the total chromatographic peak area.\u003c/p\u003e \u003cp\u003eBiodiesel properties of fatty acid methyl esters\u003c/p\u003e \u003cp\u003eBiodiesel-related properties of the produced fatty acid methyl esters were estimated based on their fatty acid composition. Parameters including the degree of unsaturation, saponification value, iodine value, cetane number, cold flow properties, oxidation stability, higher heating value, kinematic viscosity, and density were calculated using Biodiesel Analyzer software (version 1.1) following the method described by Talebi et al. (\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2014\u003c/span\u003e).\u003c/p\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eStatistical analysis\u003c/h2\u003e \u003cp\u003eAll experimental results are presented as the mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation (SD) of three independent replicates. Error bars in the figures represent the corresponding SD values. Statistical comparisons among treatments were performed using one-way analysis of variance (ANOVA), followed by Duncan\u0026rsquo;s multiple range test to determine significant differences among means. A significance level of p\u0026thinsp;\u0026lt;\u0026thinsp;0.05 was applied for all analyses. Statistical analyses were conducted using IBM SPSS Statistics version 24.0 (IBM Corp., Armonk, NY, USA).\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec13\" class=\"Section2\"\u003e\n \u003ch2\u003eScreening of green microalgal isolates for total lipid production\u003c/h2\u003e\n \u003cp\u003eA total of 28 green microalgal isolates were purified from wastewater samples and examined under a light microscope. Their morphological characteristics are shown in Supplementary Fig. 1. Isolates SB-01 to SB-08 were obtained from FFPP wastewater, whereas isolates SB-09 to SB-27 were isolated from PFPP wastewater. The remaining isolate, SB-28, originated from DPPP wastewater. The isolates were subsequently screened for total lipid production. The total lipid content ranged from less than 0.2% to 2.99% of dry weight (Fig. \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Among the isolates, SB-01 from FFPP wastewater exhibited the highest lipid content with 2.99% of dry weight, followed by SB-03, SB-17, and SB-08 (Fig. \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The lipid content of SB-01 was approximately 1.2\u0026ndash;26 times higher than that of the other isolates, indicating considerable variability in lipid accumulation among the strains. Based on its superior lipid content, isolate SB-01 was selected for taxonomic identification and further optimization of lipid production under wastewater-based cultivation conditions.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e\n \u003ch2\u003eMicroalgal identification\u003c/h2\u003e\n \u003cp\u003eThe high-lipid-producing microalgal isolate SB-01 was identified using a combination of morphological characterization and molecular phylogenetic analysis. Light microscopy revealed that SB-01 consisted of unicellular, bright green cells with an elliptical to slightly ovoid shape, occurring either singly or in small, loosely associated aggregates (Fig. \u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eA). The cells measured approximately 8\u0026ndash;12 \u0026micro;m in diameter and contained a parietal chloroplast, consistent with the diagnostic morphological characteristics of the genus \u003cem\u003eCoelastrella\u003c/em\u003e. For molecular identification, the 18S rRNA gene of SB-01 was successfully amplified and sequenced. The obtained sequence was deposited in the NCBI GenBank database under accession number PX631087.1. BLASTN analysis revealed high sequence similarity to previously reported \u003cem\u003eCoelastrella\u003c/em\u003e species. Phylogenetic analysis based on the Maximum Likelihood method placed SB-01 within a well-supported clade comprising \u003cem\u003eCoelastrella thermophila\u003c/em\u003e, \u003cem\u003eCoelastrella aeroterrestrica\u003c/em\u003e, \u003cem\u003eCoelastrella rubescens\u003c/em\u003e, and \u003cem\u003eCoelastrella terrestris\u003c/em\u003e, clearly distinct from other chlorophyte genera (Fig. \u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eB). Based on the combined morphological characteristics and molecular phylogenetic data, isolate SB-01 was identified as \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec15\" class=\"Section2\"\u003e\n \u003ch2\u003eChemical characteristics of wastewater sources\u003c/h2\u003e\n \u003cp\u003eThe three wastewater sources exhibited distinct chemical profiles, reflecting differences in their respective industrial processing activities. The pH values varied among the sources: wastewater from FFPP was near neutral, whereas wastewater from DPPP and PFPP was more acidic (Table \u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Wastewater temperatures ranged from 33 to 36\u0026deg;C across all samples (Table \u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). EC and TDS were relatively high in wastewater from DPPP and PFPP (Table \u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e), indicating high ionic strength and elevated concentrations of dissolved solids. DO levels were consistently low in all wastewater samples (Table \u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e), suggesting high organic loading. COD was highest in PFPP wastewater (Table \u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e), followed by DPPP and FFPP, indicating a greater abundance of biodegradable organic matter in processed food effluents. PFPP wastewater also exhibited the highest concentrations of NH\u003csub\u003e3\u003c/sub\u003e\u0026ndash;N, NO\u003csub\u003e3\u003c/sub\u003e⁻, and PO\u003csub\u003e4\u003c/sub\u003e\u003csup\u003e3\u003c/sup\u003e⁻, whereas DPPP and FFPP contained substantially lower nutrient levels (Table \u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). From the results, PFPP wastewater demonstrated the highest organic and nutrient loads among the three sources and is therefore expected to be a suitable nutrient-rich medium for microalgal cultivation.\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\u0026nbsp;\u0026nbsp;\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eChemical characteristics of wastewater sources collected from food processing industries in Phra Nakhon Si Ayutthaya Province, Thailand. All values are presented as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation (SD) from three independent experimental replicates. Statistically significant differences among samples within the same row are indicated by different superscript letters at a 95% confidence level.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e\n \u003cp\u003eParameters\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"3\" nameend=\"c4\" namest=\"c2\"\u003e\n \u003cp\u003eWastewater sources\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003eDairy product production plant\u003c/p\u003e\n \u003cp\u003e(DPPP)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003eFrozen food production plant\u003c/p\u003e\n \u003cp\u003e(FFPP)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003eProcessed food production plant\u003c/p\u003e\n \u003cp\u003e(PFPP)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003epH\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e5.57\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e6.48\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e4.52\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eTemperature\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e36.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.60\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e33.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.60\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e36.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.40\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eEC (\u0026micro;s cm⁻\u003csup\u003e1\u003c/sup\u003e)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e2,340.00\u0026thinsp;\u0026plusmn;\u0026thinsp;5.60\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e257.00\u0026thinsp;\u0026plusmn;\u0026thinsp;1.70\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e2,350.00\u0026thinsp;\u0026plusmn;\u0026thinsp;7.50\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eDO (mgO\u003csub\u003e2\u003c/sub\u003e L⁻\u003csup\u003e1\u003c/sup\u003e)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e0.83\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e0.96\u0026thinsp;\u0026plusmn;\u0026thinsp;0.51\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e1.13\u0026thinsp;\u0026plusmn;\u0026thinsp;0.51\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eCOD (mgO\u003csub\u003e2\u003c/sub\u003e L⁻\u003csup\u003e1\u003c/sup\u003e)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e2,144.10\u0026thinsp;\u0026plusmn;\u0026thinsp;20.30\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e1,184.38\u0026thinsp;\u0026plusmn;\u0026thinsp;30.02\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e2,560.00\u0026thinsp;\u0026plusmn;\u0026thinsp;40.61\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eTDS (mg L⁻\u003csup\u003e1\u003c/sup\u003e)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e1,600.00\u0026thinsp;\u0026plusmn;\u0026thinsp;20.00\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e800.00\u0026thinsp;\u0026plusmn;\u0026thinsp;10.00\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e1,400.00\u0026thinsp;\u0026plusmn;\u0026thinsp;15.00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eNH\u003csub\u003e3\u003c/sub\u003e-N (mg L⁻\u003csup\u003e1\u003c/sup\u003e)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e6.45\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e11.20\u0026thinsp;\u0026plusmn;\u0026thinsp;0.32\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e21.25\u0026thinsp;\u0026plusmn;\u0026thinsp;0.56\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eNO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e (mg L⁻\u003csup\u003e1\u003c/sup\u003e)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e0.42\u0026thinsp;\u0026plusmn;\u0026thinsp;0.22\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e0.68\u0026thinsp;\u0026plusmn;\u0026thinsp;0.17\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e4.72\u0026thinsp;\u0026plusmn;\u0026thinsp;0.32\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003ePO\u003csub\u003e4\u003c/sub\u003e\u003csup\u003e3\u0026minus;\u003c/sup\u003e (mg L⁻\u003csup\u003e1\u003c/sup\u003e)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e2.02\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e1.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e4.29\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n \u003cp\u003e\u003cstrong\u003eGrowth and lipid production of\u003c/strong\u003e \u003cstrong\u003eCoelastrella\u003c/strong\u003e \u003cstrong\u003esp. RMUTSB 01 in different wastewater sources\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eGrowth of \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 was determined by measuring OD₇₅₀ in all wastewater sources and compared with that in the control TAP medium, a standard medium for green algae.\u003c/p\u003e\n \u003cp\u003eAmong the wastewater samples tested, \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 demonstrated the highest growth in PFPP wastewater, with a specific growth rate of 0.42\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03 day\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, biomass productivity of 44.58\u0026thinsp;\u0026plusmn;\u0026thinsp;2.65 mg L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e day\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, and the shortest doubling time of 1.64\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09 days, followed by FFPP and DPPP wastewater (Table \u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). However, growth in PFPP was still lower than that observed in TAP medium (Table \u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). In addition, \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 cells cultivated in PFPP wastewater exhibited the highest lipid content of 20.94\u0026thinsp;\u0026plusmn;\u0026thinsp;0.32% of dry weight and lipid productivity of 9.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.77 mg L⁻\u003csup\u003e1\u003c/sup\u003e day⁻\u003csup\u003e1\u003c/sup\u003e (Table \u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). In contrast, the lowest lipid content was shown in TAP medium (Table \u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). It seems that growth and lipid production appeared to show an inverse relationship in TAP medium whereas in PFPP both parameters increased in the same direction. Based on these findings, PFPP wastewater was selected for subsequent optimization of lipid production in \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01.\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\u0026nbsp;\u0026nbsp;\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eSpecific growth rate, doubling time, lipid content, biomass productivity, and lipid productivity of \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 cultivated in DPPP, FFPP, and PFPP wastewater compared with standard TAP medium over 20 days. All values are presented as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation (SD) from three independent experimental replicates. Statistically significant differences among samples within the same column are indicated by different superscript letters at a 95% confidence level.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eCulture media\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003eSpecific growth rate\u003c/p\u003e\n \u003cp\u003e(day⁻\u003csup\u003e1\u003c/sup\u003e)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003eBiomass productivity\u003c/p\u003e\n \u003cp\u003e(mg L⁻\u003csup\u003e1\u003c/sup\u003e day⁻\u003csup\u003e1\u003c/sup\u003e)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003eDoubling time\u003c/p\u003e\n \u003cp\u003e(day)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003eLipid content\u003c/p\u003e\n \u003cp\u003e(%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003eLipid productivity\u003c/p\u003e\n \u003cp\u003e(mg L⁻\u003csup\u003e1\u003c/sup\u003e day⁻\u003csup\u003e1\u003c/sup\u003e)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eDPPP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e0.35\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e22.62\u0026thinsp;\u0026plusmn;\u0026thinsp;2.15\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e1.98\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e7.85\u0026thinsp;\u0026plusmn;\u0026thinsp;0.25\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e1.78\u0026thinsp;\u0026plusmn;\u0026thinsp;0.47\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eFFPP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e0.38\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e38.05\u0026thinsp;\u0026plusmn;\u0026thinsp;2.60\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e1.80\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e12.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.13\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e4.61\u0026thinsp;\u0026plusmn;\u0026thinsp;0.89\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003ePFPP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e0.42\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e44.58\u0026thinsp;\u0026plusmn;\u0026thinsp;2.65\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e1.64\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e20.94\u0026thinsp;\u0026plusmn;\u0026thinsp;0.32\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e9.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.77\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eTAP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e0.46\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e50.63\u0026thinsp;\u0026plusmn;\u0026thinsp;2.95\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e1.50\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e6.59\u0026thinsp;\u0026plusmn;\u0026thinsp;0.16\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e3.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.28\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n \u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec16\" class=\"Section2\"\u003e\n \u003ch2\u003eEffects of PFPP wastewater concentration on growth and lipid production\u003c/h2\u003e\n \u003cp\u003eDue to its high ionic strength and elevated concentrations of dissolved solids, PFPP wastewater was diluted with autoclaved distilled water to obtain concentrations ranging from 0\u0026ndash;100% for the enhancement of growth and lipid production. The highest growth of \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 with a specific growth rate of 0.46\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02 day\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e and biomass productivity of 51.03\u0026thinsp;\u0026plusmn;\u0026thinsp;1.95 mg L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e day\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, was observed at 60% PFPP (Table \u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). Both lower and higher concentrations than 60% resulted in reduced biomass productivity. Similarly, the maximum lipid content and lipid productivity of 37.06\u0026thinsp;\u0026plusmn;\u0026thinsp;1.18% and 18.91\u0026thinsp;\u0026plusmn;\u0026thinsp;1.61 mg L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e day\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, respectively, was shown at 60% PFPP wastewater (Table \u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). Lipid productivity decreased at concentrations below or above 60%. These results indicate that 60% PFPP wastewater provided the most favorable nutrient composition suitable for both growth and lipid accumulation and was therefore selected for subsequent optimization experiments.\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\u0026nbsp;\u0026nbsp;\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eSpecific growth rate, doubling time, lipid content, biomass productivity, and lipid productivity of \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 cultivated under different concentrations of PFPP wastewater for 20 days. All values are presented as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation (SD) from three independent experimental replicates. Statistically significant differences among samples within the same column are indicated by different superscript letters at a 95% confidence level.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003ePFPP wastewater concentration (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003eSpecific growth rate\u003c/p\u003e\n \u003cp\u003e(day⁻\u003csup\u003e1\u003c/sup\u003e)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003eBiomass productivity\u003c/p\u003e\n \u003cp\u003e(mg L⁻\u003csup\u003e1\u003c/sup\u003e day⁻\u003csup\u003e1\u003c/sup\u003e)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003eDoubling time\u003c/p\u003e\n \u003cp\u003e(day)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003eLipid content\u003c/p\u003e\n \u003cp\u003e(%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003eLipid productivity\u003c/p\u003e\n \u003cp\u003e(mg L⁻\u003csup\u003e1\u003c/sup\u003e day⁻\u003csup\u003e1\u003c/sup\u003e)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e0.08\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e1.23\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e8.18\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e2.45\u0026thinsp;\u0026plusmn;\u0026thinsp;0.13\u003csup\u003ef\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e0.03\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e0.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e22.71\u0026thinsp;\u0026plusmn;\u0026thinsp;2.21\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e2.02\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e11.49\u0026thinsp;\u0026plusmn;\u0026thinsp;0.56\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e2.61\u0026thinsp;\u0026plusmn;\u0026thinsp;0.62\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003e40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e0.38\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e30.62\u0026thinsp;\u0026plusmn;\u0026thinsp;1.61\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e1.81\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e31.31\u0026thinsp;\u0026plusmn;\u0026thinsp;0.98\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e9.59\u0026thinsp;\u0026plusmn;\u0026thinsp;1.13\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003e60\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e0.46\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e51.03\u0026thinsp;\u0026plusmn;\u0026thinsp;1.95\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e1.52\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e37.06\u0026thinsp;\u0026plusmn;\u0026thinsp;1.18\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e18.91\u0026thinsp;\u0026plusmn;\u0026thinsp;1.61\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003e80\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e0.44\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e46.08\u0026thinsp;\u0026plusmn;\u0026thinsp;1.39\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e1.58\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e28.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.98\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e13.06\u0026thinsp;\u0026plusmn;\u0026thinsp;1.62\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e0.42\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e44.58\u0026thinsp;\u0026plusmn;\u0026thinsp;1.61\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e1.64\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e20.94\u0026thinsp;\u0026plusmn;\u0026thinsp;0.32\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e9.34\u0026thinsp;\u0026plusmn;\u0026thinsp;1.28\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec17\" class=\"Section2\"\u003e\n \u003ch2\u003eEffects of pH, temperature, and light intensity on growth and lipid production\u003c/h2\u003e\n \u003cp\u003eAt temperature of 25\u0026deg;C and a light intensity at 30 \u0026micro;mol photons m⁻\u003csup\u003e2\u003c/sup\u003e s⁻\u003csup\u003e1\u003c/sup\u003e, pH strongly influenced the growth of \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 in 60% PFPP wastewater. Increasing the pH from 4 to 8 enhanced growth and lipid production (Table \u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e).The maximum biomass productivity of 60.66\u0026thinsp;\u0026plusmn;\u0026thinsp;1.39 mg L⁻\u003csup\u003e1\u003c/sup\u003e day⁻\u003csup\u003e1\u003c/sup\u003e and lipid productivity of 28.06\u0026thinsp;\u0026plusmn;\u0026thinsp;1.60 mg L⁻\u003csup\u003e1\u003c/sup\u003e day⁻\u003csup\u003e1\u003c/sup\u003e was found at pH 8. A further increase to pH 9 resulted in a significant decline in these parameters (Table \u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). Therefore, pH 8 was considered optimal for growth and lipid production in \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01.\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\u0026nbsp;\u0026nbsp;\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eSpecific growth rate, doubling time, lipid content, biomass productivity, and lipid productivity of \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 cultivated in 60% (v/v) PFPP wastewater under different pH, temperature, and light intensities for 20 days. All values are presented as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation (SD) from three independent experimental replicates. Statistically significant differences among samples within the same column for each condition are indicated by different superscript letters at a 95% confidence level.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003epH\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003eTemperature\u003c/p\u003e\n \u003cp\u003e(\u003csup\u003eo\u003c/sup\u003eC)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003eLight intensity\u003c/p\u003e\n \u003cp\u003e(\u0026micro;mol photon m⁻\u003csup\u003e2\u003c/sup\u003e s⁻\u003csup\u003e1\u003c/sup\u003e)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003eSpecific growth rate\u003c/p\u003e\n \u003cp\u003e(day⁻\u003csup\u003e1\u003c/sup\u003e)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003eBiomass productivity\u003c/p\u003e\n \u003cp\u003e(mg L⁻\u003csup\u003e1\u003c/sup\u003e day⁻\u003csup\u003e1\u003c/sup\u003e)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003eLipid content\u003c/p\u003e\n \u003cp\u003e(%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c7\"\u003e\n \u003cp\u003eLipid productivity\u003c/p\u003e\n \u003cp\u003e(mg L⁻\u003csup\u003e1\u003c/sup\u003e day\u003csup\u003e1\u003c/sup\u003e)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\" morerows=\"5\" rowspan=\"6\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" char=\".\" colname=\"c3\" morerows=\"5\" rowspan=\"6\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.21\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e21.23\u0026thinsp;\u0026plusmn;\u0026thinsp;1.19\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e11.92\u0026thinsp;\u0026plusmn;\u0026thinsp;0.13\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c7\"\u003e\n \u003cp\u003e2.53\u0026thinsp;\u0026plusmn;\u0026thinsp;1.06\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e27.71\u0026thinsp;\u0026plusmn;\u0026thinsp;2.21\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e14.83\u0026thinsp;\u0026plusmn;\u0026thinsp;0.56\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c7\"\u003e\n \u003cp\u003e4.11\u0026thinsp;\u0026plusmn;\u0026thinsp;1.62\u003csup\u003ede\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.40\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e35.62\u0026thinsp;\u0026plusmn;\u0026thinsp;1.61\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e16.94\u0026thinsp;\u0026plusmn;\u0026thinsp;0.98\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c7\"\u003e\n \u003cp\u003e6.03\u0026thinsp;\u0026plusmn;\u0026thinsp;1.13\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.46\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e51.03\u0026thinsp;\u0026plusmn;\u0026thinsp;1.95\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e37.06\u0026thinsp;\u0026plusmn;\u0026thinsp;1.18\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c7\"\u003e\n \u003cp\u003e18.91\u0026thinsp;\u0026plusmn;\u0026thinsp;1.61\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.48\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e60.66\u0026thinsp;\u0026plusmn;\u0026thinsp;1.39\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e46.25\u0026thinsp;\u0026plusmn;\u0026thinsp;2.23\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c7\"\u003e\n \u003cp\u003e28.06\u0026thinsp;\u0026plusmn;\u0026thinsp;1.60\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.42\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e49.58\u0026thinsp;\u0026plusmn;\u0026thinsp;1.61\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e27.79\u0026thinsp;\u0026plusmn;\u0026thinsp;1.79\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c7\"\u003e\n \u003cp\u003e13.78\u0026thinsp;\u0026plusmn;\u0026thinsp;1.63\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\" morerows=\"3\" rowspan=\"4\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" char=\".\" colname=\"c3\" morerows=\"3\" rowspan=\"4\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.31\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e37.71\u0026thinsp;\u0026plusmn;\u0026thinsp;2.21\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e26.81\u0026thinsp;\u0026plusmn;\u0026thinsp;1.12\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c7\"\u003e\n \u003cp\u003e10.11\u0026thinsp;\u0026plusmn;\u0026thinsp;1.62\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.48\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e60.66\u0026thinsp;\u0026plusmn;\u0026thinsp;1.39\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e46.25\u0026thinsp;\u0026plusmn;\u0026thinsp;2.23\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c7\"\u003e\n \u003cp\u003e28.06\u0026thinsp;\u0026plusmn;\u0026thinsp;1.60\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.43\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e45.62\u0026thinsp;\u0026plusmn;\u0026thinsp;1.61\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e35.15\u0026thinsp;\u0026plusmn;\u0026thinsp;1.44\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c7\"\u003e\n \u003cp\u003e16.03\u0026thinsp;\u0026plusmn;\u0026thinsp;1.13\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\n \u003cp\u003e35\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.37\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e44.58\u0026thinsp;\u0026plusmn;\u0026thinsp;1.61\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e32.03\u0026thinsp;\u0026plusmn;\u0026thinsp;1.25\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c7\"\u003e\n \u003cp\u003e14.28\u0026thinsp;\u0026plusmn;\u0026thinsp;1.63\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" char=\".\" colname=\"c2\" morerows=\"4\" rowspan=\"5\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e1.28\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e1.95\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c7\"\u003e\n \u003cp\u003e0.03\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.48\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e60.66\u0026thinsp;\u0026plusmn;\u0026thinsp;1.39\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e46.25\u0026thinsp;\u0026plusmn;\u0026thinsp;2.23\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c7\"\u003e\n \u003cp\u003e28.06\u0026thinsp;\u0026plusmn;\u0026thinsp;1.60\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.49\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e64.62\u0026thinsp;\u0026plusmn;\u0026thinsp;1.61\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e47.18\u0026thinsp;\u0026plusmn;\u0026thinsp;1.29\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c7\"\u003e\n \u003cp\u003e30.48\u0026thinsp;\u0026plusmn;\u0026thinsp;1.63\u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.52\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e70.66\u0026thinsp;\u0026plusmn;\u0026thinsp;1.89\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e50.25\u0026thinsp;\u0026plusmn;\u0026thinsp;2.18\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c7\"\u003e\n \u003cp\u003e35.51\u0026thinsp;\u0026plusmn;\u0026thinsp;1.16\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\n \u003cp\u003e150\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.50\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e65.58\u0026thinsp;\u0026plusmn;\u0026thinsp;1.11\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e48.76\u0026thinsp;\u0026plusmn;\u0026thinsp;2.05\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c7\"\u003e\n \u003cp\u003e31.98\u0026thinsp;\u0026plusmn;\u0026thinsp;2.13\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n \u003cp\u003eTo evaluate the effect of incubation temperature, \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 was cultivated at pH 8 and a light intensity of 30 \u0026micro;mol photons m⁻\u003csup\u003e2\u003c/sup\u003e s⁻\u003csup\u003e1\u003c/sup\u003e at temperature of 20, 25, 30, and 35\u0026deg;C. Cultivation at 25\u0026deg;C yielded the highest growth, as well as the highest biomass and lipid productivities (Table \u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). Both lower (20\u0026deg;C) and higher temperatures (30\u0026ndash;35\u0026deg;C) led to reduced growth and lipid production. Therefore, 25\u0026deg;C was selected for further optimization of light intensity.\u003c/p\u003e\n \u003cp\u003eIn \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01, negligible growth and lipid production were observed under dark conditions (Table \u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). The highest growth with a specific growth rate of 0.52\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03 day\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, biomass productivity at 70.66\u0026thinsp;\u0026plusmn;\u0026thinsp;1.89 mg L⁻\u003csup\u003e1\u003c/sup\u003e day⁻1, lipid content of 50.25\u0026thinsp;\u0026plusmn;\u0026thinsp;2.18%, and lipid productivity at 35.51\u0026thinsp;\u0026plusmn;\u0026thinsp;1.16 mg L⁻\u003csup\u003e1\u003c/sup\u003e day⁻\u003csup\u003e1\u003c/sup\u003e, was achieved at a light intensity of 100 \u0026micro;mol photons m⁻\u003csup\u003e2\u003c/sup\u003e s⁻\u003csup\u003e1\u003c/sup\u003e (Table \u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). These values decreased at 150 \u0026micro;mol photons m⁻\u003csup\u003e2\u003c/sup\u003e s⁻\u003csup\u003e1\u003c/sup\u003e, likely due to the onset of light saturation. In summary, pH 8, a temperature of 25\u0026deg;C, and a light intensity of 100 \u0026micro;mol photons m⁻\u003csup\u003e2\u003c/sup\u003e s⁻\u003csup\u003e1\u003c/sup\u003e were identified as the optimal conditions for biomass and lipid production in \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 cultivated in 60% PFPP wastewater.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec18\" class=\"Section2\"\u003e\n \u003ch2\u003eGrowth and lipid production by scale-up cultivation in 2.0-L Erlenmeyer flasks\u003c/h2\u003e\n \u003cp\u003eGrowth and lipid production was further evaluated in \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 cultivated in 2.0-L Erlenmeyer flasks containing 1.8 L of PFPP wastewater under two conditions: a control conditions (100% PFPP, pH 7, 25\u0026deg;C, 30 \u0026micro;mol photons m⁻\u003csup\u003e2\u003c/sup\u003e s⁻\u003csup\u003e1\u003c/sup\u003e) and an optimal conditions (60% PFPP, pH 8, 25\u0026deg;C, 100 \u0026micro;mol photons m⁻\u003csup\u003e2\u003c/sup\u003e s⁻\u003csup\u003e1\u003c/sup\u003e). Growth by OD\u003csub\u003e750\u003c/sub\u003e measurement under the optimal conditions was approximately 1.5-fold higher than those under the control conditions (Fig. \u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eA). During 20 days of cultivation, biomass increased rapidly and reached a maximum on day 15 (Fig. \u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eB). Under the optimal conditions, \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 achieved the maximum biomass productivity of 102.79\u0026thinsp;\u0026plusmn;\u0026thinsp;3.60 mg L⁻\u003csup\u003e1\u003c/sup\u003e day⁻\u003csup\u003e1\u003c/sup\u003e, which was about 1.5 times higher than that under the control conditions (68.91\u0026thinsp;\u0026plusmn;\u0026thinsp;5.93 mg L⁻\u003csup\u003e1\u003c/sup\u003e day⁻\u003csup\u003e1\u003c/sup\u003e). Moreover, lipid production of cells cultivated under the optimal conditions was approximately 3 fold higher that under the control conditions (Fig. \u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eC). The maximum lipid content of 47.47\u0026thinsp;\u0026plusmn;\u0026thinsp;2.51% and lipid productivity of 48.80\u0026thinsp;\u0026plusmn;\u0026thinsp;3.00 mg L⁻\u003csup\u003e1\u003c/sup\u003e day⁻\u003csup\u003e1\u003c/sup\u003e was found on day 15 (Fig. \u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eC), indicating a shift from active biomass production to lipid accumulation during the mid-stationary phase.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec19\" class=\"Section2\"\u003e\n \u003ch2\u003eRemoval efficiencies of physicochemical parameters in PFPP wastewater\u003c/h2\u003e\n \u003cp\u003eThe removal of physicochemical parameters from PFPP wastewater by \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 cultivation under control and optimal conditions was evaluated. Substantial removal of all measured parameters was achieved under both conditions, with consistently higher removal efficiencies under the optimal conditions (Table \u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e). EC and COD were reduced by 85\u0026ndash;87% and 78\u0026ndash;81%, respectively, with slightly greater removal under the optimal conditions. Removal of TDS increased markedly under the optimal conditions, reaching 86%. Nitrogenous compounds were efficiently eliminated, with NH\u003csub\u003e3\u003c/sub\u003e\u0026ndash;N and NO\u003csub\u003e3\u003c/sub\u003e⁻ removal exceeding 90% under the optimal conditions. Similarly, phosphate removal improved to over 96% in the optimal conditions. These results indicate that cultivation of \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01, particularly under the optimal conditions, is highly effective for simultaneous nutrient removal and biomass/lipid production in PFPP wastewater.\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\u0026nbsp;\u0026nbsp;\u003ctable float=\"Yes\" id=\"Tab5\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eRemoval efficiencies (%) of physicochemical parameters in PFPP wastewater after cultivation of \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 under control (100% PFPP, pH 7, 25\u0026deg;C, 30 \u0026micro;mol photons m⁻\u0026sup2; s⁻\u0026sup1;) and optimal (60% PFPP, pH 8, 25\u0026deg;C, 100 \u0026micro;mol photons m⁻\u0026sup2; s⁻\u0026sup1;) conditions. Data are presented as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation (SD) (n\u0026thinsp;=\u0026thinsp;3).\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eRemoval efficiencies (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003eControl condition\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003eOptimal condition\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eEC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" char=\"±\" colname=\"c2\"\u003e\n \u003cp\u003e85.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.89\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" char=\"±\" colname=\"c3\"\u003e\n \u003cp\u003e86.55\u0026thinsp;\u0026plusmn;\u0026thinsp;0.34\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eCOD\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" char=\"±\" colname=\"c2\"\u003e\n \u003cp\u003e78.12\u0026thinsp;\u0026plusmn;\u0026thinsp;0.65\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" char=\"±\" colname=\"c3\"\u003e\n \u003cp\u003e81.46\u0026thinsp;\u0026plusmn;\u0026thinsp;0.69\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eTDS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" char=\"±\" colname=\"c2\"\u003e\n \u003cp\u003e75.81\u0026thinsp;\u0026plusmn;\u0026thinsp;2.21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" char=\"±\" colname=\"c3\"\u003e\n \u003cp\u003e86.44\u0026thinsp;\u0026plusmn;\u0026thinsp;2.12\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eNH\u003csub\u003e3\u003c/sub\u003e-N\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" char=\"±\" colname=\"c2\"\u003e\n \u003cp\u003e87.95\u0026thinsp;\u0026plusmn;\u0026thinsp;2.89\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" char=\"±\" colname=\"c3\"\u003e\n \u003cp\u003e91.06\u0026thinsp;\u0026plusmn;\u0026thinsp;1.15\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eNO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" char=\"±\" colname=\"c2\"\u003e\n \u003cp\u003e86.22\u0026thinsp;\u0026plusmn;\u0026thinsp;4.72\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" char=\"±\" colname=\"c3\"\u003e\n \u003cp\u003e95.53\u0026thinsp;\u0026plusmn;\u0026thinsp;0.46\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003ePO\u003csub\u003e4\u003c/sub\u003e\u003csup\u003e3\u0026minus;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" char=\"±\" colname=\"c2\"\u003e\n \u003cp\u003e85.54\u0026thinsp;\u0026plusmn;\u0026thinsp;2.80\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" char=\"±\" colname=\"c3\"\u003e\n \u003cp\u003e96.45\u0026thinsp;\u0026plusmn;\u0026thinsp;0.50\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n \u003cp\u003e\u003cstrong\u003eFatty acid composition and properties of biodiesel derived from lipids obtained under control and optimal conditions\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 produced lipids dominated by polyunsaturated fatty acids (PUFA) under both conditions, with a higher PUFA proportion (70.8%) under the optimal conditions compared with the control conditions (65.7%) (Table \u003cspan refid=\"Tab6\" class=\"InternalRef\"\u003e6\u003c/span\u003e). Linoleic acid (C18:2) and \u0026alpha;-linolenic acid (C18:3) were the major PUFA detected under both conditions (Table \u003cspan refid=\"Tab6\" class=\"InternalRef\"\u003e6\u003c/span\u003e). The contents of C16:2, C16:3, and eicosapentaenoic acid (EPA, C20:5) increased under the optimal conditions. In contrast, saturated fatty acids decreased from 31.4% to 25.8%, whereas monounsaturated fatty acids showed a slight increase under the optimal conditions (Table \u003cspan refid=\"Tab6\" class=\"InternalRef\"\u003e6\u003c/span\u003e).\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\u0026nbsp;\u0026nbsp;\u003ctable float=\"Yes\" id=\"Tab6\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 6\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eFatty acid composition (% of total fatty acids) of lipids obtained from \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 cells cultivated under control and optimal conditions for 20 days.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e\n \u003cp\u003eFatty acid\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e\n \u003cp\u003eFatty acid composition (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003eControl condition\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003eOptimal condition\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eMyristic acid (C14:0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e0.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003ePalmitic acid (C16:0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e27.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e23.1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003ePalmitoleic acid (C16:1)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e0.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e1.2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eHexadecadienoic acid (C16:2)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e9.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e11.8\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eHexadecatrienoic acid (C16:3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e7.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e9.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eStearic acid (C18:0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e3.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e2.2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eOleic acid (C18:1)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e2.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e2.3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eLinoleic acid (C18:2)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e26.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e26.3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003e\u0026alpha;-Linolenic acid (C18:3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e21.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e20.0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eEicosapentaenoic acid (C20:5)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e1.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e3.1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eSaturated fatty acid (SFA)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e31.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e25.8\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eMonounsaturated fatty acid (MUFA)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e2.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e3.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003ePolyunsaturated fatty acid (PUFA)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e65.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e70.7\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eTotal identified fatty acids\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n \u003cp\u003eThe biodiesel properties estimated from the fatty acid profiles are shown in Table \u003cspan refid=\"Tab7\" class=\"InternalRef\"\u003e7\u003c/span\u003e. Under the optimal conditions, several fuel-related parameters improved, including an increase in the cetane number from 54.04 to 57.79, exceeding the minimum requirements of international biodiesel standards. Cold flow properties were also enhanced, as indicated by lower cold filter plugging point and cloud point values. Although oxidation stability increased slightly, it remained below standard requirements. The saponification and iodine values decreased under the optimal conditions but remained within acceptable ranges. In contrast, the kinematic viscosity, density, and higher heating value under both conditions were lower than standard biodiesel specifications. Therefore, the shift in fatty acid composition under the optimal conditions resulted in improved biodiesel-related properties, particularly with respect to ignition quality and low-temperature performance.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eTable 7.\u003c/strong\u003e Properties of biodiesel derived from lipids obtained from \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 cells cultivated under control and optimal conditions for 20 days. Biodiesel properties were predicted using the Biodiesel Analyzer (version 1.1).\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\n \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003cimg 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\"\u003e\u003c/div\u003e\n \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003cbr\u003e\u003c/div\u003e\n \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003eDU: Degree of unsaturation (% wt.), SV: Saponification value (mg g\u003csup\u003e\u0026minus;1\u003c/sup\u003e) IV: Iodine value g I\u003csub\u003e2\u003c/sub\u003e (100 g)\u003csup\u003e\u0026minus;1\u003c/sup\u003e, CN: Cetane number, LCSF: Long-chain saturated factor, CFPP: Cold filter plugging point (\u0026deg;C), CP: Cloud point (\u0026deg;C), PP: Pour point (\u0026deg;C), APE: Allylic position equivalent, BAPE: Bis-allylic position equivalent, OS: Oxidation stability (h), HHV: Higher heating value (MJ kg\u003csup\u003e\u0026minus;1\u003c/sup\u003e), \u0026upsilon;: Kinematic viscosity (mm\u003csup\u003e2\u0026nbsp;\u003c/sup\u003es\u003csup\u003e\u0026minus;1\u003c/sup\u003e), \u0026rho;: Density (kg m\u003csup\u003e\u0026minus;3\u003c/sup\u003e)\u003c/div\u003e\n \u003c/div\u003e\n\u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eIn this study, screening of green microalgal isolates for lipid production revealed clear differences in lipid accumulation among the tested strains. Isolate SB-01 demonstrated the highest lipid content, approximately 1.2\u0026ndash;26-fold higher than that of the other isolates (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e), highlighting substantial physiological variability among green microalgae isolated from the same or different industrial wastewater sources. Such variation is common in green microalgae and is associated with differences in carbon allocation, lipid biosynthetic capacity, and stress tolerance (Nayana et al. \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). The high lipid-accumulating isolate SB-01 may be well adapted to elevated nutrient levels and capable of tolerating fluctuating environmental conditions. This adaptability likely enables SB-01 to produce biomass and accumulate lipids while simultaneously utilizing organic and inorganic compounds in wastewater for bioremediation. Comparable findings have been reported for green microalgae cultivated in sewage and laundry wastewater systems, where efficient removal of nutrients and organic loads was coupled with enhanced lipid accumulation and biodiesel-oriented biomass production (El Sheekh et al. 2023; Viena et al. 2025).\u003c/p\u003e\n\u003cp\u003eThe combined morphological and molecular analyses confirmed that isolate SB-01 belongs to the genus \u003cem\u003eCoelastrella\u003c/em\u003e. Morphological observation showed that SB-01 consisted of unicellular, bright green cells with an elliptical to slightly ovoid shape and a parietal chloroplast, which are typical characteristic of this genus. The cell size ranged from 8\u0026ndash;12 \u0026micro;m (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eA), and the cells often formed loose aggregates. These features are similar to those reported for \u003cem\u003eCoelastrella\u003c/em\u003e sp. UKM4 and \u003cem\u003eCoelastrella\u003c/em\u003e S16/8 isolated from terrestrial and wastewater environments, respectively (Ding et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Stamenov et al. \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). Molecular phylogenetic analysis based on the 18S rRNA gene further supported this identification. Isolate SB-01 was placed within a well-supported \u003cem\u003eCoelastrella\u003c/em\u003e clade that includes \u003cem\u003eC. thermophila\u003c/em\u003e, \u003cem\u003eC. aeroterrestrica\u003c/em\u003e, \u003cem\u003eC. terrestris\u003c/em\u003e, and \u003cem\u003eC. rubescens\u003c/em\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eB). The sequence showed 99.57% similarity to \u003cem\u003eC. thermophila\u003c/em\u003e var. \u003cem\u003eglobulina\u003c/em\u003e FACHB-2308. Phylogenetic analysis further placed the isolate within lineages containing known lipid- and pigment-producing \u003cem\u003eCoelastrella\u003c/em\u003e strains (Wang et al. \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Recent taxonomic revisions have also clarified species boundaries within the genus \u003cem\u003eCoelastrella\u003c/em\u003e, including the description of \u003cem\u003eC. affinis\u003c/em\u003e sp. nov. and updates within the \u003cem\u003eC. thermophila\u003c/em\u003e complex (Krivina et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). These studies highlight the high diversity within the genus, which also includes closely related taxa such as \u003cem\u003eC. multistriata\u003c/em\u003e var. \u003cem\u003emultistriata\u003c/em\u003e, \u003cem\u003eC. striolata\u003c/em\u003e var. \u003cem\u003estriolata\u003c/em\u003e, \u003cem\u003eC. corcontica\u003c/em\u003e, and members of the \u003cem\u003eC. rubescens\u003c/em\u003e lineage, together with \u003cem\u003eC. oocystiformis\u003c/em\u003e and \u003cem\u003eC. terrestris\u003c/em\u003e. Despite this diversity, many \u003cem\u003eCoelastrella\u003c/em\u003e species share similar physiological traits that allow them to adapt to environmental stress. Species of \u003cem\u003eCoelastrella\u003c/em\u003e are widely distributed in soils, aquatic systems, swamps, hot springs, and subaerial habitats, indicating strong ecological adaptability (Krivina et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). This adaptability is often linked to the accumulation of storage lipids and valuable metabolites such as carotenoids, including \u0026beta;-carotene and astaxanthin. For example, \u003cem\u003eC. striolata\u003c/em\u003e var. \u003cem\u003emultistriata\u003c/em\u003e strain 047 showed a biomass productivity of 26.67 mg L⁻\u003csup\u003e1\u003c/sup\u003e day⁻\u003csup\u003e1\u003c/sup\u003e and a lipid productivity of 6.71 mg L⁻\u003csup\u003e1\u003c/sup\u003e day⁻\u003csup\u003e1\u003c/sup\u003e under high-light conditions in humic-acid-containing medium (Susanti et al. \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Its fatty acid profile was mainly composed of oleic, \u0026alpha;-linolenic, and palmitic acids. These findings support the identification of SB-01 as a \u003cem\u003eCoelastrella\u003c/em\u003e strain and highlight its potential as a lipid-producing microalga isolated from food-processing wastewater.\u003c/p\u003e\n\u003cp\u003eThe wastewater from three sources (DPPP, FFPP, and PFPP) exhibited distinct physicochemical characteristics, reflecting differences in food-processing activities (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Such variation is commonly observed in industrial effluents and can strongly influence microalgal cultivation (Ansari et al. \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Among the tested sources, PFPP wastewater contained the highest organic and nutrient loads, as indicated by elevated COD, TDS, NH\u003csub\u003e3\u003c/sub\u003e\u0026ndash;N, NO\u003csub\u003e3\u003c/sub\u003e⁻, and PO\u003csub\u003e4\u003c/sub\u003e\u003csup\u003e3\u003c/sup\u003e⁻ concentrations (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Wastewater from processed food industries is typically rich in carbohydrates, proteins, and lipids. Consequently, it often contains high levels of organic carbon and nitrogen, which can support microalgal growth without the need for additional nutrient supplementation (Taikhao and Phunpruch \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2025\u003c/span\u003e; Ummalyma et al. \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). PFPP contained NH\u003csub\u003e3\u003c/sub\u003e-N at 21.25\u0026thinsp;\u0026plusmn;\u0026thinsp;0.56 mg L⁻\u003csup\u003e1\u003c/sup\u003e and PO\u003csub\u003e4\u003c/sub\u003e\u003csup\u003e3\u003c/sup\u003e⁻ at 4.29\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11 mg L⁻\u003csup\u003e1\u003c/sup\u003e (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). These values were higher than those measured in the other wastewater sources but substantially lower than the nutrient concentrations in the standard TAP medium (NH\u003csub\u003e3\u003c/sub\u003e-N 104.7 mg L⁻\u0026sup1; and PO\u003csub\u003e4\u003c/sub\u003e\u003csup\u003e3\u003c/sup\u003e⁻ 47.47 mg L⁻\u0026sup1;). This intermediate nutrient level likely supported active growth while imposing moderate nitrogen and phosphorus limitation. Nitrogen availability plays a key role in regulating lipid metabolism. Under nitrogen-replete conditions, microalgae typically prioritize protein synthesis and biomass accumulation. In contrast, partial nitrogen limitation redirects carbon flux toward the synthesis of neutral lipids, particularly triacylglycerols (Maltsev et al. \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Phosphorus limitation can further reduce phospholipid and nucleic acid synthesis, thereby promoting neutral lipid accumulation as an alternative carbon storage pathway (Maltsev et al. \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Therefore, the relatively lower nitrogen and phosphorus levels in PFPP compared with TAP medium may create conditions that support \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 growth while simultaneously enhancing lipid accumulation.\u003c/p\u003e\n\u003cp\u003eAlthough the nutrient-rich TAP medium produced the highest specific growth rate and biomass productivity, lipid productivity remained relatively low (Table \u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e), indicating that these conditions primarily promoted cell proliferation rather than lipid accumulation. Among wastewater sources, PFPP wastewater showed higher biomass, lipid content and lipid productivity than other sources (Table \u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). This can be attributed that PFPP wastewater contains higher NH\u003csub\u003e3\u003c/sub\u003e-N and PO\u003csub\u003e4\u003c/sub\u003e\u003csup\u003e3\u003c/sup\u003e⁻ concentrations compared with the other wastewaters, providing sufficient nutrients to support active microalgal growth. Biomass productivity in PFPP wastewater was higher than in FFPP and DPPP (Table \u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e) and exceeded values reported for \u003cem\u003eCoelastrella\u003c/em\u003e sp. and \u003cem\u003eScenedesmus dimorphus\u003c/em\u003e cultivated in municipal wastewater (Lage and Gentili \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Similar trends have been reported for other microalgae grown in organic-rich food-industry wastewaters, including \u003cem\u003eChlorella sorokiniana\u003c/em\u003e, \u003cem\u003eCoelastrella\u003c/em\u003e sp., and \u003cem\u003eChlorella pyrenoidosa\u003c/em\u003e cultivated in palm oil mill effluent (Ding et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). The lipid content obtained in PFPP wastewater (20.94%) was also substantially higher than values reported for \u003cem\u003eCoelastrella\u003c/em\u003e sp. in municipal wastewater (\u0026lt;\u0026thinsp;10%) (Lage and Gentili \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). In contrast, DPPP and FFPP wastewaters, which contain lower nutrient levels, resulted in lower growth and lipid production. This agrees with previous studies showing that dairy and low-strength food-processing wastewaters may limit nutrients or contain substances that reduce microalgal growth (Taikhao and Phunpruch \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). Interestingly, \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 was originally obtained from FFPP wastewater but exhibited significantly improved growth and lipid accumulation when cultivated in PFPP wastewater. One possible explanation is that FFPP wastewater originates from a frozen fried-food processing facility and may therefore contain relatively high levels of residual oils and lipids. Such conditions could favor the occurrence or selection of microalgae with strong lipid-accumulating capability. However, the nutrient composition of FFPP wastewater may not be optimal for sustained growth. In contrast, PFPP wastewater likely provides a more balanced nutrient environment that supports both active biomass production and lipid synthesis. These results therefore suggest that PFPP wastewater represents a suitable and low-cost cultivation medium for lipid-oriented microalgal production.\u003c/p\u003e\n\u003cp\u003eBy studying the responses of\u0026nbsp;\u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 in terms of biomass and lipid production, it was found that nutrient levels and organic load strongly influence algal growth and lipid accumulation in PFPP wastewater. The lowest growth at 0% PFPP wastewater indicates that \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 requires nitrogen and phosphorus for growth and metabolism, including photosynthesis and cell division (Table \u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). Moderate concentrations at 20\u0026ndash;40% PFPP wastewater sufficiently supported algal growth. At 60% PFPP wastewater, the highest growth rate, biomass productivity, lipid content, and lipid productivity were observed (Table \u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). This suggests that 60% PFPP wastewater provides a suitable balance between sufficient nutrients and moderate metabolic stress, which can stimulate lipid accumulation. A similar dilution-optimization concept was reported by Suh et al. (\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2024\u003c/span\u003e), where \u003cem\u003eCoelastrella\u003c/em\u003e sp. KNUA068 showed improved biomass in highly diluted cattle wastewater (1/50 diluted cattle wastewater, DCW). Likewise, \u003cem\u003eChlorella vulgaris\u003c/em\u003e and \u003cem\u003eScenedesmus\u003c/em\u003e sp. typically exhibited the highest biomass productivity at 25\u0026ndash;50% wastewater strength in industrial sewage wastewater, while higher concentrations tend to reduce biomass productivity (Singh and Rathilal, \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). At higher concentrations (80\u0026ndash;100% PFPP wastewater), growth and lipid production decreased compared with the 60% PFPP wastewater, although they remained higher than in the low-strength cultures. This reduction is likely due to excessive organic matter, high ammonium levels, and increased TDS and EC, which can cause osmotic stress, oxidative stress, and pH imbalance within the cells. Similar inhibitory effects under highly concentrated wastewater conditions have been reported for \u003cem\u003eC. striolata\u003c/em\u003e and \u003cem\u003eChlorella sorokiniana\u003c/em\u003e UKM3 cultivated in organic-rich effluents (Khalid et al. \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Susanti et al. \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Interestingly, the 60% PFPP concentration showed a growth rate similar to that of TAP medium but produced more than five times higher lipid productivity. This suggests that PFPP wastewater not only supplies essential nutrients but also provides additional organic substrates and micronutrients that are absent in defined media, while creating conditions that stimulate lipid biosynthesis in \u003cem\u003eCoelastrella\u003c/em\u003e.\u003c/p\u003e\n\u003cp\u003eThe growth and lipid productivity of \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 cultivated in PFPP wastewater were strongly influenced by pH, temperature, and light intensity. Biomass and lipid accumulation gradually increased from pH 4 to pH 8, with the maximum values observed at pH 8 (Table \u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). This indicates that the strain prefers mildly alkaline conditions. This result agrees with previous studies on many green microalgae, such as \u003cem\u003eAuxenochlorella protothecoides\u003c/em\u003e KP7, \u003cem\u003eCoelastrella\u003c/em\u003e sp., \u003cem\u003eNannochloropsis salina\u003c/em\u003e, and \u003cem\u003eTetraselmis suecica\u003c/em\u003e, where neutral to slightly alkaline pH improves inorganic carbon availability and photosynthetic efficiency (Andeden et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Singh et al. \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Arumugham et al. \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). The superior biomass and lipid production at pH 8 in \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 is also consistent with observations in \u003cem\u003eC. rubescens\u003c/em\u003e, in which slightly alkaline pH enhanced both biomass production and lipid synthesis (Minyuk et al. \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Low growth at acidic pH values (4\u0026ndash;5) is likely due to proton stress and reduced nutrient uptake. In contrast, the decrease in growth at pH 9 suggests alkaline stress and lower CO\u003csub\u003e2\u003c/sub\u003e availability, which has also been reported for \u003cem\u003eCoelastrella\u003c/em\u003e cultivated in unbuffered wastewater systems (Zhou et al. \u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Zeng et al. \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e2011\u003c/span\u003e).\u003c/p\u003e\n\u003cp\u003eTemperature clearly affected growth and lipid production, with the highest biomass and lipid productivity observed at 25\u0026deg;C. This temperature falls within the typical mesophilic range for most green microalgae, where enzyme activity and carbon fixation operate most efficiently (Farrelly et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Politaeva et al. \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Lower productivity at 20\u0026deg;C can be explained by reduced enzyme activity at low temperatures. In contrast, the decrease at high temperatures is likely due to heat stress, reduced CO₂ solubility, and damage to photosystem II (Zeng et al. \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e2011\u003c/span\u003e).\u003c/p\u003e\n\u003cp\u003eLight intensity also strongly influenced growth and lipid accumulation. Biomass and lipid accumulation increased progressively from 30 to 100 \u0026micro;mol photons m⁻\u003csup\u003e2\u003c/sup\u003e s⁻\u003csup\u003e1\u003c/sup\u003e, with a clear optimum at 100 \u0026micro;mol photons m⁻\u003csup\u003e2\u003c/sup\u003e s⁻\u003csup\u003e1\u003c/sup\u003e. This reflects the classical saturation kinetics of photosynthesis, in which increasing irradiance enhances electron transport, ATP/NADPH generation, and carbon assimilation (Souli\u0026egrave;s et al. \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). The strong enhancement of lipid accumulation at moderate irradiance (80\u0026ndash;100 \u0026micro;mol photons m⁻\u003csup\u003e2\u003c/sup\u003e s⁻\u003csup\u003e1\u003c/sup\u003e) is consistent with observations in \u003cem\u003eIsochrysis galbana\u003c/em\u003e LB987 (Gim et al. \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). However, increasing the light intensity to 150 \u0026micro;mol photons m⁻\u0026sup2; s⁻\u0026sup1; reduced productivity in \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01, likely due to photoinhibition and decreased of CO\u003csub\u003e2\u003c/sub\u003e fixation (Ashour et al. \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Similar effects have also been reported in \u003cem\u003eDunaliella salina\u003c/em\u003e and \u003cem\u003eNannochloropsis oculata\u003c/em\u003e CCAP849/1, where exposure to excessive light (200 \u0026micro;mol photons m⁻\u003csup\u003e2\u003c/sup\u003e s⁻\u003csup\u003e1\u003c/sup\u003e) reduced lipid content (Gim et al. \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2016\u003c/span\u003e).\u003c/p\u003e\n\u003cp\u003eScale-up cultivation in 2.0-L flasks confirmed that \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 can maintain strong growth and lipid production under the optimal conditions (60% PFPP wastewater, pH 8, 25\u0026deg;C, 100 \u0026micro;mol photons m⁻\u003csup\u003e2\u003c/sup\u003e s⁻\u003csup\u003e1\u003c/sup\u003e). Biomass reached a maximum of 1,542 mg L⁻\u003csup\u003e1\u003c/sup\u003e on day 15, while lipid accumulation increased later, reaching 732 mg L⁻\u003csup\u003e1\u003c/sup\u003e on day 15 (Fig. \u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). This pattern indicates a typical shift from active growth to lipid accumulation during the stationary phase. Such delayed lipid production is commonly observed in \u003cem\u003eCoelastrella\u003c/em\u003e sp. QY01 and \u003cem\u003eCoelastrella\u003c/em\u003e sp. KNUA068, where cells initially prioritize biomass formation and later redirect carbon toward lipid storage under nutrient limitation or metabolic stress (Luo et al. \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Suh et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2024\u003c/span\u003e).\u003c/p\u003e\n\u003cp\u003eComparative analysis demonstrates that \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 performs well compared with other green microalgae grown in wastewater (Table \u003cspan refid=\"Tab8\" class=\"InternalRef\"\u003e8\u003c/span\u003e). The biomass yield of 1,542 mg L⁻\u003csup\u003e1\u003c/sup\u003e and productivity of 102.79 mg L⁻\u003csup\u003e1\u003c/sup\u003e day⁻\u003csup\u003e1\u003c/sup\u003e after 15 days of cultivation were comparable to or higher than those reported for \u003cem\u003eCoelastrella\u003c/em\u003e cultivated in cattle and municipal wastewaters (Ferro et al. \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Suh et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). These values were clearly higher than those obtained in humic-rich systems and palm oil mill effluent (POME) (Ding et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Susanti et al. \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Lee et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\u0026nbsp;\u003ctable float=\"Yes\" id=\"Tab8\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 8\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eComparison of biomass yield, biomass productivity, lipid yield, and lipid productivity of \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 compared with other \u003cem\u003eCoelastrella\u003c/em\u003e strains cultivated in wastewater under different conditions.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003ccolgroup cols=\"9\"\u003e\u003c/colgroup\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eGreen microalgal species\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003eWastewater source\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003eCulture condition\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003eBiomass yield\u003c/p\u003e\n \u003cp\u003e(mg L⁻\u003csup\u003e1\u003c/sup\u003e)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003eBiomass productivity\u003c/p\u003e\n \u003cp\u003e(mg L⁻\u003csup\u003e1\u003c/sup\u003e day⁻\u003csup\u003e1\u003c/sup\u003e)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003eLipid yield\u003c/p\u003e\n \u003cp\u003e(mg L⁻\u003csup\u003e1\u003c/sup\u003e)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c7\"\u003e\n \u003cp\u003eLipid productivity\u003c/p\u003e\n \u003cp\u003e(mg L⁻\u003csup\u003e1\u003c/sup\u003e day⁻\u003csup\u003e1\u003c/sup\u003e)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c8\"\u003e\n \u003cp\u003eLipid content (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c9\"\u003e\n \u003cp\u003eReferences\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003e\u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003eProcessed food production plant wastewater\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e2.0-L Erlenmeyer flask, sterile air, 60% wastewater, pH 8, temperature at 25\u0026deg;C and light intensity of 100 \u0026micro;mol photon m⁻\u003csup\u003e2\u003c/sup\u003e s⁻\u003csup\u003e1\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e1,542\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e102.79\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e732.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c7\"\u003e\n \u003cp\u003e48.80\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c8\"\u003e\n \u003cp\u003e47.47\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c9\"\u003e\n \u003cp\u003eThis study\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003e\u003cem\u003eCoelastrella\u003c/em\u003e sp. KNUA068\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003eCattle wastewater\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e250 mL Erlenmeyer flask, shaking at 160 rpm, 100% wastewater, temperature at 25\u0026deg;C and light intensity of 7 photon m⁻\u003csup\u003e2\u003c/sup\u003e s⁻\u003csup\u003e1\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e1,470\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e147.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e333.10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c7\"\u003e\n \u003cp\u003e33.31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c8\"\u003e\n \u003cp\u003e22.66\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c9\"\u003e\n \u003cp\u003eSuh et al (\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2024\u003c/span\u003e)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003e\u003cem\u003eCoelastrella striolata\u003c/em\u003e var. \u003cem\u003emultistriata\u003c/em\u003e strain 047\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003eWater-soluble humic acid (WSHA)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e75% (v/v) WASHA at room temperature under continuous aeration and illumination at 20,000\u0026ndash;25,000\u0026nbsp;lux\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e170\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e26.67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e46.97\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c7\"\u003e\n \u003cp\u003e6.71\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c8\"\u003e\n \u003cp\u003e25.67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c9\"\u003e\n \u003cp\u003eSusanti et al (\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2024\u003c/span\u003e)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003e\u003cem\u003eCoelastrella\u003c/em\u003e sp. KKU-P1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003eUnhydrolyzed molasses\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e1.0-L PBR, ambient concentration of CO\u003csub\u003e2\u003c/sub\u003e, BBM medium supplemented with molasses (10 g L⁻\u003csup\u003e1\u003c/sup\u003e) and a sodium nitrate (1.5 g L⁻\u003csup\u003e1\u003c/sup\u003e), pH 5, temperature at 25\u0026deg;C, continuous light illumination at 23.7 W m⁻\u003csup\u003e2\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e4,280\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e305.71\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c7\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c8\"\u003e\n \u003cp\u003e15.10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c9\"\u003e\n \u003cp\u003eThepsuthammarat et al (\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2023\u003c/span\u003e)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003e\u003cem\u003eCoelastrella\u003c/em\u003e sp. KJ-04\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003eDistillery wastewater\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003eDistillery spent wash, shaking at 100\u0026nbsp;rpm, temperature at 25\u0026deg;C and light intensity of 70 \u0026micro;mol photon m⁻\u003csup\u003e2\u003c/sup\u003e s⁻\u003csup\u003e1\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e4,610\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e3,600\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c7\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c8\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c9\"\u003e\n \u003cp\u003eVasistha\u0026nbsp;et al (2023)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003e\u003cem\u003eCoelastrella\u003c/em\u003e sp.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003ePiggery wastewater\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e0.5-L Cylinder-type PBR, 5% CO\u003csub\u003e2\u003c/sub\u003e, raw piggery wastewater, temperature at 25\u0026deg;C and light intensity of 1,250 \u0026micro;mol m⁻\u003csup\u003e2\u003c/sup\u003e s⁻\u003csup\u003e1\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e7,000\u0026ndash;7,900\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e1,975\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c7\"\u003e\n \u003cp\u003e450\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c8\"\u003e\n \u003cp\u003e40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c9\"\u003e\n \u003cp\u003eLee et al (\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2021\u003c/span\u003e)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eCoelastrella sp. UKM4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003ePalm oil mill effluent (POME)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003eFlask containing 1.8 L of pre-treated POME with 1% CO\u003csub\u003e2\u003c/sub\u003e mixed with air, temperature at 25\u0026deg;C and continuous light at 20,000 lux\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e930\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e187.20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c7\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c8\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c9\"\u003e\n \u003cp\u003eDing et al (\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2020\u003c/span\u003e)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003e\u003cem\u003eCoelastrella\u003c/em\u003e sp. GN12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003eEnergy grass digestates\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003eBubbling column photobioreactors, 25% energy grass digestate: BG-11 (v: v)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e\u0026gt;\u0026thinsp;6,000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e4,940\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c7\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c8\"\u003e\n \u003cp\u003e\u0026gt;\u0026thinsp;50.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c9\"\u003e\n \u003cp\u003eXu et al (\u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2020\u003c/span\u003e)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003e\u003cem\u003eCoelastrella\u003c/em\u003e sp. (3\u0026ndash;4)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003eMunicipal wastewater\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003eMulti-Cultivator MC 1000-OD photobioreactor, municipal wastewater at 25\u0026deg;C and continuous light intensity of 100 \u0026micro;mol photon m⁻\u003csup\u003e2\u003c/sup\u003e s⁻\u003csup\u003e1\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e1,460\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e0.11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e450.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c7\"\u003e\n \u003cp\u003e34.61\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c8\"\u003e\n \u003cp\u003e30.80\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c9\"\u003e\n \u003cp\u003eFerro et al (\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2018\u003c/span\u003e)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003e\u003cem\u003eCoelastrella\u003c/em\u003e sp. QY01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003eAerobically treated swine wastewater (AnATSW)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e0.5-L Erlenmeyer flask, 40% AnATSW, 25\u0026deg;C and light intensity of 100 \u0026micro;mol photon m⁻\u003csup\u003e2\u003c/sup\u003e s⁻\u003csup\u003e1\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e625\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e57.46\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e237.50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c7\"\u003e\n \u003cp\u003e13.42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c8\"\u003e\n \u003cp\u003e24.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c9\"\u003e\n \u003cp\u003eLuo et al (\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2016\u003c/span\u003e)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eImportantly, the lipid content of 47.47%, lipid yield of 732 mg L⁻\u003csup\u003e1\u003c/sup\u003e, and lipid productivity of 48.80 mg L⁻\u003csup\u003e1\u003c/sup\u003e day⁻\u003csup\u003e1\u003c/sup\u003e after 15 days of cultivation were higher than most values reported for \u003cem\u003eCoelastrella\u003c/em\u003e grown in wastewater, including cattle, swine, municipal, and molasses-based effluents (Luo et al. \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Ferro et al. \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Thepsuthammarat et al. \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Suh et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Although some studies have reported higher lipid yields in high-strength wastewaters such as distillery effluent and digestate (Xu et al. \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Vasistha et al. \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2023\u003c/span\u003e), these systems usually contain very high organic and nitrogen loads, which can create operational challenges. In comparison, PFPP wastewater enabled \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 to achieve both high biomass productivity and high lipid accumulation under moderate-strength conditions. This balanced performance suggests that PFPP wastewater can serve as a practical and low-cost culture medium, highlighting the strong potential of \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 for large-scale, lipid-focused microalgal cultivation within circular bioeconomy systems.\u003c/p\u003e\n\u003cp\u003eThe high removal efficiencies of EC, COD, TDS, NH\u003csub\u003e3\u003c/sub\u003e\u0026ndash;N, NO\u003csub\u003e3\u003c/sub\u003e⁻, and PO\u003csub\u003e4\u003c/sub\u003e\u003csup\u003e3\u003c/sup\u003e⁻ under optimal conditions demonstrate that \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 is highly effective in treating nutrient-rich food-processing wastewater. The removal of nitrogen (91\u0026ndash;96%) and phosphate (about 96%) indicates rapid uptake and incorporation into algal biomass. This confirms the dual function of the strain as wastewater treatment and biomass production. Similarly, strains such as \u003cem\u003eCoelastrella\u003c/em\u003e sp. UKM4 and \u003cem\u003eC. terrestris\u003c/em\u003e have shown the ability to survive and actively remove nutrients from challenging waste streams such as palm oil mill effluent (Badar et al. \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Ding et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Udaiyappan et al. \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) and municipal or industrial effluents (Al Raie et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Cultivation of \u003cem\u003eCoelastrella\u003c/em\u003e sp. GN12 in energy-grass digestate also demonstrated removal efficiencies exceeding 70% for nitrogen and 90% for phosphorus, further reinforcing the robustness of this genus in nutrient-rich environments (Xu et al. \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). The strong remediation ability observed in this study is consistent with the known characteristics of the genus, such as efficient nitrogen assimilation, rapid phosphate uptake, and tolerance to high organic and salt concentrations. Improved nutrient removal under optimal pH, temperature, and light conditions also shows the close relationship between photosynthesis, biomass growth, and nutrient uptake, as previously described in carbon-rich wastewater systems (Vignesh et al. \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). These results demonstrate that \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 can achieve both high lipid production and efficient nutrient removal, making it a strong candidate for integrated wastewater treatment and value-added biomass production.\u003c/p\u003e\n\u003cp\u003eThe optimized cultivation conditions induced a significant shift in the fatty acid composition of \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01. The proportion of saturated fatty acids (SFAs) decreased from 31.5% to 25.8%, primarily due to reductions in palmitic acid (C16:0) and stearic acid (C18:0). In contrast, polyunsaturated fatty acids (PUFAs) increased from 65.6% to 70.7%, mainly driven by higher levels of C16:2, C16:3, C18:2, and C18:3, along with a more than twofold increase in eicosapentaenoic acid (EPA, C20:5). Comparable fatty acid remodeling has been reported in \u003cem\u003eCoelastrella\u003c/em\u003e sp. V3 under a two-stage cultivation strategy, where an initial growth phase was followed by a modified second stage that stimulated lipid restructuring. This strategy led to pronounced changes in the proportions of C16:0, C18:0, C18:2, and C18:3 (Minhas et al., \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). During the second stage, linoleic acid (C18:2) and \u0026alpha;-linolenic acid (C18:3) became dominant components, significantly influencing the predicted biodiesel properties. The SFA-to-PUFA shift observed in the present study highlights the metabolic flexibility of \u003cem\u003eCoelastrella\u003c/em\u003e, likely driven by enhanced desaturase activity and membrane lipid remodeling in response to the optimized cultivation conditions (Xu et al., \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2020\u003c/span\u003e).\u003c/p\u003e\n\u003cp\u003eFatty acid composition is a key determinant of biodiesel quality, particularly the balance between saturated and unsaturated fatty acids (Minhas et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Increased unsaturation generally improves cold-flow properties, whereas saturated fatty acids contribute to greater oxidative stability. In the present study, monounsaturated fatty acids (MUFAs) increased slightly from 2.92% to 3.43%. Despite this modest change, the presence of oleic acid (C18:1) remains beneficial, as this fatty acid is widely considered optimal for balancing ignition quality and oxidation resistance (Narayanan et al., \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). These compositional changes translated into improved predicted fuel properties. The cetane number increased from 54.0 to 57.8, exceeding the minimum requirements specified by the EN 14214 standard and the Thailand DOEB guidelines. In addition, the cold filter plugging point decreased from \u0026minus;\u0026thinsp;2.28\u0026deg;C to \u0026minus;\u0026thinsp;5.83\u0026deg;C, indicating improved cold-flow performance. Similar enhancements in cold-flow behavior have been reported for PUFA-rich \u003cem\u003eCoelastrella\u003c/em\u003e lipids, where increased unsaturation lowers crystallization temperature and improves low-temperature operability (Minhas et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2016\u003c/span\u003e).\u003c/p\u003e\n\u003cp\u003eTherefore, while the fatty acid profile of \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 supports acceptable ignition quality and favorable low-temperature performance, its high PUFA content may limit its direct biodiesel application as biodiesel without stabilization strategies, such as antioxidant supplementation or blending with more saturated feedstocks. At the same time, the elevated PUFA fraction particularly EPA (C20:5) and C18 polyunsaturated fatty acids enhances its potential for high-value applications, including nutraceutical and functional lipid production (Thongtha et al. \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2025a\u003c/span\u003e and \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003eb\u003c/span\u003e). Collectively, these findings highlight the dual potential of \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 for integrated wastewater-based biomass production, offering flexibility toward either biodiesel generation or PUFA-rich lipid valorization depending on downstream processing priorities and market demand.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThis study demonstrates that \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 is a robust microalgal strain capable of simultaneously achieving efficient wastewater treatment and high lipid production. Among the tested substrates, PFPP wastewater significantly enhanced biomass and lipid accumulation compared with DPPP and FFPP. Under optimized conditions (60% PFPP, pH 8, 25\u0026deg;C, and 100 \u0026micro;mol photons m⁻\u0026sup2; s⁻\u0026sup1;), the strain achieved a biomass productivity of 102.79 mg L⁻\u0026sup1; day⁻\u0026sup1;, lipid content of 47.47%, and lipid productivity of 48.80 mg L⁻\u0026sup1; day⁻\u0026sup1;, while removing 81\u0026ndash;96% of nutrients and organic pollutants. The PUFA-rich fatty acid profile further indicates potential for both biodiesel production and high-value lipid applications. Therefore, \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 shows strong promise for integrated biorefinery systems that convert agro-industrial wastewater into value-added biofuels and functional lipids, supporting circular bioeconomy strategies.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors appreciation to the three food industry factories in Phra Nakhon Si Ayutthaya Province for their kind cooperation in providing wastewater samples for this study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors’ contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eConceptualization, S.T. and S.P.; methodology, S.T.; software, S.T.; validation, S.T. and S.P.; formal analysis, S.T. and S.P.; investigation, S.T.; resources, S.P.; data curation, S.T.; writing—original draft preparation, S.T. and S.P.; writing—review and editing, S.T. and S.P.; visualization, S.T.; supervision, S.P.; project administration, S.T., and S.P.; funding acquisition, S.T. and S.P. All authors have read and agreed to the published version of the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research did not receive funding.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAl Raie HH, Al Hassany JS, Rasheed KA (2020) Wastewater treatment using \u003cem\u003eCoelastrella terrestris\u003c/em\u003e. Plant Arch 20:1691\u0026ndash;1695.\u003c/li\u003e\n\u003cli\u003eAmerican Public Health Association (2017) Standard Methods for the Examination of Water and Wastewater, 23\u003csup\u003erd\u003c/sup\u003e edn. APHA, Washington DC.\u003c/li\u003e\n\u003cli\u003eAndeden EE, Ozturk S, Aslim B (2021) Effect of alkaline pH and nitrogen starvation on the triacylglycerol (TAG) content, growth, biochemical composition, and fatty acid profile of \u003cem\u003eAuxenochlorella protothecoides\u003c/em\u003e KP7. 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Environ Eng Res 19:283\u0026ndash;287.\u003c/li\u003e\n\u003cli\u003eXu J, Zhu S, Mo N, Wang Z (2020) Screening of freshwater oleaginous microalgae from South China and its cultivation characteristics in energy grass digestate. J Clean Prod 276:124193.\u003c/li\u003e\n\u003cli\u003eYeesang J (2018) Screening and cultivation of oleaginous microalgae in industrial wastewater: possible applications for wastewater treatment and feedstock production. Interdiscip Res Rev 12:48\u0026ndash;56.\u003c/li\u003e\n\u003cli\u003eZeng X, Danquah MK, Chen XD, Lu Y (2011) Microalgae bioengineering: from CO\u003csub\u003e2\u003c/sub\u003e fixation to biofuel production. Renew Sustain Energy Rev 15:3252\u0026ndash;3260.\u003c/li\u003e\n\u003cli\u003eZhou W, Wang J, Chen P et al (2017) Bio-mitigation of carbon dioxide using microalgal systems. Renew Sustain Energy Rev 76:1163\u0026ndash;1175.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"","identity":"journal-of-applied-phycology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"","snPcode":"10811","submissionUrl":"https://submission.nature.com/new-submission/10811/3","title":"Journal of Applied Phycology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Coelastrella, Wastewater, Biomass, Lipid, Biodiesel","lastPublishedDoi":"10.21203/rs.3.rs-9112207/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-9112207/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eCultivating microalgae in wastewater represents a sustainable strategy for generating lipid-rich biomass for biodiesel production while concurrently achieving wastewater remediation. In this study, green microalgae isolated from food industry wastewater were screened to identify strains with high lipid production potential. The most promising isolate was identified by morphological characteristics and 18S rRNA gene analysis as \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01. Comparative cultivation in three types of food industry wastewater showed that wastewater from a processed food production plant (PFPP) supported the highest biomass growth and lipid production. Optimization of wastewater concentration indicated that 60% (v/v) PFPP wastewater was optimal for biomass and lipid production. Further optimization demonstrated that pH 8, 25\u0026deg;C, and a light intensity of 100 \u0026micro;mol photons m⁻\u003csup\u003e2\u003c/sup\u003e s⁻\u003csup\u003e1\u003c/sup\u003e maximized growth, yielding biomass and lipid productivities of 70.66\u0026thinsp;\u0026plusmn;\u0026thinsp;1.89 and 35.51\u0026thinsp;\u0026plusmn;\u0026thinsp;1.16 mg L⁻\u003csup\u003e1\u003c/sup\u003e day⁻\u003csup\u003e1\u003c/sup\u003e, respectively. Scale-up cultivation in 2.0-L flasks further increased biomass productivity to 102.79\u0026thinsp;\u0026plusmn;\u0026thinsp;3.60 mg L⁻\u003csup\u003e1\u003c/sup\u003e day⁻\u003csup\u003e1\u003c/sup\u003e, with a lipid productivity of 48.80\u0026thinsp;\u0026plusmn;\u0026thinsp;3.00 mg L⁻\u003csup\u003e1\u003c/sup\u003e day⁻\u003csup\u003e1\u003c/sup\u003e, and lipid content of 47.47\u0026thinsp;\u0026plusmn;\u0026thinsp;2.51%. Simultaneously, nutrient and organic pollutant removal efficiencies reached 81\u0026ndash;96% for COD, TDS, NH\u003csub\u003e3\u003c/sub\u003e\u0026ndash;N, NO\u003csub\u003e3\u003c/sub\u003e⁻, and PO\u003csub\u003e4\u003c/sub\u003e\u003csup\u003e3\u003c/sup\u003e⁻. The resulting PUFA-rich fatty acid profile supports both biodiesel production and high-value PUFA applications, highlighting \u003cem\u003eCoelastrella\u003c/em\u003e sp. RMUTSB 01 as a promising candidate for integrated wastewater-based lipid production within a sustainable biorefinery framework.\u003c/p\u003e","manuscriptTitle":"Integrated screening and cultivation optimization of food processing wastewater-derived microalgae for enhanced biomass and lipid production toward sustainable biodiesel","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-03-31 12:12:56","doi":"10.21203/rs.3.rs-9112207/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-04-27T21:36:40+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-04-21T02:57:35+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-04-18T10:08:08+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"333815749673539922559327123912854698045","date":"2026-04-01T09:44:25+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"208420396526074198810642984990345734624","date":"2026-03-30T05:07:20+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"261132208584151769432818770070158506676","date":"2026-03-28T18:01:25+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"214571574407004724022270688977207451074","date":"2026-03-28T01:32:12+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2026-03-27T08:18:37+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2026-03-27T06:47:45+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2026-03-24T10:11:43+00:00","index":"","fulltext":""},{"type":"submitted","content":"Journal of Applied Phycology","date":"2026-03-13T08:08:17+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"","identity":"journal-of-applied-phycology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"","snPcode":"10811","submissionUrl":"https://submission.nature.com/new-submission/10811/3","title":"Journal of Applied Phycology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"1822847d-a330-4f0b-9acd-c2e510d850e4","owner":[],"postedDate":"March 31st, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2026-05-14T14:08:22+00:00","versionOfRecord":[],"versionCreatedAt":"2026-03-31 12:12:56","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-9112207","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-9112207","identity":"rs-9112207","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}