Rice-Based Distillers’ Dried Grains from Bio-Ethanol Production, a Potential Source of Protein for Food Application

Rice-Based Distillers’ Dried Grains from Bio-Ethanol Production, a Potential Source of Protein for Food Application | 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 Rice-Based Distillers’ Dried Grains from Bio-Ethanol Production, a Potential Source of Protein for Food Application Tien-Thanh Nguyen, Gia Long Nguyen, Bach Cao-Xuan, Thuy Nguyen-Thanh, and 5 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3171967/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Several bioethanol plants in Vietnam using rice or cassava as raw materialsgenerate a large number of by-products in the form of wet distillers spent grain, which have been underutilized with simple treatment for animal feeding. In this work, the biochemical and nutritional values such as protein (amino acid profile), lipid, fiber, ash, starch, calcium and phosphorous of dried distillers’ grain (DDG) collected from different bioethanolfactories in Vietnam were assessed. The DDG samples were shown to vary in nutrient compositions which depended either on raw materials or ethanol processing technology. Among, rice DDG was shown as the most nutritively valuable with very high protein content (55-80% of dry matter) and appropriate amino acid profile whereas cassava DDG was characterized by a high fiber content and a low protein content (13-16%). The protein in rice based DDG could be enriched by extraction/precipitation and applied for food products. The obtained data suggest that the by-products from rice-based bioethanol are very potential and promise to be used efficiently as ingredients not only for the animal feed industry but also for the food industry. rice cassava bioethanol by-product proximates non-animal protein food application Figures Figure 1 Figure 2 Figure 3 Highlights The first systematic analysis of composition of DDG from rice and cassava- based bioethanol factory show the variation of proximates depending on technologies and raw materials Rice DDG is high potential of non-animal protein for food application. Cassava DDG can be valorized by solid state fermentation with thermostable fungi to improve protein content and accumulate beneficial enzymes for animal digestion system 1. Introduction Distiller’s dried grain with soluble (DDGS) has been known as a co-product of the bioethanol industry from corn or wheat [ 1 ]. The composition of DDGS has been received a great interest in animal science, ethanol producers, and especially in the animal feeding industry as most of this co-product has been sold as feed ingredients for livestock [ 2 ]. DDGS was also considered as a potential source of insoluble dietary fiber which could be beneficial in the prevention of colon cancer, coronary heart disease, diabetes, and obesity. Therefore, DDGS was recently valorized as an ingredient for food application [ 3 – 7 ]. Bioethanol including for biofuel and beverage production is highly demanded in Vietnam [ 8 ]. The expected production of bioethanol in Vietnam is about 170 million liters for the food industry by 2035 [ 8 ]. There are several bioethanol factories using different production technologies with rice or cassava as starchy materials. Regardless of raw materials, it is estimated that to gain 1 liter of ethanol, 2.3 kg of material are used and about 0.15 kg of dry matter (6.5% of raw material) of distiller grain are produced. This means that by 2035, approximately 0.65 million tons of dried distiller grain will be generated in Vietnam. The wet distiller grain (WDG) and dried distiller grain (DDG) from ethanol factories has been used since a long time ago for the purpose of feeding in Vietnam. These by-products usually are directly added to the diet of animals without or with simple treatment [ 9 ]. Few studies have started to focus on cassava and rice DDG, mainly targeting animal feeding. For example, authors used rice and DDG in pig feed and showed beneficial effects on animal performance [ 10 ]. Cassava DDG has been improved the protein content by solid fermentation with yeasts or molds [ 11 ]. However, the potential of these DDGs, especially rice-DDG could be extended because rice is used as the staple food in Vietnam and many Asian countries. This work aimed to elucidate systematically the nutrient composition of DDG samples from different bioethanol factories in Vietnam which use different technologies on either cassava or rice as raw materials. Furthermore, the protein content of DDG, especially rice based DDG was preliminarily improved and applied in food products. The results showed the high potential of DDG from rice-based bioethanol in the food industry. 2. Materials and methods 2.1 Samples collection Seven wet distillers spent (WDS) samples were collected from 5 ethanol factories located over the country (Table 1 ). The WDS samples were transported immediately after production directly to Hanoi University of Science and Technology (Vietnam) and were dried following a protocol as follows: 90 o C for 30 min, 80 o C for 2.5-3 h and finally 70 o C for 1 h in a circulating dryer. The dried distiller grain (DDG) samples then were packed in plastic bags and stored at -10 o C for further analysis. Depending on the raw material for bioethanol production, the samples were noted with R (rice), CS (cassava) or C (corn). Another sample (named SLSF) was obtained from a simultaneous liquefaction, saccharification and fermentation process (SLSF) conducted at factory 2 in the Northern with rice (Table 1 , Fig. 1 d). Table 1 Overview of ethanol production and wet distillers’ grain (WDG) in 5 factories in Vietnam Samples SLSF Factory 1 Factory 2 Factory 3 Factory 4 Factory 5-CS Factory 5-CO Capacity (L/year) 150,000 1,500,000 1,200,000 10,000,000 100,000,000 76,000,000 76,000,000 End user Beverage ethanol Beverage ethanol Beverage ethanol Beverage ethanol Fuel ethanol Fuel ethanol Beverage ethanol Raw materials Rice Rice Rice Rice Cassava Cassava Corn Processing Simultaneous liquefaction, saccharification and fermentation at pilot scale (500 L/batch) Separate liquefaction, saccharification and fermentation Separate liquefaction, saccharification and fermentation Separated Liquefaction Simultaneous Saccharification and fermentation Separated Liquefaction Simultaneous Saccharification and fermentation Liquefaction, saccharification and continuous fermentation Liquefaction, saccharification and continuous fermentation Separation Wet distillers’ grains (WDG) were filtered before distillation by a frame and plate filtration WDG were filtered before distillation by a frame and plate filtration WDG were filtered before distillation by a frame and plate filtration WDG were separated after distillation by decanter WDG were separated after distillation by decanter WDG were filtered after distillation by a frame and plate filtration WDG were filtered after distillation by a frame and plate filtration Drying No No No No Flash drying Flash drying Flash drying 2.2. Proximate composition analysis Proximate composition in DDG samples was analyzed by using the standard methods including lipid (ISO 6492:1999), ash (ISO 05984:2002), crude fiber (with Ankom filter bag technique), total crude protein (ISO 05983-1:2005), phosphorus (ISO 6491:1998), calcium (ISO 06490:1985). Starch was determined via reducing sugar generated by hydrolyzing samples in HCl 2% for 2 hours in a boiling water bath. The reducing sugar content was determined by using dinitrosalicylic acid [ 12 ]. 2.3 Amino acid profile determination The amino acid profile in DDG samples was determined as previously described. Shortly, samples (ca. 40 mg) were hydrolyzed with vapor of HCl 6M, 0.5% phenol for 24h at 120 o C. The amino acids in the neutralized hydrolysates were derivatized with OPA, separated on the column C18 ElipseZorbax 5 µm, 4.6 x 150 mm (Agilent, US) and detected by DAD detector of Agilent 1200 series HPLC (USA). Two mobile phases, sodium phosphate 40 mM, pH 7.8 and methanol: acetonitrile: deionized water = 45:45:10, were used in the 31-minute gradient elution program as previously described [ 10 , 11 ]. 2.4. Preliminary valorization of rice DDG and cassava DDG Extraction and application of protein in rice SLSF DDG Protein in rice-SLSF DDG was extracted by using the alkaline solution as described previously [ 13 ]. The rice DDG from SLSF was mixed at the ratio of 1:30 weight/volume with NaOH 0.5M and NaHSO 3 50 mM. The mixtures were stirred at 600 rpm and maintained at different temperatures of 25 o C, 50 o C and 70 o C with a water bath (Memmert WNB 22, Germany) for 2 hours. Mixtures then were centrifuged at 6000 ×g/15 min to separate supernatants and residue solids. The residue solids were washed with water, dried, and determined the dry weight and the protein content. The supernatants were adjusted pH to 4.0 with HCl to precipitate the extracted protein. Precipitates were collected by a similar centrifuging step. Similarly, the precipitates were washed with water, dried, and used for measurement of protein. The precipitate which was enriched in protein content then was preliminarily applied to bread recipe by replacement of wheat flour. The ratio of replacement was from 5–20%. Bread quality parameters including rheological/sensorial parameters such as hardness, cohesiveness, resilience, chewiness, springiness, gumminess were measured by using a TA.XT – plus texture analyzer (Stable Micro Systems, UK) with 5 kg loadcell sensor, 25mm diameter probe (P/25). The texture profile of the bread was measured 1 hour after baking. The breads were sliced at the largest circumference with slice thickness of 20 mm. The speed was of 5mm/s and the compressive deformation was 50% (10mm). Solid-state cultivation of cassava DDG by thermophilic fungi Cassava DDG (Factory 5-CS) was used for solid-state fermentation with four thermophilic fungal strains including Rhizomucor miehei FCH 116.3; Rhizopus microsporus LPH 084; Rhizopus microsporus LPH 156; Thermothelomyces thermophilus FCH 112.2 [ 14 ]. Fungal spores on PDA plates were collected after 5 days of cultivation at 50°C and suspended in 4 mL of sterile saline 0.9% w/v and 0.05% Tween 80 and used as inoculums. Two milliliters of each inoculum were added into cassava WDG (~ 5 g dry matter) in a 500 mL Erlenmeyer flask. After 7 days of incubation at 50°C, the protein content and enzyme activities (xylanase, CMCase, amylase) in solid phases were measured. For protein measurement, the dried solid was used as described above. For enzyme extraction, 50 mM sodium citrate buffer pH 5.0 was added to the solid medium in flasks and mixed for 1 h at 30°C at 160 rpm. The supernatant was obtained by centrifugation at 6000 ×g/10 min at 10°C and stored at − 20°C until analysis. For the determination of xylanase, CMCase and amylase activity, the extracts were filtrated through 0.45 µm filter (Minisart® NY25 Syringe Filter) and used for enzyme reaction with substrates including 1% xylan (xylan from beechwood, Apollo Scientific®), CMC (Sigma-Aldrich) and Starch (Samchun chemicals®) in 50 mM sodium citrate pH 5 buffer, respectively. The enzyme reactions were performed at 50 o C for 20 min including 0.1 ml of extract and 0.2 ml of substrate solution. The enzymatically generated reducing sugar concentrations were measured by using dinitrosalicylic acid (DNS) as mentioned above. 2.6 Data analysis Descriptive statistical parameters such as mean values and standard errors were calculated using Microsoft Excel from at least duplicate measurements. 3. Results Seven WDS samples were collected from 5 factories that are different in raw materials, capacity, ethanol production process and WDG harvesting and treatment (Table 1 , Fig. 1 ). Among, 4 samples, named SLSF-R, Factory 1-R, Factory 2-R and Factory 3-R, were from rice material (rice-based); 2 samples Factory 4-CS and Factory 5-CS were from cassava material (cassava-based); and the rest (Factory 5-CO) was based on corn material. Seven DDG samples were obtained after drying, respectively. These DDG samples were analyzed for the proximate components and amino acid profiles. 3.1. Proximate composition of rice-based and cassava DDG samples DDG samples obtained from 5 factories were different in proximate composition as shown in Table 2 . Table 2 Composition of Dried Distillers Grain (DDG) samples Compositions SLSF-R Factory 1-R Factory 2-R Factory 3-R Factory 4-CS Factory 5-CS Factory 5-CO Ref DDGS Raw materials Rice Rice Rice Rice Cassava Cassava Corn Corn Crude Protein (% DM) 51.49 ± 0.84 72.44 ± 1.36 74.96 ± 2.40 79.60 ± 1.13 13.38 ± 0.64 16.40 ± 0.72 35.60 ± 0.82 31.40 Non-protein (% DM) Starch 15.01 ± 0.17 14.29 ± 0.57 11.77 ± 0.51 8.55 ± 0.38 19.43 ± 0.86 10.90 ± 0.25 26.63 ± 1.02 5.30 Crude Fiber 18.42 ± 0.82 9.09 ± 0.27 11.58 ± 0.42 8.98 ± 0.14 46.72 ± 1.70 36.50 ± 1.70 7.59 ± 0.21 10.20 Fats 7.47 ± 0.25 10.41 ± 0.42 2.11 ± 0.07 0.69 ± 0.03 2.30 ± 0.07 2.94 ± 0.11 4.56 ± 0.11 12.00 Ash 1.74 ± 0.07 2.27 ± 0.10 1.44 ± 0.06 2.61 ± 0.10 9.05 ± 0.27 12.08 ± 0.42 3.00 ± 0.14 4.60 Total non-protein (%) 42.64 36.06 26.90 20.83 77.50 62.42 41.78 32.10 Calcium (% DM) 0.11 0.02 0.04 0.14 0.41 - - - Phosphorus (% DM) 0.02 0.02 0.03 0.03 0.01 - - - R: rice; CS: cassava; CO: corn The protein in rice DDG amounted to 50–80% of dry matter, while in cassava DDG, this value was only 13.38–16.40% of dry matter. The protein composition in the corn DDG Factory 5-CO was 35.6% of dry matter. The starch content was measured in a range of 8.55–15.01% of dry matter for the rice DDG, and 10.90-19.43% for the cassava DDG. Meanwhile, the starch contributed up to 26.63% of dry matter in the corn DDG sample. The fiber content was found inconsistent among these DDGs. The cassava DDG (Factory 4-CS and Factory 5-CS) contained significantly high fiber, which was 46.72% for the former and 36.5% for the latter. The fiber content of the rice DDGs varied in a range of 9.00–11.58%, except for the SLSF-R which was 18.42%. The corn DDG Factory 5-CO contained a slightly lower fiber compared to the rice DDG, amounting to 7.59% of dry matter. Fat content was found to rather varied from sample to sample. Fat was at a higher level in the rice DDG such as SLSF-R (7.47%), Factory 1-R (10.41%). Surprisingly, the DDG Factory 3-R contained rather low fat (0.69%). The fat in cassava DDGs varied in a narrow range of 2.30–2.94% of dry matter. The fat content in corn DDG Factory 5-CO was 4.56% of dry matter The ash of rice DDG samples amounted to 1.44–2.61% of dry matter. This component was slightly higher in the corn DDG (3%). In contrast, ash contributed a high level in the cassava DDG samples ranging from 9.05 to 12.08%. Calcium and phosphorus were also figured out for the rice DDGs and one cassava DDG sample. Phosphorus was at a low level in all the test samples (between 0.01–0.03% of dry matter). Calcium was very low in the rice DDG Factory 1-R (0.02%), but much higher in the cassava DDG Factory 4-CS. 3.2. Amino acid profiles Table 3 shows the consistent results on amino acid profiles. The amino acid profile was generally similar among the DDG samples which derived from the same type of raw material. In agreement with the protein composition, the amino acid was lower in cassava and corn DDG than in rice DDG samples. Most of the essential amino acids with significant amounts were determined in the DDG, especially the rice DDG samples. Leucine, phenylalanine, arginine, lysine contributed more than 3% of dry matter in the rice DDGs. These amino acids amounted much lower in cassava and corn DDGs, except leucine in the corn DDG Factory 5-CO sample which was comparable to that of rice DDG (Table 3 ). Table 3 Profile of amino acids of DDG samples Amino acids (%) SLSF-R Factory 1-R Factory 2-R Factory 3-R Factory 4-CS Factory 5- CS Factory 5-CO Essentials amino acids HIS 1.05 ± 0.02 1.65 ± 0.02 1.35 ± 0.02 1.64 ± 0.04 0.31 ± 0.04 0.27 ± 0.02 0.93 ± 0.02 ARG 2.60 ± 0.06 3.41 ± 0.11 4.44 ± 0.02 3.44 ± 0.04 1.22 ± 0.04 1.21 ± 0.03 1.91 ± 0.03 THR 1.87 ± 0.07 2.17 ± 0.05 2.23 ± 0.03 2.28 ± 0.04 0.50 ± 0.03 0.70 ± 0.06 0.99 ± 0.01 VAL + MET 2.17 ± 0.08 2.52 ± 0.06 2.79 ± 0.06 3.26 ± 0.05 0.41 ± 0.02 0.68 ± 0.02 0.95 ± 0.02 PHE 2.57 ± 0.08 3.38 ± 0.09 3.41 ± 0.05 3.48 ± 0.06 0.61 ± 0.05 1.12 ± 0.02 1.70 ± 0.04 ISOLEU 2.22 ± 0.06 2.26 ± 0.11 2.39 ± 0.02 2.79 ± 0.03 0.50 ± 0.03 0.89 ± 0.01 1.07 ± 0.01 LEU 3.86 ± 0.05 4.48 ± 0.05 4.85 ± 0.02 4.68 ± 0.01 0.87 ± 0.05 1.45 ± 0.02 4.61 ± 0.10 LYS 2.53 ± 0.03 3.66 ± 0.13 3.59 ± 0.04 4.16 ± 0.03 0.90 ± 0.02 0.99 ± 0.02 0.99 ± 0.01 Non-essential amino acids GLY 0.54 ± 0.03 1.54 ± 0.17 1.49 ± 0.03 1.83 ± 0.03 0.11 ± 0.01 0.14 ± 0.01 0.37 ± 0.02 ASP 4.17 ± 0.06 8.25 ± 0.19 7.88 ± 0.11 7.70 ± 0.01 2.39 ± 0.07 2.44 ± 0.03 3.70 ± 0.09 GLU 7.37 ± 0.02 8.49 ± 0.05 9.05 ± 0.05 9.05 ± 0.03 1.47 ± 0.06 2.26 ± 0.06 6.10 ± 0.06 SER 1.95 ± 0.06 2.41 ± 0.03 2.38 ± 0.01 2.48 ± 0.06 0.65 ± 0.04 0.73 ± 0.05 1.38 ± 0.02 ALA 3.14 ± 0.07 3.33 ± 0.03 3.67 ± 0.02 3.21 ± 0.03 1.11 ± 0.09 1.54 ± 0.02 3.24 ± 0.03 TYR 1.50 ± 0.05 2.62 ± 0.07 2.51 ± 0.01 3.09 ± 0.06 0.19 ± 0.02 0.25 ± 0.02 1.20 ± 0.06 CYS 1.11 ± 0.07 2.09 ± 0.08 2.23 ± 0.02 3.25 ± 0.04 0.21 ± 0.03 0.34 ± 0.02 0.57 ± 0.02 The values were the averages of duplicate experiments. DDG samples also contained most of the non-essential amino acids. Aspartic acid and glutamic acid were found as the highest amino acids in most of the DDGs regardless of the raw material. These amino acids contributed up to 7.7–9.0% of dry matter in the rice DDGs (except the SLSL-R, due to the lower protein content of this sample compared to other rice DDGs) and 3.7–6.1% of dry matter in the corn DDG Factory 5-CO. Alanine was found also in these DDGs at a remarkable level, especially in the rice and corn DDG, in which alanine contributed more than 3% of dry matter (Table 3 ). Amino acids of cassava DDG samples were lower in accordance with the lower protein ratio. Arginine, leucine, aspartic acid, glutamic acid, and alanine presented in cassava DDGs at the level above 1%. In contrast, histidine, glycine, tyrosine and cysteine amounted below the level of 0.34% (Table 3 ). 3.3. Preliminarily valorization of rice and cassava DDG At different temperatures, NaOH 0.5M (with NaHSO 3 0.05M) extracted a varied amount of solid from SLSF-R. For instance, at 70 o C in 2 hours, 79.31% dry weight of DDG was extracted. At lower temperatures such as 50 o C and 25 o C, the extraction yield was reduced to 65.66 and 56.44%, respectively (Table 4 ). By adjusting pH to 4, the precipitation was generated from the extracted soluble in liquid phase. However, the precipitate amount was not in agreement with the extraction yields. For instance, the precipitate from 50 o C extraction was almost the same as that from 70 o C extraction (ca. 36% of the initial DDG amount). The extraction at 25 o C resulted in less extracted yield and precipitation (about 27.14% of the initial DDG amount) after pH adjustment (Table 4 ). The protein content in precipitates was in the range of 59.72 to 72.5% (Table 4 ) which was improved than the protein content in the initial SLSF-R (51.49%) (Table 2 ). Table 4 Extraction of protein from SLSF-R Condition of extraction Extraction yield** (%) Residual solid* (%) Precipitated solid* (%) Protein content in precipitated solid (%) Protein recovery (%) 70℃, 120 rpm, 2 hours 79.31 ± 1.47 20.69 36.22 ± 1.53 70.53 ± 2.23 54.82 50℃, 120 rpm, 2 hours 65.66 ± 1.37 34.34 35.90 ± 0.31 72.50 ± 2.57 55.85 25℃, 120 rpm, 2 hours 56.44 ± 2.45 43.56 27.14 ± 1.82 59.72 ± 1.05 34.78 * weight of solid in comparison to the initial SLS-R amount; ** calculated based on the residual solid The bread samples with the replacement of wheat flour by enriched protein precipitate showed differences in rheological properties compared to the control bread (without replacement) (Table 5 , Fig. 2 ). In general, the more replacement, the higher hardness, chewiness, and gumminess of bread samples was observed (Table 5 ). Table 5 Rheological property of bread samples with different wheat replacement ratio by enriched protein Parameters Ratio of enriched protein 0% 5% 10% 15% 20% Hardness(g) 109.69 ± 11.44 336.40 ± 32.59 669.16 ± 77.30 1037.06 ± 80.39 1496.50 ± 94.39 Cohesiveness 0.908 ± 0.013 0.85 ± 0.01 0.764 ± 0.02 0.660 ± 0.034 0.597 ± 0.033 Resilience 0.614 ± 0.072 0.849 ± 0.092 0.433 ± 0.022 0.328 ± 0.032 0.298 ± 0.062 Chewiness (N) 98.67 ± 15.01 263.48 ± 28.42 420.29 ± 36.14 612.124 ± 33.33 727.00 ± 53.08 Springiness 0.918 ± 0.011 0.949 ± 0.017 0.922 ± 0.023 0.871 ± 0.026 0.859 ± 0.022 Gumminess 113.70 ± 12.90 265.67 ± 15.84 451.36 ± 22.50 703.87 ± 49.53 845.91 ± 57.02 Four fungal strains were used in the solid-state cultivation of cassava DDG Factory 5-CS. The protein content after 7 days of cultivation was slightly improved, ranging from 18.08–20.70%, from the initial protein value in Factory 5-CS was 16.42% (Table 6 ). On the other hand, the measure of three carbohydrate-active enzymes including CMCase, xylanase and amylase showed that they were produced and accumulated in solid media at varied activity levels (Table 6 , Fig. 3 ). Table 6 Changes of proximate composition and enzyme accumulation of Factory 5-CS in solid state cultivation with fungal strains Strain Fiber (%DM) Ash (%DM) Protein (%DM) CMCase (U/g DM) Xylanase (U/g DM) Amylase (U/g DM) Factory 5-CS* 33.98 ± 1.28 10.03 ± 0.59 16.42 ± 0.16 0 0 0 Rhizomucor miehei FCH 116.3 35.55 ± 0.68 18.67 ± 0.43 19.73 ± 0.44 0.33 ± 0.07 0.5 ± 0.05 10.07 ± 0.34 Rhizopus microsporus LPH 084 35.76 ± 0.90 12.59 ± 0.93 18.58 ± 0.21 0.66 ± 0.39 0.93 ± 0.35 14.25 ± 0.73 Rhizopus microsporus LPH 156 37.41 ± 1.45 12.22 ± 0.71 18.08 ± 0.16 1.02 ± 0.76 1.39 ± 0.67 14.44 ± 0.84 Thermothelomyces thermophilus FCH 112.2 36.35 ± 0.66 15.17 ± 0.60 20.70 ± 0.12 1.00 ± 0.22 4.94 ± 0.42 2.03 ± 0.63 * Sample collected in a different batch from the sample of cassava DDG in Table 2 4. Discussion The wet by-product from the ethanol industry has been used for a long time for the purpose of animal feeding in Vietnam. However, the nutrient composition of this potential by-product has not been elucidated, except for little data recently published [ 10 , 11 ]. In contrast, there is a huge bank of data for wheat and corn DDGS which have been published several years [ 2 , 15 – 17 ]. In fact, rice and cassava have been known as more popular crops in Asian and African countries than maize and wheat. This study was the first systematic work focusing on the nutrient compositions of by-products from different ethanol factories in Vietnam using rice, and cassava as raw materials. 4.1 The composition of DDG depend on bioethanol technology and raw materials In this study, the variation of composition including crude protein, starch, fat, crude fiber and ash of 7 DDG samples collected from 5 ethanol factories was observed. The variation in composition was also proven for DDGS from corn and wheat [ 2 , 16 ]. The authors showed that the types of raw material, the production technology and WDG treatment influence on the composition of DDGS, and this was an explanation for the variation in proximates observed with rice and cassava DDGs in this study. Proximate of these DDGs were different due to not only the raw material but also ethanol processing and WDG treatment (Table 1 , Fig. 1 ). In general, for all materials, bioethanol manufacture starts with a liquefaction step in which milled raw material is treated at high temperature of above 100 o C and supported by a heat-stable α-amylase. Among these, a high-pressure steam treatment (temperature of 105 -110 o C) was applied in Factory 3 and Factory 5 to achieve the highly extensive liquefaction of starch. A saccharification step was performed either separately (as in Factory 1, Factory 2 and Factory 5) or simultaneously with fermentation (as in Factory 4 and Factory 5). Factory 5 carried out a continuous fermentation meanwhile others used batch fermentation. The SLSF sample was different from others because the liquefaction, saccharification and fermentation were simultaneously performed at 30 o C with yeast in the supporting of liquefying and saccharifying enzymes. Moreover, the WDG was differently harvested and treated from the factory to others. The WDG was separated by a plate-frame filter before the alcohol distillation to avoid blocking the distilling tower as at Factory 1 and Factory 2. Meanwhile, in other factories, the WDGs were harvested from the whole stillage after the alcohol distillation by a high-speed decanter (Factory 3, Factory 4) or by frame-plate filter (Factory 5). Moreover, in Factory 3, the thin stillage after the decantation was partly reused to reduce the pH of the rice mash. Interestingly, in Factory 4, spent from the cassava starch harvesting process, containing fiber up to 50% dry matter, was combined with WDG from the bioethanol process. The composition of raw materials such as white rice, cassava chips and corn has been extensively elucidated. Based on dry matter, rice (milled rice or white rice) is composed of less crude fiber (0.1–0.8%) than corn (3%) or cassava (3.7- 4.0%) [ 18 – 21 ]. On the other hand, white rice contains 7–8% of crude protein, which is less than that in corn (9.42%) and higher than in cassava (1–3%) [ 18 – 21 ]. Three types of raw materials were composed of a comparable level of carbohydrates with mainly starch, i.e., 80% for rice and cassava and 72% for corn. During the bioethanol processing, the un-converted components such as crude fiber in the raw material, which are mostly left in DDG, could lead the difference in DDG composition. This explains the much higher level of protein in the rice DDG than that in the corn DDG and the cassava DDGs (Table 2 ). Unlike wheat/corn DDGS, even though the soluble in thin stillage was not taken into account (Fig. 1 ), the rice DDGs in this study contained more protein than the corn DDGS which ranged from 26–32% [ 2 , 17 , 22 – 24 ]. Also, in this work the protein content in the corn DDG (Factory 5-CO) was slightly higher than the range (35.6%) (Table 2 ). Yeast biomass has been known as a rich source of protein which was reported in a range of 38.8–70.7% dry matter [ 25 ]. The amount of protein in DDGS is affected by yeast as shown in a review by Liu et al [ 2 ]. In an uncertain estimation, the authors indicated that yeast contributes approximately 5.3% of the protein in DDGS [ 2 ]. This number is much lower than the amount of protein observed in rice DDG in this study (50–80% of dry matter). This suggests most of the protein in rice DDG was from rice material. This statement is also supported by the low protein value in cassava DDGs (13.38–16.40%) (Table 2 ). If protein in DDG were mainly contributed by yeast biomass, the protein content in cassava-DGG would be much higher than the observed values. The higher fiber in cassava resulted in the higher of this parameter in cassava DDGs (up to 36.5% in Factory 5-CS) in comparison to the rice DDGs (Table 2 ). The high content of fiber in the cassava DDG Factory 4-CS was explained by the addition of the cassava spent by-product of starch processing which contains up to 50% of fiber. The fiber content in the corn DDG (Factory 5-CO) in this study was in agreement with the reported range for DDGS (7.22–10.2%) [ 1 , 17 , 24 ]. The content of residual starch in DDG samples indicated the effectiveness of heat treatment, amylolytic conversion by amylases and fermentation by yeast [ 17 ]. Starch content in corn DDGS ranges from 3–6% [ 2 , 17 ]. The starch content in DDGs in this study might be overestimated due to the analysis method using HCl hydrolysis. The reducing sugar from non-starch carbohydrates can be generated after acid hydrolysis. Despite the lower ash content in cassava [ 18 , 20 ], the ash content of cassava DDG was higher than that in rice DDG (1.44–2.61%) and corn DDG (3%) (Table 2 ). There was no correlation between fat content in raw material and DDG samples. For example, the fat content in rice (1–2%) and cassava (0.5-1.0%) is lower than in maize (4.72%) [ 19 , 20 ], and the fat in the rice-DDG varied from 0.69% − 10.41% (Table 2 ). We also found in this work that the fat content in the corn DDG Factory 5-CO was 4.56%, which is half to one-third of the reported range from 10.2–14.5% of DDGS [ 2 , 17 , 23 , 24 ]. The difference in compositions of the rice DDG samples from Factory 2-R, Factory 3-R, Factory 4-R and the SLSF-R suggests that the ethanol processing technology leads to a significant effect. Especially, the Factory 2-R and the SLSF-R used the same rice material but different process technology showing varied data in protein, starch, fiber and other components. Interestingly, the SLSF-R differs remarkably from that of Factory 2-R on protein (51.49%), fiber (18.42%) and fat (7.47%). It is likely because of the uncooked and simultaneous saccharification and fermentation in the SLSF process (Fig. 1 ). 4.2 The potential of rice in food application and cassava DDG in animal feeding DDGS was considered not only for animal feeding but also for food application recently. This study showed that the rice DDG which contained protein up to 50–80% of dry matter was highly potential for food application, especially as the source of plant protein. In comparison to soybean seed which is well-known as a main source of plant protein for food application currently, the protein in rice DDG was much higher than the protein in soybean seed (ca 40% of dry matter [ 26 ]). Moreover, the profile of amino acids in the rice DDG is balancing of essential and non-essential amino acids (Table 3 ). Similarly in rice as reported previously [ 27 ], in the rice DDG the essential amino such as isoleucine, leucine, phenylalanine, valine, threonine are also present at higher levels than others. This also means the ethanol process did not change the amino acid profile of raw material to DDG as mentioned previously [ 2 ]. Lysine, an important amino acid, is considered the first limiting in cereals such as maize, cassava and even in some varieties of rice [ 28 ]. In some varieties of Vietnamese rice, lysine was in the range of 3–5% of total protein [ 27 ], which is in agreement with the lysine content in rice DDGs found in this study (approximately 2.5–4% of rice DDG) (Table 3 ). Lysine presents in rice DDG at a level of 2.5-4 times higher than in cassava or corn DDG (Table 3 ). Among rice DDGs, SLSF-R was not treated with heating, therefore protein (~ 51% of dry matter) in the SLSF-R was expectedly unmodified as in other rice DDGs due to heat treatment. Protein from SLSF-R was preliminarily harvested with alkaline extraction and precipitation by pH 4.5 adjustment. The main protein fraction in rice, glutenin (amounted to 75–90% of total rice protein) [ 29 ], was reported as well-dissolved in alkaline condition [ 30 ], therefore NaOH 0.5M was used for extraction. The protein content was increased to 70–72% in resulted precipitate with a recovery yield of 54–55% (Table 4 ). The enriched protein precipitate from SLSF-R was applied to replace wheat flour in the bread recipe. With such high protein, the replacement of enriched-protein precipitate from SLSF-R to wheat flour can improve the protein content in bread, however, it led the reduction of gluten which is needed for the textural structure of bread, therefore the rheological properties of bread were negatively affected, especially at the high ratio of replacement [ 3 ]. Among these, the ratio of 5% replacement was acceptable in flavor and taste. Even though, with a such high protein content, it is highly potentially applied in many non-structural food products. Protein from rice DDG, furthermore, could be used for enzymatic hydrolysis to improve the digestibility [ 13 ] or to obtain the bioactive peptides [ 31 ]. Cassava DDG contained a low level of protein but a high level of fiber. Fiber, even though does not generate energy, has to be included in the diet to maintain normal physiological functions in the digestive tract [ 32 ]. Using molds for fermentation to enrich protein in cassava DDG has been reported by Mai-Dinh Vuong et al. with protein content increased from 11.96 to 15.27% using Trichoderma harzianum strain [ 11 ], similar results were observed in this study with the protein content increased from the initial value of 16.42% up to 18.08% − 20.7% of dry matter, depending on strains (Table 6 ). Among used fungal strains, Rhizomucor miehei FCH 116.3 and Thermothelomyces thermophilus FCH 112.2 showed better protein improvement than others. Moreover, the evaluation of carbohydrate-active enzymes showed that the strains had the ability to produce the three enzymes required in animal feed on cassava-based WDG substrate (Fig. 3 ). The on-site production of enzymes through fermentation for animal feed production and high protein biomass generation is a promising strategy. 5. Conclusions In this study, the first systematic analysis of proximates of rice/cassava-based DDG was performed and showed the variation of proximates in DDG samples which depended on bioethanol technology, WDG treatments as well as raw materials. Among them, the rice DDGs with the high protein content and balancing amino acid profile were shown as the highly potential plant protein source for food application. Meanwhile, cassava DDG also exhibited valuable for animal feeding not only due to the high level of fiber but also the substrate source for accumulation of the animal health beneficial enzymes by solid-state fermentation with fungi. Abbreviations CMC Carboxymethylcellulose DDG Distillers dried grain DDGS Distillers dried grain with soluble DNS 3,5-dinitrosalicylic acid PDA Potato Dextrose Agar SLSF Simultaneous liquefaction saccharification and fermentation process WDG Wet distillers grain WDGS Wet distiller’s grains with soluble Declarations Conflict of interest The authors declare that they have no conflict of interest. Funding This work was supported by project NVQG2020/ĐT.01, Ministry of Science and Technology, Vietnam Author contributions Tien-Thanh Nguyen performed the experiments and wrote the manuscripts. Bach Cao-Xuan performed experiments and drafted partly the manuscripts. Pham Kim Dang, Thuy Nguyen-Thanh, Tien Cuong Nguyen revised partly the manuscripts. Nguyen Gia Long, Mai Dinh Vuong performed the experiments. Chu-Ky Son, Vu Nguyen Thanh supervised, finalized, and submitted. Data availability The datasets are available from the corresponding author on reasonable request. References Monceaux, D. and D. Kuehner: Dryhouse technologies and DDGS production. Lallemand ethanol technology and Nottingham University Press, (2009). Liu, K.: Chemical composition of distillers grains, a review . J. Agri. Food Chem., 59(5), 1508-1526 (2011). Li, X., C. Wang, and P.G. Krishnan: Effects of corn distillers dried grains on dough properties and quality of Chinese steamed bread . Food Sci. Nutr., 8(8), 3999-4008 (2020). Pourafshar, S., K.A. Rosentrater, and P.G. Krishnan: Changes in chemical and physical properties of Latin American wheat flour based tortillas substituted with different levels of distillers dried grains with solubles (DDGS) . J. Food Sci. Technol., 52(8), 5243-5249 (2015). Pourafshar, S., K.A. Rosentrater, and P.G. Krishnan: Production of Barbari bread (traditional Iranian bread) using different levels of distillers dried grains with solubles (DDGS) and sodium stearoyl lactate (SSL) . Foods, 7(3), 31 (2018). Rosentrater, K.: Incorporating distillers grains in food products . Cereal Foods World - CFW, 51, 52-60 (2006). Singha, P., K. Muthukumarappan, and P. Krishnan: Influence of processing conditions on apparent viscosity and system parameters during extrusion of distiller's dried grains-based snacks . Food Sci. Nutr., 6(1), 101-110 (2018). Ministry-of-Trade-and-Industry-of-Vietnam: Targets of the scheme to develop of beverage in Vietnam to 2025 and the vision to 2035 (Decision No 3690/QĐ-BCT). 2016: 12/06/2016. Xuan, D.N.N., M.L. Huu, and U. Peter: Tropical fibre sources for pigs—digestibility, digesta retention and estimation of fibre digestibility in vitro . Anim. Feed Sci. Technol., 102(1–4), 109-124 (2002). Taranu, I., et al.: Rice and cassava distillers dried grains in Vietnam: Nutritional values and effects of their dietary inclusion on blood chemical parameters and immune Responses of Growing Pigs . Waste and Biomass Valorization, 10(11), 3373-3382 (2019). Dinh Vuong, M., et al.: Protein enrichment of cassava-based dried distiller’s grain by solid state fermentation using Trichoderma harzianum and Yarrowia lipolytica for feed ingredients . Waste and Biomass Valorization, 12, 3875-3888 (2021). Miller, G.L.: Use of dinitrosalicylic acid reagent for determination of reducing sugar . Analytical Chemistry, 31(3), 426-428 (1959). Do, T.-T.-H., et al.: Extraction and hydrolysis of protein from industrial rice-based dried distiller’s grain toward food application (origin text in Vietnamese) . J. Vietnam Agri. Sci. Technol., 01(122), 106 - 112 (2021). Thanh, V.N., et al.: Surveying of acid-tolerant thermophilic lignocellulolytic fungi in Vietnam reveals surprisingly high genetic diversity . Scientific Reports, 9(1), 3674 (2019). Alagón, G., et al.: Nutritive value of distillers dried grains with solubles from barley, corn and wheat for growing rabbits . Anim. Feed Sci. Technol., 222, 217-226 (2016). Belyea, R.L., et al.: Sources of variation in composition of DDGS . Anim. Feed Sci. Technol., 159(3), 122-130 (2010). Belyea, R.L., K.D. Rausch, and M.E. Tumbleson: Composition of corn and distillers dried grains with solubles from dry grind ethanol processing . Bioresour. Technol., 94(3), 293-298 (2004). Montagnac, J.A., C.R. Davis, and S.A. Tanumihardjo: Nutritional value of cassava for use as a staple food and recent advances for improvement . Compr. Rev. Food Sci. F., 8(3), 181-194 (2009). Nuss, E.T. and S.A. Tanumihardjo: Maize: A paramount staple crop in the context of global nutrition . Compr. Rev. Food Sci. F., 9(4), 417-436 (2010). Morgan, N.K. and M. Choct: Cassava: Nutrient composition and nutritive value in poultry diets . Anim. Nutri., 2(4), 253-261 (2016). Zhou, Z., et al.: Composition and functional properties of rice . Inter. J. Food Sci. Technol., 37(8), 849-868 (2002). Cromwell, G.L., K.L. Herkelman, and T.S. Stahly: Physical, chemical, and nutritional characteristics of distillers dried grains with solubles for chicks and pigs . J. Anim. Sci., 71(3), 679-686 (1993). Kim, Y., et al.: Composition of corn dry-grind ethanol by-products: DDGS, wet cake, and thin stillage . Bioresour. Technol., 99(12), 5165-5176 (2008). Spiehs, M.J., M.H. Whitney, and G.C. Shurson: Nutrient database for distiller's dried grains with solubles produced from new ethanol plants in Minnesota and South Dakota . J. Anim. Sci. , 80(10), 2639-2645 (2002). Martini, A.E.V., M.W. Miller, and A. Martini: Amino acid composition of whole cells of different yeasts . J. Agri. Food Chem., 27(5), 982-984 (1979). Molinari, M.D., et al.: Exploring the proteomic profile of soybean bran: Unlocking the potential for improving protein quality and quantity . Plants, 12(14)(2023). Khoi, B.H., et al.: The protein and the amino acid composition of some rice and maize varieties grown in North Vietnam . J. Sci. Food Agri., 39(2), 137-143 (1987). Torbatinejad, N.M., S.M. Rutherfurd, and P.J. Moughan: Total and reactive lysine contents in selected cereal-based food products . J. Agri. Food Chem., 53(11), 4454-4458 (2005). Kawakatsu, T. and F. Takaiwa: 4 - Rice proteins and essential amino acids. Rice (Fourth Edition), ed. J. Bao. AACC International Press, (2019). Cookman, D.J. and C.E. Glatz: Extraction of protein from distiller’s grain . Bioresource Technology, 100(6), 2012-2017 (2009). Selamassakul, O., et al.: Bioactive peptides from brown rice protein hydrolyzed by bromelain: Relationship between biofunctional activities and flavor characteristics . J Food Sci, 85(3), 707-717 (2020). Lindberg, J.E.: Fiber effects in nutrition and gut health in pigs . J. Anim. Sci. Biotechnol., 5(1), 15 (2014). 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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-3171967","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":275918302,"identity":"fce3cd24-e21a-44ab-86d7-3bcb31979d9c","order_by":0,"name":"Tien-Thanh Nguyen","email":"","orcid":"","institution":"Hanoi University of Technology: Hanoi University of Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Tien-Thanh","middleName":"","lastName":"Nguyen","suffix":""},{"id":275918303,"identity":"1956857f-be59-497a-bb67-a63ae5c0d47e","order_by":1,"name":"Gia Long Nguyen","email":"","orcid":"","institution":"Hanoi University of Technology: Hanoi University of Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Gia","middleName":"Long","lastName":"Nguyen","suffix":""},{"id":275918304,"identity":"86f1e4f9-d469-44b9-aa92-09ef88698f69","order_by":2,"name":"Bach Cao-Xuan","email":"","orcid":"","institution":"Food Industries Research Institute","correspondingAuthor":false,"prefix":"","firstName":"Bach","middleName":"","lastName":"Cao-Xuan","suffix":""},{"id":275918305,"identity":"06e7174a-3112-4dee-bdcc-6bdd38db5087","order_by":3,"name":"Thuy Nguyen-Thanh","email":"","orcid":"","institution":"Food Industries Research Institute","correspondingAuthor":false,"prefix":"","firstName":"Thuy","middleName":"","lastName":"Nguyen-Thanh","suffix":""},{"id":275918306,"identity":"12dd4793-b755-4f61-9a09-11c02b7d38bc","order_by":4,"name":"Vuong Mai Dinh","email":"","orcid":"","institution":"Hanoi University of Technology: Hanoi University of Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Vuong","middleName":"Mai","lastName":"Dinh","suffix":""},{"id":275918307,"identity":"7b82897a-05b9-405e-a9ad-2d7f8f1524c2","order_by":5,"name":"Tien Cuong Nguyen","email":"","orcid":"","institution":"Hanoi University of Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Tien","middleName":"Cuong","lastName":"Nguyen","suffix":""},{"id":275918308,"identity":"1aa556cf-6beb-4ec5-b6dc-0a1f4b8b0d09","order_by":6,"name":"Kim Dang Pham","email":"","orcid":"","institution":"Vietnam National University of Agriculture","correspondingAuthor":false,"prefix":"","firstName":"Kim","middleName":"Dang","lastName":"Pham","suffix":""},{"id":275918309,"identity":"e8183b05-b41d-4ed7-92da-d799dba167d5","order_by":7,"name":"Nguyen Thanh Vu","email":"","orcid":"","institution":"Food Industries Research Institute","correspondingAuthor":false,"prefix":"","firstName":"Nguyen","middleName":"Thanh","lastName":"Vu","suffix":""},{"id":275918310,"identity":"8574e1ad-b65d-4f2c-a4cf-8d9dd555af98","order_by":8,"name":"Son Chu-Ky","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA5UlEQVRIiWNgGAWjYLACxgYGAwb2HijvANFaeM4AWQkkaZHIIVKLfHvvAYafOw4bG9x8e0zy5w8GOb4bCWzSBXi0GJw5l8DYe+awmcHtvDRpngQGY0mQlhn4tEjkGDAzth22MbidYyYNdFjihhsJzMY8+Bw2A6bl5hkzyR8JDPUEtTDcgGgxM7jBYyYBdFiCwY0Exsf4tBicOWNwsLct3VjyTI6xNU+ahOHMMw8b8WqRb+8xfPCzzdqw7/gZw5s/bGzk+Y4nHziM12EM0IhQAJMMEgzgaCIKyBOpbhSMglEwCkYgAABvbEwzv3CvkwAAAABJRU5ErkJggg==","orcid":"https://orcid.org/0000-0001-6574-1326","institution":"Hanoi University of Science and Technology","correspondingAuthor":true,"prefix":"","firstName":"Son","middleName":"","lastName":"Chu-Ky","suffix":""}],"badges":[],"createdAt":"2023-07-15 02:50:26","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-3171967/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-3171967/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":52022276,"identity":"a60ca55c-af82-406d-899b-2cbf0c3762ad","added_by":"auto","created_at":"2024-03-05 15:04:52","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":55408,"visible":true,"origin":"","legend":"\u003cp\u003eEthanol production outlines applied in 5 factories. a. Factory 1, Factory 2; b. Factory 3, Factory 4; c. Factory 5; d. SLSF (from factory 2)\u003c/p\u003e","description":"","filename":"1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3171967/v1/38cf38a37268fc020689c080.jpg"},{"id":52022279,"identity":"06689f1d-c47e-43e0-b178-6b77f8cd9a8e","added_by":"auto","created_at":"2024-03-05 15:04:52","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":42930,"visible":true,"origin":"","legend":"\u003cp\u003eBread samples with different replacement ratio of the enriched protein precipitate for wheat flour\u003c/p\u003e","description":"","filename":"2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3171967/v1/01eeb640edd689247674f5fd.jpg"},{"id":52022278,"identity":"570fcc91-33cb-4a81-b687-1612578a8045","added_by":"auto","created_at":"2024-03-05 15:04:52","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":63721,"visible":true,"origin":"","legend":"\u003cp\u003eEnzyme activities in the solid-state cultivation of the 4 fungal strains on the cassava DDG Factory 5-CS\u003c/p\u003e","description":"","filename":"3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3171967/v1/5b039d2e397b755161bddfac.jpg"},{"id":53812647,"identity":"2da24648-9c35-4249-a241-5b52fd316202","added_by":"auto","created_at":"2024-03-31 15:16:47","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":589190,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3171967/v1/ee04961f-da4a-499b-b5ae-70a1dc8244a2.pdf"},{"id":52022718,"identity":"c00353a4-a997-41d6-ad49-e50044d63771","added_by":"auto","created_at":"2024-03-05 15:12:52","extension":"jpg","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":40697,"visible":true,"origin":"","legend":"","description":"","filename":"Graphicabstract.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3171967/v1/71a836d2a86f1a2f164f4509.jpg"}],"financialInterests":"","formattedTitle":"\u003cp\u003eRice-Based Distillers’ Dried Grains from Bio-Ethanol Production, a Potential Source of Protein for Food Application \u003c/p\u003e","fulltext":[{"header":"Highlights ","content":"\u003cul\u003e\n \u003cli\u003eThe first systematic analysis of composition of DDG from rice and cassava- based bioethanol\u0026nbsp;factory show the variation of proximates depending on technologies and raw materials\u003c/li\u003e\n \u003cli\u003eRice DDG is\u0026nbsp;high\u0026nbsp;potential of\u0026nbsp;non-animal\u0026nbsp;protein for food application.\u003c/li\u003e\n \u003cli\u003eCassava DDG can be valorized by solid state fermentation with thermostable fungi to improve protein content and accumulate beneficial enzymes for animal digestion system\u0026nbsp;\u003c/li\u003e\n\u003c/ul\u003e"},{"header":"1. Introduction","content":"\u003cp\u003eDistiller\u0026rsquo;s dried grain with soluble (DDGS) has been known as a co-product of the bioethanol industry from corn or wheat [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. The composition of DDGS has been received a great interest in animal science, ethanol producers, and especially in the animal feeding industry as most of this co-product has been sold as feed ingredients for livestock [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. DDGS was also considered as a potential source of insoluble dietary fiber which could be beneficial in the prevention of colon cancer, coronary heart disease, diabetes, and obesity. Therefore, DDGS was recently valorized as an ingredient for food application [\u003cspan additionalcitationids=\"CR4 CR5 CR6\" citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eBioethanol including for biofuel and beverage production is highly demanded in Vietnam [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. The expected production of bioethanol in Vietnam is about 170\u0026nbsp;million liters for the food industry by 2035 [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. There are several bioethanol factories using different production technologies with rice or cassava as starchy materials. Regardless of raw materials, it is estimated that to gain 1 liter of ethanol, 2.3 kg of material are used and about 0.15 kg of dry matter (6.5% of raw material) of distiller grain are produced. This means that by 2035, approximately 0.65\u0026nbsp;million tons of dried distiller grain will be generated in Vietnam.\u003c/p\u003e \u003cp\u003eThe wet distiller grain (WDG) and dried distiller grain (DDG) from ethanol factories has been used since a long time ago for the purpose of feeding in Vietnam. These by-products usually are directly added to the diet of animals without or with simple treatment [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. Few studies have started to focus on cassava and rice DDG, mainly targeting animal feeding. For example, authors used rice and DDG in pig feed and showed beneficial effects on animal performance [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. Cassava DDG has been improved the protein content by solid fermentation with yeasts or molds [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. However, the potential of these DDGs, especially rice-DDG could be extended because rice is used as the staple food in Vietnam and many Asian countries.\u003c/p\u003e \u003cp\u003eThis work aimed to elucidate systematically the nutrient composition of DDG samples from different bioethanol factories in Vietnam which use different technologies on either cassava or rice as raw materials. Furthermore, the protein content of DDG, especially rice based DDG was preliminarily improved and applied in food products. The results showed the high potential of DDG from rice-based bioethanol in the food industry.\u003c/p\u003e"},{"header":"2. Materials and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1 Samples collection\u003c/h2\u003e \u003cp\u003eSeven wet distillers spent (WDS) samples were collected from 5 ethanol factories located over the country (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The WDS samples were transported immediately after production directly to Hanoi University of Science and Technology (Vietnam) and were dried following a protocol as follows: 90\u003csup\u003eo\u003c/sup\u003eC for 30 min, 80\u003csup\u003eo\u003c/sup\u003eC for 2.5-3 h and finally 70\u003csup\u003eo\u003c/sup\u003eC for 1 h in a circulating dryer. The dried distiller grain (DDG) samples then were packed in plastic bags and stored at -10\u003csup\u003eo\u003c/sup\u003eC for further analysis. Depending on the raw material for bioethanol production, the samples were noted with R (rice), CS (cassava) or C (corn). Another sample (named SLSF) was obtained from a simultaneous liquefaction, saccharification and fermentation process (SLSF) conducted at factory 2 in the Northern with rice (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ed).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eOverview of ethanol production and wet distillers\u0026rsquo; grain (WDG) in 5 factories in Vietnam\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"8\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eSamples\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSLSF\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eFactory 1\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eFactory 2\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eFactory 3\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eFactory 4\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eFactory 5-CS\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eFactory 5-CO\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eCapacity (L/year)\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e150,000\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1,500,000\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1,200,000\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e10,000,000\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e100,000,000\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e76,000,000\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e76,000,000\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eEnd user\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBeverage ethanol\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eBeverage ethanol\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eBeverage ethanol\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eBeverage ethanol\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eFuel ethanol\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eFuel ethanol\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eBeverage ethanol\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eRaw materials\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eRice\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eRice\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eRice\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eRice\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eCassava\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eCassava\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eCorn\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eProcessing\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSimultaneous liquefaction, saccharification and fermentation at pilot scale (500 L/batch)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eSeparate liquefaction, saccharification and fermentation\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eSeparate liquefaction, saccharification and fermentation\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eSeparated Liquefaction Simultaneous Saccharification and fermentation\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eSeparated Liquefaction Simultaneous Saccharification and fermentation\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eLiquefaction, saccharification and continuous fermentation\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eLiquefaction, saccharification and continuous fermentation\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eSeparation\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eWet distillers\u0026rsquo; grains (WDG) were filtered before distillation by a frame and plate filtration\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eWDG were filtered before distillation by a frame and plate filtration\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eWDG were filtered before distillation by a frame and plate filtration\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eWDG were separated after distillation by decanter\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eWDG were separated after distillation by decanter\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eWDG were filtered after distillation by a frame and plate filtration\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eWDG were filtered after distillation by a frame and plate filtration\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eDrying\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eFlash drying\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eFlash drying\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eFlash drying\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2. Proximate composition analysis\u003c/h2\u003e \u003cp\u003eProximate composition in DDG samples was analyzed by using the standard methods including lipid (ISO 6492:1999), ash (ISO 05984:2002), crude fiber (with Ankom filter bag technique), total crude protein (ISO 05983-1:2005), phosphorus (ISO 6491:1998), calcium (ISO 06490:1985). Starch was determined via reducing sugar generated by hydrolyzing samples in HCl 2% for 2 hours in a boiling water bath. The reducing sugar content was determined by using dinitrosalicylic acid [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3 Amino acid profile determination\u003c/h2\u003e \u003cp\u003eThe amino acid profile in DDG samples was determined as previously described. Shortly, samples (ca. 40 mg) were hydrolyzed with vapor of HCl 6M, 0.5% phenol for 24h at 120\u003csup\u003eo\u003c/sup\u003eC. The amino acids in the neutralized hydrolysates were derivatized with OPA, separated on the column C18 ElipseZorbax 5 \u0026micro;m, 4.6 x 150 mm (Agilent, US) and detected by DAD detector of Agilent 1200 series HPLC (USA). Two mobile phases, sodium phosphate 40 mM, pH 7.8 and methanol: acetonitrile: deionized water\u0026thinsp;=\u0026thinsp;45:45:10, were used in the 31-minute gradient elution program as previously described [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e2.4. Preliminary valorization of rice DDG and cassava DDG\u003c/h2\u003e \u003cp\u003e \u003cem\u003eExtraction and application of protein in rice SLSF DDG\u003c/em\u003e \u003c/p\u003e \u003cp\u003eProtein in rice-SLSF DDG was extracted by using the alkaline solution as described previously [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. The rice DDG from SLSF was mixed at the ratio of 1:30 weight/volume with NaOH 0.5M and NaHSO\u003csub\u003e3\u003c/sub\u003e 50 mM. The mixtures were stirred at 600 rpm and maintained at different temperatures of 25\u003csup\u003eo\u003c/sup\u003eC, 50\u003csup\u003eo\u003c/sup\u003eC and 70\u003csup\u003eo\u003c/sup\u003eC with a water bath (Memmert WNB 22, Germany) for 2 hours. Mixtures then were centrifuged at 6000 \u0026times;g/15 min to separate supernatants and residue solids. The residue solids were washed with water, dried, and determined the dry weight and the protein content. The supernatants were adjusted pH to 4.0 with HCl to precipitate the extracted protein. Precipitates were collected by a similar centrifuging step. Similarly, the precipitates were washed with water, dried, and used for measurement of protein.\u003c/p\u003e \u003cp\u003eThe precipitate which was enriched in protein content then was preliminarily applied to bread recipe by replacement of wheat flour. The ratio of replacement was from 5\u0026ndash;20%. Bread quality parameters including rheological/sensorial parameters such as hardness, cohesiveness, resilience, chewiness, springiness, gumminess were measured by using a TA.XT \u0026ndash; plus texture analyzer (Stable Micro Systems, UK) with 5 kg loadcell sensor, 25mm diameter probe (P/25). The texture profile of the bread was measured 1 hour after baking. The breads were sliced at the largest circumference with slice thickness of 20 mm. The speed was of 5mm/s and the compressive deformation was 50% (10mm).\u003c/p\u003e \u003cp\u003e \u003cem\u003eSolid-state cultivation of cassava DDG by thermophilic fungi\u003c/em\u003e \u003c/p\u003e \u003cp\u003eCassava DDG (Factory 5-CS) was used for solid-state fermentation with four thermophilic fungal strains including \u003cem\u003eRhizomucor miehei\u003c/em\u003e FCH 116.3; \u003cem\u003eRhizopus microsporus\u003c/em\u003e LPH 084; \u003cem\u003eRhizopus microsporus\u003c/em\u003e LPH 156; \u003cem\u003eThermothelomyces thermophilus\u003c/em\u003e FCH 112.2 [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eFungal spores on PDA plates were collected after 5 days of cultivation at 50\u0026deg;C and suspended in 4 mL of sterile saline 0.9% w/v and 0.05% Tween 80 and used as inoculums. Two milliliters of each inoculum were added into cassava WDG (~\u0026thinsp;5 g dry matter) in a 500 mL Erlenmeyer flask. After 7 days of incubation at 50\u0026deg;C, the protein content and enzyme activities (xylanase, CMCase, amylase) in solid phases were measured. For protein measurement, the dried solid was used as described above. For enzyme extraction, 50 mM sodium citrate buffer pH 5.0 was added to the solid medium in flasks and mixed for 1 h at 30\u0026deg;C at 160 rpm. The supernatant was obtained by centrifugation at 6000 \u0026times;g/10 min at 10\u0026deg;C and stored at \u0026minus;\u0026thinsp;20\u0026deg;C until analysis. For the determination of xylanase, CMCase and amylase activity, the extracts were filtrated through 0.45 \u0026micro;m filter (Minisart\u0026reg; NY25 Syringe Filter) and used for enzyme reaction with substrates including 1% xylan (xylan from beechwood, Apollo Scientific\u0026reg;), CMC (Sigma-Aldrich) and Starch (Samchun chemicals\u0026reg;) in 50 mM sodium citrate pH 5 buffer, respectively. The enzyme reactions were performed at 50\u003csup\u003eo\u003c/sup\u003eC for 20 min including 0.1 ml of extract and 0.2 ml of substrate solution. The enzymatically generated reducing sugar concentrations were measured by using dinitrosalicylic acid (DNS) as mentioned above.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003e2.6 Data analysis\u003c/h2\u003e \u003cp\u003eDescriptive statistical parameters such as mean values and standard errors were calculated using Microsoft Excel from at least duplicate measurements.\u003c/p\u003e \u003c/div\u003e"},{"header":"3. Results","content":"\u003cp\u003eSeven WDS samples were collected from 5 factories that are different in raw materials, capacity, ethanol production process and WDG harvesting and treatment (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Among, 4 samples, named SLSF-R, Factory 1-R, Factory 2-R and Factory 3-R, were from rice material (rice-based); 2 samples Factory 4-CS and Factory 5-CS were from cassava material (cassava-based); and the rest (Factory 5-CO) was based on corn material. Seven DDG samples were obtained after drying, respectively. These DDG samples were analyzed for the proximate components and amino acid profiles.\u003c/p\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003e3.1. Proximate composition of rice-based and cassava DDG samples\u003c/h2\u003e \u003cp\u003eDDG samples obtained from 5 factories were different in proximate composition as shown in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eComposition of Dried Distillers Grain (DDG) samples\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"10\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003eCompositions\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eSLSF-R\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eFactory 1-R\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eFactory 2-R\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eFactory 3-R\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eFactory 4-CS\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eFactory 5-CS\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003eFactory 5-CO\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c10\"\u003e \u003cp\u003eRef DDGS\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003eRaw materials\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eRice\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eRice\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eRice\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eRice\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eCassava\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eCassava\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eCorn\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eCorn\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003eCrude Protein (% DM)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e51.49\u0026thinsp;\u0026plusmn;\u0026thinsp;0.84\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e72.44\u0026thinsp;\u0026plusmn;\u0026thinsp;1.36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e74.96\u0026thinsp;\u0026plusmn;\u0026thinsp;2.40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e79.60\u0026thinsp;\u0026plusmn;\u0026thinsp;1.13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e13.38\u0026thinsp;\u0026plusmn;\u0026thinsp;0.64\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e16.40\u0026thinsp;\u0026plusmn;\u0026thinsp;0.72\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e35.60\u0026thinsp;\u0026plusmn;\u0026thinsp;0.82\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e31.40\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"3\" rowspan=\"4\"\u003e \u003cp\u003eNon-protein (% DM)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStarch\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e15.01\u0026thinsp;\u0026plusmn;\u0026thinsp;0.17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e14.29\u0026thinsp;\u0026plusmn;\u0026thinsp;0.57\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e11.77\u0026thinsp;\u0026plusmn;\u0026thinsp;0.51\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e8.55\u0026thinsp;\u0026plusmn;\u0026thinsp;0.38\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e19.43\u0026thinsp;\u0026plusmn;\u0026thinsp;0.86\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e10.90\u0026thinsp;\u0026plusmn;\u0026thinsp;0.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e26.63\u0026thinsp;\u0026plusmn;\u0026thinsp;1.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e5.30\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCrude Fiber\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e18.42\u0026thinsp;\u0026plusmn;\u0026thinsp;0.82\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e9.09\u0026thinsp;\u0026plusmn;\u0026thinsp;0.27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e11.58\u0026thinsp;\u0026plusmn;\u0026thinsp;0.42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e8.98\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e46.72\u0026thinsp;\u0026plusmn;\u0026thinsp;1.70\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e36.50\u0026thinsp;\u0026plusmn;\u0026thinsp;1.70\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e7.59\u0026thinsp;\u0026plusmn;\u0026thinsp;0.21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e10.20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFats\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e7.47\u0026thinsp;\u0026plusmn;\u0026thinsp;0.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e10.41\u0026thinsp;\u0026plusmn;\u0026thinsp;0.42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.69\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2.30\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e2.94\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e4.56\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e12.00\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eAsh\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.74\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.27\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.44\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2.61\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e9.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e12.08\u0026thinsp;\u0026plusmn;\u0026thinsp;0.42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e3.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e4.60\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003eTotal non-protein (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e42.64\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e36.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e26.90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e20.83\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e77.50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e62.42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e41.78\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e32.10\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003eCalcium (% DM)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.41\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003ePhosphorus (% DM)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"3\" nameend=\"c3\" namest=\"c1\"\u003e \u003cp\u003e\u003cem\u003eR: rice; CS: cassava; CO: corn\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe protein in rice DDG amounted to 50\u0026ndash;80% of dry matter, while in cassava DDG, this value was only 13.38\u0026ndash;16.40% of dry matter. The protein composition in the corn DDG Factory 5-CO was 35.6% of dry matter.\u003c/p\u003e \u003cp\u003eThe starch content was measured in a range of 8.55\u0026ndash;15.01% of dry matter for the rice DDG, and 10.90-19.43% for the cassava DDG. Meanwhile, the starch contributed up to 26.63% of dry matter in the corn DDG sample.\u003c/p\u003e \u003cp\u003eThe fiber content was found inconsistent among these DDGs. The cassava DDG (Factory 4-CS and Factory 5-CS) contained significantly high fiber, which was 46.72% for the former and 36.5% for the latter. The fiber content of the rice DDGs varied in a range of 9.00\u0026ndash;11.58%, except for the SLSF-R which was 18.42%. The corn DDG Factory 5-CO contained a slightly lower fiber compared to the rice DDG, amounting to 7.59% of dry matter.\u003c/p\u003e \u003cp\u003eFat content was found to rather varied from sample to sample. Fat was at a higher level in the rice DDG such as SLSF-R (7.47%), Factory 1-R (10.41%). Surprisingly, the DDG Factory 3-R contained rather low fat (0.69%). The fat in cassava DDGs varied in a narrow range of 2.30\u0026ndash;2.94% of dry matter. The fat content in corn DDG Factory 5-CO was 4.56% of dry matter\u003c/p\u003e \u003cp\u003eThe ash of rice DDG samples amounted to 1.44\u0026ndash;2.61% of dry matter. This component was slightly higher in the corn DDG (3%). In contrast, ash contributed a high level in the cassava DDG samples ranging from 9.05 to 12.08%.\u003c/p\u003e \u003cp\u003eCalcium and phosphorus were also figured out for the rice DDGs and one cassava DDG sample. Phosphorus was at a low level in all the test samples (between 0.01\u0026ndash;0.03% of dry matter). Calcium was very low in the rice DDG Factory 1-R (0.02%), but much higher in the cassava DDG Factory 4-CS.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003e3.2. Amino acid profiles\u003c/h2\u003e \u003cp\u003eTable\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e shows the consistent results on amino acid profiles. The amino acid profile was generally similar among the DDG samples which derived from the same type of raw material. In agreement with the protein composition, the amino acid was lower in cassava and corn DDG than in rice DDG samples. Most of the essential amino acids with significant amounts were determined in the DDG, especially the rice DDG samples. Leucine, phenylalanine, arginine, lysine contributed more than 3% of dry matter in the rice DDGs. These amino acids amounted much lower in cassava and corn DDGs, except leucine in the corn DDG Factory 5-CO sample which was comparable to that of rice DDG (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eProfile of amino acids of DDG samples\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"8\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAmino acids (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSLSF-R\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eFactory 1-R\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eFactory 2-R\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eFactory 3-R\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eFactory 4-CS\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eFactory 5- CS\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eFactory 5-CO\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colspan=\"8\" nameend=\"c8\" namest=\"c1\"\u003e \u003cp\u003e\u003cem\u003eEssentials amino acids\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHIS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.65\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.35\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.64\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.31\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.27\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.93\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eARG\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.60\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.41\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4.44\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e3.44\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.22\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.21\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.91\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTHR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.87\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.17\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.23\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.28\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.50\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.70\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.99\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eVAL\u0026thinsp;+\u0026thinsp;MET\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.17\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.52\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.79\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e3.26\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.41\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.68\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.95\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePHE\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.57\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.38\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.41\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e3.48\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.61\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.12\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.70\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eISOLEU\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.22\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.26\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.39\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.79\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.50\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.89\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.07\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLEU\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.86\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.48\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4.85\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4.68\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.87\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.45\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e4.61\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLYS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.53\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.66\u0026thinsp;\u0026plusmn;\u0026thinsp;0.13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.59\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4.16\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.90\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.99\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.99\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"8\" nameend=\"c8\" namest=\"c1\"\u003e \u003cp\u003e\u003cb\u003eNon-essential amino acids\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGLY\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.54\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.54\u0026thinsp;\u0026plusmn;\u0026thinsp;0.17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.49\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.83\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.14\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.37\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eASP\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4.17\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e8.25\u0026thinsp;\u0026plusmn;\u0026thinsp;0.19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e7.88\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e7.70\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2.39\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2.44\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e3.70\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGLU\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e7.37\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e8.49\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e9.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e9.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.47\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2.26\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e6.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSER\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.95\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.41\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.38\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.48\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.65\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.73\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.38\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eALA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.14\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e3.21\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.54\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e3.24\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTYR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.50\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.62\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.51\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e3.09\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.19\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.25\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.20\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCYS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.09\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.23\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e3.25\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.21\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.57\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"8\"\u003eThe values were the averages of duplicate experiments.\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eDDG samples also contained most of the non-essential amino acids. Aspartic acid and glutamic acid were found as the highest amino acids in most of the DDGs regardless of the raw material. These amino acids contributed up to 7.7\u0026ndash;9.0% of dry matter in the rice DDGs (except the SLSL-R, due to the lower protein content of this sample compared to other rice DDGs) and 3.7\u0026ndash;6.1% of dry matter in the corn DDG Factory 5-CO. Alanine was found also in these DDGs at a remarkable level, especially in the rice and corn DDG, in which alanine contributed more than 3% of dry matter (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAmino acids of cassava DDG samples were lower in accordance with the lower protein ratio. Arginine, leucine, aspartic acid, glutamic acid, and alanine presented in cassava DDGs at the level above 1%. In contrast, histidine, glycine, tyrosine and cysteine amounted below the level of 0.34% (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003e3.3. Preliminarily valorization of rice and cassava DDG\u003c/h2\u003e \u003cp\u003eAt different temperatures, NaOH 0.5M (with NaHSO\u003csub\u003e3\u003c/sub\u003e 0.05M) extracted a varied amount of solid from SLSF-R. For instance, at 70\u003csup\u003eo\u003c/sup\u003eC in 2 hours, 79.31% dry weight of DDG was extracted. At lower temperatures such as 50\u003csup\u003eo\u003c/sup\u003eC and 25\u003csup\u003eo\u003c/sup\u003eC, the extraction yield was reduced to 65.66 and 56.44%, respectively (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). By adjusting pH to 4, the precipitation was generated from the extracted soluble in liquid phase. However, the precipitate amount was not in agreement with the extraction yields. For instance, the precipitate from 50\u003csup\u003eo\u003c/sup\u003eC extraction was almost the same as that from 70\u003csup\u003eo\u003c/sup\u003eC extraction (ca. 36% of the initial DDG amount). The extraction at 25\u003csup\u003eo\u003c/sup\u003eC resulted in less extracted yield and precipitation (about 27.14% of the initial DDG amount) after pH adjustment (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). The protein content in precipitates was in the range of 59.72 to 72.5% (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e) which was improved than the protein content in the initial SLSF-R (51.49%) (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eExtraction of protein from SLSF-R\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCondition of extraction\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eExtraction yield** (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eResidual solid* (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePrecipitated solid* (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eProtein content in precipitated solid (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eProtein recovery (%)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e70℃, 120 rpm, 2 hours\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e79.31\u0026thinsp;\u0026plusmn;\u0026thinsp;1.47\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e20.69\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e36.22\u0026thinsp;\u0026plusmn;\u0026thinsp;1.53\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e70.53\u0026thinsp;\u0026plusmn;\u0026thinsp;2.23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e54.82\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e50℃, 120 rpm, 2 hours\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e65.66\u0026thinsp;\u0026plusmn;\u0026thinsp;1.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e34.34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e35.90\u0026thinsp;\u0026plusmn;\u0026thinsp;0.31\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e72.50\u0026thinsp;\u0026plusmn;\u0026thinsp;2.57\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e55.85\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e25℃, 120 rpm, 2 hours\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e56.44\u0026thinsp;\u0026plusmn;\u0026thinsp;2.45\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e43.56\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e27.14\u0026thinsp;\u0026plusmn;\u0026thinsp;1.82\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e59.72\u0026thinsp;\u0026plusmn;\u0026thinsp;1.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e34.78\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"6\"\u003e* weight of solid in comparison to the initial SLS-R amount; ** calculated based on the residual solid\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe bread samples with the replacement of wheat flour by enriched protein precipitate showed differences in rheological properties compared to the control bread (without replacement) (Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e, Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). In general, the more replacement, the higher hardness, chewiness, and gumminess of bread samples was observed (Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab5\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eRheological property of bread samples with different wheat replacement ratio by enriched protein\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eParameters\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"5\" nameend=\"c6\" namest=\"c2\"\u003e \u003cp\u003eRatio of enriched protein\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5%\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e10%\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e15%\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e20%\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHardness(g)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e109.69\u0026thinsp;\u0026plusmn;\u0026thinsp;11.44\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e336.40\u0026thinsp;\u0026plusmn;\u0026thinsp;32.59\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e669.16\u0026thinsp;\u0026plusmn;\u0026thinsp;77.30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1037.06\u0026thinsp;\u0026plusmn;\u0026thinsp;80.39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1496.50\u0026thinsp;\u0026plusmn;\u0026thinsp;94.39\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCohesiveness\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.908\u0026thinsp;\u0026plusmn;\u0026thinsp;0.013\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.85\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.764\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.660\u0026thinsp;\u0026plusmn;\u0026thinsp;0.034\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.597\u0026thinsp;\u0026plusmn;\u0026thinsp;0.033\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eResilience\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.614\u0026thinsp;\u0026plusmn;\u0026thinsp;0.072\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.849\u0026thinsp;\u0026plusmn;\u0026thinsp;0.092\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.433\u0026thinsp;\u0026plusmn;\u0026thinsp;0.022\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.328\u0026thinsp;\u0026plusmn;\u0026thinsp;0.032\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.298\u0026thinsp;\u0026plusmn;\u0026thinsp;0.062\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eChewiness (N)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e98.67\u0026thinsp;\u0026plusmn;\u0026thinsp;15.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e263.48\u0026thinsp;\u0026plusmn;\u0026thinsp;28.42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e420.29\u0026thinsp;\u0026plusmn;\u0026thinsp;36.14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e612.124\u0026thinsp;\u0026plusmn;\u0026thinsp;33.33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e727.00\u0026thinsp;\u0026plusmn;\u0026thinsp;53.08\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSpringiness\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.918\u0026thinsp;\u0026plusmn;\u0026thinsp;0.011\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.949\u0026thinsp;\u0026plusmn;\u0026thinsp;0.017\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.922\u0026thinsp;\u0026plusmn;\u0026thinsp;0.023\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.871\u0026thinsp;\u0026plusmn;\u0026thinsp;0.026\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.859\u0026thinsp;\u0026plusmn;\u0026thinsp;0.022\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGumminess\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e113.70\u0026thinsp;\u0026plusmn;\u0026thinsp;12.90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e265.67\u0026thinsp;\u0026plusmn;\u0026thinsp;15.84\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e451.36\u0026thinsp;\u0026plusmn;\u0026thinsp;22.50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e703.87\u0026thinsp;\u0026plusmn;\u0026thinsp;49.53\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e845.91\u0026thinsp;\u0026plusmn;\u0026thinsp;57.02\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eFour fungal strains were used in the solid-state cultivation of cassava DDG Factory 5-CS. The protein content after 7 days of cultivation was slightly improved, ranging from 18.08\u0026ndash;20.70%, from the initial protein value in Factory 5-CS was 16.42% (Table\u0026nbsp;\u003cspan refid=\"Tab6\" class=\"InternalRef\"\u003e6\u003c/span\u003e). On the other hand, the measure of three carbohydrate-active enzymes including CMCase, xylanase and amylase showed that they were produced and accumulated in solid media at varied activity levels (Table\u0026nbsp;\u003cspan refid=\"Tab6\" class=\"InternalRef\"\u003e6\u003c/span\u003e, Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab6\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 6\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eChanges of proximate composition and enzyme accumulation of Factory 5-CS in solid state cultivation with fungal strains\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eStrain\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFiber\u003c/p\u003e \u003cp\u003e(%DM)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAsh\u003c/p\u003e \u003cp\u003e(%DM)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eProtein (%DM)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eCMCase\u003c/p\u003e \u003cp\u003e(U/g DM)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eXylanase\u003c/p\u003e \u003cp\u003e(U/g DM)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eAmylase\u003c/p\u003e \u003cp\u003e(U/g DM)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFactory 5-CS*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e33.98\u0026thinsp;\u0026plusmn;\u0026thinsp;1.28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e10.03\u0026thinsp;\u0026plusmn;\u0026thinsp;0.59\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e16.42\u0026thinsp;\u0026plusmn;\u0026thinsp;0.16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eRhizomucor miehei\u003c/em\u003e FCH 116.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e35.55\u0026thinsp;\u0026plusmn;\u0026thinsp;0.68\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e18.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e19.73\u0026thinsp;\u0026plusmn;\u0026thinsp;0.44\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e10.07\u0026thinsp;\u0026plusmn;\u0026thinsp;0.34\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eRhizopus microsporus\u003c/em\u003e LPH 084\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e35.76\u0026thinsp;\u0026plusmn;\u0026thinsp;0.90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e12.59\u0026thinsp;\u0026plusmn;\u0026thinsp;0.93\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e18.58\u0026thinsp;\u0026plusmn;\u0026thinsp;0.21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.66\u0026thinsp;\u0026plusmn;\u0026thinsp;0.39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.93\u0026thinsp;\u0026plusmn;\u0026thinsp;0.35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e14.25\u0026thinsp;\u0026plusmn;\u0026thinsp;0.73\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eRhizopus microsporus\u003c/em\u003e LPH 156\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e37.41\u0026thinsp;\u0026plusmn;\u0026thinsp;1.45\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e12.22\u0026thinsp;\u0026plusmn;\u0026thinsp;0.71\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e18.08\u0026thinsp;\u0026plusmn;\u0026thinsp;0.16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.02\u0026thinsp;\u0026plusmn;\u0026thinsp;0.76\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.39\u0026thinsp;\u0026plusmn;\u0026thinsp;0.67\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e14.44\u0026thinsp;\u0026plusmn;\u0026thinsp;0.84\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eThermothelomyces thermophilus\u003c/em\u003e FCH 112.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e36.35\u0026thinsp;\u0026plusmn;\u0026thinsp;0.66\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e15.17\u0026thinsp;\u0026plusmn;\u0026thinsp;0.60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e20.70\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e4.94\u0026thinsp;\u0026plusmn;\u0026thinsp;0.42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2.03\u0026thinsp;\u0026plusmn;\u0026thinsp;0.63\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"7\"\u003e* Sample collected in a different batch from the sample of cassava DDG in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"4. Discussion","content":"\u003cp\u003eThe wet by-product from the ethanol industry has been used for a long time for the purpose of animal feeding in Vietnam. However, the nutrient composition of this potential by-product has not been elucidated, except for little data recently published [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. In contrast, there is a huge bank of data for wheat and corn DDGS which have been published several years [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan additionalcitationids=\"CR16\" citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. In fact, rice and cassava have been known as more popular crops in Asian and African countries than maize and wheat. This study was the first systematic work focusing on the nutrient compositions of by-products from different ethanol factories in Vietnam using rice, and cassava as raw materials.\u003c/p\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003e4.1 The composition of DDG depend on bioethanol technology and raw materials\u003c/h2\u003e \u003cp\u003eIn this study, the variation of composition including crude protein, starch, fat, crude fiber and ash of 7 DDG samples collected from 5 ethanol factories was observed. The variation in composition was also proven for DDGS from corn and wheat [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. The authors showed that the types of raw material, the production technology and WDG treatment influence on the composition of DDGS, and this was an explanation for the variation in proximates observed with rice and cassava DDGs in this study. Proximate of these DDGs were different due to not only the raw material but also ethanol processing and WDG treatment (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). In general, for all materials, bioethanol manufacture starts with a liquefaction step in which milled raw material is treated at high temperature of above 100\u003csup\u003eo\u003c/sup\u003eC and supported by a heat-stable α-amylase. Among these, a high-pressure steam treatment (temperature of 105 -110\u003csup\u003eo\u003c/sup\u003eC) was applied in Factory 3 and Factory 5 to achieve the highly extensive liquefaction of starch. A saccharification step was performed either separately (as in Factory 1, Factory 2 and Factory 5) or simultaneously with fermentation (as in Factory 4 and Factory 5). Factory 5 carried out a continuous fermentation meanwhile others used batch fermentation. The SLSF sample was different from others because the liquefaction, saccharification and fermentation were simultaneously performed at 30\u003csup\u003eo\u003c/sup\u003eC with yeast in the supporting of liquefying and saccharifying enzymes.\u003c/p\u003e \u003cp\u003eMoreover, the WDG was differently harvested and treated from the factory to others. The WDG was separated by a plate-frame filter before the alcohol distillation to avoid blocking the distilling tower as at Factory 1 and Factory 2. Meanwhile, in other factories, the WDGs were harvested from the whole stillage after the alcohol distillation by a high-speed decanter (Factory 3, Factory 4) or by frame-plate filter (Factory 5). Moreover, in Factory 3, the thin stillage after the decantation was partly reused to reduce the pH of the rice mash. Interestingly, in Factory 4, spent from the cassava starch harvesting process, containing fiber up to 50% dry matter, was combined with WDG from the bioethanol process.\u003c/p\u003e \u003cp\u003eThe composition of raw materials such as white rice, cassava chips and corn has been extensively elucidated. Based on dry matter, rice (milled rice or white rice) is composed of less crude fiber (0.1\u0026ndash;0.8%) than corn (3%) or cassava (3.7- 4.0%) [\u003cspan additionalcitationids=\"CR19 CR20\" citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. On the other hand, white rice contains 7\u0026ndash;8% of crude protein, which is less than that in corn (9.42%) and higher than in cassava (1\u0026ndash;3%) [\u003cspan additionalcitationids=\"CR19 CR20\" citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. Three types of raw materials were composed of a comparable level of carbohydrates with mainly starch, i.e., 80% for rice and cassava and 72% for corn. During the bioethanol processing, the un-converted components such as crude fiber in the raw material, which are mostly left in DDG, could lead the difference in DDG composition. This explains the much higher level of protein in the rice DDG than that in the corn DDG and the cassava DDGs (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Unlike wheat/corn DDGS, even though the soluble in thin stillage was not taken into account (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e), the rice DDGs in this study contained more protein than the corn DDGS which ranged from 26\u0026ndash;32% [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e, \u003cspan additionalcitationids=\"CR23\" citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. Also, in this work the protein content in the corn DDG (Factory 5-CO) was slightly higher than the range (35.6%) (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Yeast biomass has been known as a rich source of protein which was reported in a range of 38.8\u0026ndash;70.7% dry matter [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. The amount of protein in DDGS is affected by yeast as shown in a review by Liu et al [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. In an uncertain estimation, the authors indicated that yeast contributes approximately 5.3% of the protein in DDGS [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. This number is much lower than the amount of protein observed in rice DDG in this study (50\u0026ndash;80% of dry matter). This suggests most of the protein in rice DDG was from rice material. This statement is also supported by the low protein value in cassava DDGs (13.38\u0026ndash;16.40%) (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). If protein in DDG were mainly contributed by yeast biomass, the protein content in cassava-DGG would be much higher than the observed values.\u003c/p\u003e \u003cp\u003eThe higher fiber in cassava resulted in the higher of this parameter in cassava DDGs (up to 36.5% in Factory 5-CS) in comparison to the rice DDGs (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). The high content of fiber in the cassava DDG Factory 4-CS was explained by the addition of the cassava spent by-product of starch processing which contains up to 50% of fiber. The fiber content in the corn DDG (Factory 5-CO) in this study was in agreement with the reported range for DDGS (7.22\u0026ndash;10.2%) [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e, \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe content of residual starch in DDG samples indicated the effectiveness of heat treatment, amylolytic conversion by amylases and fermentation by yeast [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. Starch content in corn DDGS ranges from 3\u0026ndash;6% [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. The starch content in DDGs in this study might be overestimated due to the analysis method using HCl hydrolysis. The reducing sugar from non-starch carbohydrates can be generated after acid hydrolysis.\u003c/p\u003e \u003cp\u003eDespite the lower ash content in cassava [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e], the ash content of cassava DDG was higher than that in rice DDG (1.44\u0026ndash;2.61%) and corn DDG (3%) (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThere was no correlation between fat content in raw material and DDG samples. For example, the fat content in rice (1\u0026ndash;2%) and cassava (0.5-1.0%) is lower than in maize (4.72%) [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e], and the fat in the rice-DDG varied from 0.69% \u0026minus;\u0026thinsp;10.41% (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). We also found in this work that the fat content in the corn DDG Factory 5-CO was 4.56%, which is half to one-third of the reported range from 10.2\u0026ndash;14.5% of DDGS [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e, \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e, \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe difference in compositions of the rice DDG samples from Factory 2-R, Factory 3-R, Factory 4-R and the SLSF-R suggests that the ethanol processing technology leads to a significant effect. Especially, the Factory 2-R and the SLSF-R used the same rice material but different process technology showing varied data in protein, starch, fiber and other components. Interestingly, the SLSF-R differs remarkably from that of Factory 2-R on protein (51.49%), fiber (18.42%) and fat (7.47%). It is likely because of the uncooked and simultaneous saccharification and fermentation in the SLSF process (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003e4.2 The potential of rice in food application and cassava DDG in animal feeding\u003c/h2\u003e \u003cp\u003eDDGS was considered not only for animal feeding but also for food application recently. This study showed that the rice DDG which contained protein up to 50\u0026ndash;80% of dry matter was highly potential for food application, especially as the source of plant protein. In comparison to soybean seed which is well-known as a main source of plant protein for food application currently, the protein in rice DDG was much higher than the protein in soybean seed (ca 40% of dry matter [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]). Moreover, the profile of amino acids in the rice DDG is balancing of essential and non-essential amino acids (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). Similarly in rice as reported previously [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e], in the rice DDG the essential amino such as isoleucine, leucine, phenylalanine, valine, threonine are also present at higher levels than others. This also means the ethanol process did not change the amino acid profile of raw material to DDG as mentioned previously [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. Lysine, an important amino acid, is considered the first limiting in cereals such as maize, cassava and even in some varieties of rice [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. In some varieties of Vietnamese rice, lysine was in the range of 3\u0026ndash;5% of total protein [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e], which is in agreement with the lysine content in rice DDGs found in this study (approximately 2.5\u0026ndash;4% of rice DDG) (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). Lysine presents in rice DDG at a level of 2.5-4 times higher than in cassava or corn DDG (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAmong rice DDGs, SLSF-R was not treated with heating, therefore protein (~\u0026thinsp;51% of dry matter) in the SLSF-R was expectedly unmodified as in other rice DDGs due to heat treatment. Protein from SLSF-R was preliminarily harvested with alkaline extraction and precipitation by pH 4.5 adjustment. The main protein fraction in rice, glutenin (amounted to 75\u0026ndash;90% of total rice protein) [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e], was reported as well-dissolved in alkaline condition [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e], therefore NaOH 0.5M was used for extraction. The protein content was increased to 70\u0026ndash;72% in resulted precipitate with a recovery yield of 54\u0026ndash;55% (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). The enriched protein precipitate from SLSF-R was applied to replace wheat flour in the bread recipe. With such high protein, the replacement of enriched-protein precipitate from SLSF-R to wheat flour can improve the protein content in bread, however, it led the reduction of gluten which is needed for the textural structure of bread, therefore the rheological properties of bread were negatively affected, especially at the high ratio of replacement [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Among these, the ratio of 5% replacement was acceptable in flavor and taste. Even though, with a such high protein content, it is highly potentially applied in many non-structural food products. Protein from rice DDG, furthermore, could be used for enzymatic hydrolysis to improve the digestibility [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e] or to obtain the bioactive peptides [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eCassava DDG contained a low level of protein but a high level of fiber. Fiber, even though does not generate energy, has to be included in the diet to maintain normal physiological functions in the digestive tract [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. Using molds for fermentation to enrich protein in cassava DDG has been reported by Mai-Dinh Vuong et al. with protein content increased from 11.96 to 15.27% using \u003cem\u003eTrichoderma harzianum\u003c/em\u003e strain [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e], similar results were observed in this study with the protein content increased from the initial value of 16.42% up to 18.08% \u0026minus;\u0026thinsp;20.7% of dry matter, depending on strains (Table\u0026nbsp;\u003cspan refid=\"Tab6\" class=\"InternalRef\"\u003e6\u003c/span\u003e). Among used fungal strains, \u003cem\u003eRhizomucor miehei\u003c/em\u003e FCH 116.3 and \u003cem\u003eThermothelomyces thermophilus\u003c/em\u003e FCH 112.2 showed better protein improvement than others. Moreover, the evaluation of carbohydrate-active enzymes showed that the strains had the ability to produce the three enzymes required in animal feed on cassava-based WDG substrate (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). The on-site production of enzymes through fermentation for animal feed production and high protein biomass generation is a promising strategy.\u003c/p\u003e \u003c/div\u003e"},{"header":"5. Conclusions","content":"\u003cp\u003eIn this study, the first systematic analysis of proximates of rice/cassava-based DDG was performed and showed the variation of proximates in DDG samples which depended on bioethanol technology, WDG treatments as well as raw materials. Among them, the rice DDGs with the high protein content and balancing amino acid profile were shown as the highly potential plant protein source for food application. Meanwhile, cassava DDG also exhibited valuable for animal feeding not only due to the high level of fiber but also the substrate source for accumulation of the animal health beneficial enzymes by solid-state fermentation with fungi.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"576\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"18.02426343154246%\" valign=\"top\"\u003e\n \u003cp\u003eCMC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"81.97573656845753%\" valign=\"top\"\u003e\n \u003cp\u003eCarboxymethylcellulose\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"18.02426343154246%\" valign=\"top\"\u003e\n \u003cp\u003eDDG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"81.97573656845753%\" valign=\"top\"\u003e\n \u003cp\u003eDistillers dried grain\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"18.02426343154246%\" valign=\"top\"\u003e\n \u003cp\u003eDDGS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"81.97573656845753%\" valign=\"top\"\u003e\n \u003cp\u003eDistillers dried grain with soluble\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"18.02426343154246%\" valign=\"top\"\u003e\n \u003cp\u003eDNS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"81.97573656845753%\" valign=\"top\"\u003e\n \u003cp\u003e3,5-dinitrosalicylic acid\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"18.02426343154246%\" valign=\"top\"\u003e\n \u003cp\u003ePDA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"81.97573656845753%\" valign=\"top\"\u003e\n \u003cp\u003ePotato Dextrose Agar\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"18.02426343154246%\" valign=\"top\"\u003e\n \u003cp\u003eSLSF\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"81.97573656845753%\" valign=\"top\"\u003e\n \u003cp\u003eSimultaneous liquefaction saccharification and fermentation process\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"18.02426343154246%\" valign=\"top\"\u003e\n \u003cp\u003eWDG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"81.97573656845753%\" valign=\"top\"\u003e\n \u003cp\u003eWet distillers grain\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"18.02426343154246%\" valign=\"top\"\u003e\n \u003cp\u003eWDGS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"81.97573656845753%\" valign=\"top\"\u003e\n \u003cp\u003eWet distiller\u0026rsquo;s grains with soluble\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eConflict of interest\u0026nbsp;\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no conflict of interest.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis work was supported by project NVQG2020/ĐT.01, Ministry of Science and Technology, Vietnam\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTien-Thanh Nguyen performed the experiments and wrote the manuscripts.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eBach Cao-Xuan performed experiments and drafted partly the manuscripts.\u003c/p\u003e\n\u003cp\u003ePham Kim Dang, Thuy Nguyen-Thanh, Tien Cuong Nguyen revised partly the manuscripts.\u003c/p\u003e\n\u003cp\u003eNguyen Gia Long, Mai Dinh Vuong performed the experiments.\u003c/p\u003e\n\u003cp\u003eChu-Ky Son, Vu Nguyen Thanh supervised, finalized, and submitted.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData availability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe datasets are available from the corresponding author on reasonable request.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eMonceaux, D. and D. Kuehner: Dryhouse technologies and DDGS production. Lallemand ethanol technology and Nottingham University Press, (2009).\u003c/li\u003e\n\u003cli\u003eLiu, K.: Chemical composition of distillers grains, a review\u003cem\u003e.\u003c/em\u003e J. Agri. Food Chem., 59(5), 1508-1526 (2011).\u003c/li\u003e\n\u003cli\u003eLi, X., C. Wang, and P.G. Krishnan: Effects of corn distillers dried grains on dough properties and quality of Chinese steamed bread\u003cem\u003e.\u003c/em\u003e Food Sci. Nutr., 8(8), 3999-4008 (2020).\u003c/li\u003e\n\u003cli\u003ePourafshar, S., K.A. Rosentrater, and P.G. Krishnan: Changes in chemical and physical properties of Latin American wheat flour based tortillas substituted with different levels of distillers dried grains with solubles (DDGS)\u003cem\u003e.\u003c/em\u003e J. Food Sci. Technol., 52(8), 5243-5249 (2015).\u003c/li\u003e\n\u003cli\u003ePourafshar, S., K.A. Rosentrater, and P.G. Krishnan: Production of Barbari bread (traditional Iranian bread) using different levels of distillers dried grains with solubles (DDGS) and sodium stearoyl lactate (SSL)\u003cem\u003e.\u003c/em\u003e Foods, 7(3), 31 (2018).\u003c/li\u003e\n\u003cli\u003eRosentrater, K.: Incorporating distillers grains in food products\u003cem\u003e.\u003c/em\u003e Cereal Foods World - CFW, 51, 52-60 (2006).\u003c/li\u003e\n\u003cli\u003eSingha, P., K. Muthukumarappan, and P. Krishnan: Influence of processing conditions on apparent viscosity and system parameters during extrusion of distiller\u0026apos;s dried grains-based snacks\u003cem\u003e.\u003c/em\u003e Food Sci. Nutr., 6(1), 101-110 (2018).\u003c/li\u003e\n\u003cli\u003eMinistry-of-Trade-and-Industry-of-Vietnam: Targets of the scheme to develop of beverage in Vietnam to 2025 and the vision to 2035 (Decision No 3690/QĐ-BCT). 2016: 12/06/2016.\u003c/li\u003e\n\u003cli\u003eXuan, D.N.N., M.L. Huu, and U. Peter: Tropical fibre sources for pigs\u0026mdash;digestibility, digesta retention and estimation of fibre digestibility in vitro\u003cem\u003e.\u003c/em\u003e Anim. Feed Sci. Technol., 102(1\u0026ndash;4), 109-124 (2002).\u003c/li\u003e\n\u003cli\u003eTaranu, I., et al.: Rice and cassava distillers dried grains in Vietnam: Nutritional values and effects of their dietary inclusion on blood chemical parameters and immune Responses of Growing Pigs\u003cem\u003e.\u003c/em\u003e Waste and Biomass Valorization, 10(11), 3373-3382 (2019).\u003c/li\u003e\n\u003cli\u003eDinh Vuong, M., et al.: Protein enrichment of cassava-based dried distiller\u0026rsquo;s grain by solid state fermentation using \u003cem\u003eTrichoderma harzianum\u003c/em\u003e and \u003cem\u003eYarrowia lipolytica\u003c/em\u003e for feed ingredients\u003cem\u003e.\u003c/em\u003e Waste and Biomass Valorization, 12, 3875-3888 (2021).\u003c/li\u003e\n\u003cli\u003eMiller, G.L.: Use of dinitrosalicylic acid reagent for determination of reducing sugar\u003cem\u003e.\u003c/em\u003e Analytical Chemistry, 31(3), 426-428 (1959).\u003c/li\u003e\n\u003cli\u003eDo, T.-T.-H., et al.: Extraction and hydrolysis of protein from industrial rice-based dried distiller\u0026rsquo;s grain toward food application (origin text in Vietnamese)\u003cem\u003e.\u003c/em\u003e J. Vietnam Agri. Sci. Technol., 01(122), 106 - 112 (2021).\u003c/li\u003e\n\u003cli\u003eThanh, V.N., et al.: Surveying of acid-tolerant thermophilic lignocellulolytic fungi in Vietnam reveals surprisingly high genetic diversity\u003cem\u003e.\u003c/em\u003e Scientific Reports, 9(1), 3674 (2019).\u003c/li\u003e\n\u003cli\u003eAlag\u0026oacute;n, G., et al.: Nutritive value of distillers dried grains with solubles from barley, corn and wheat for growing rabbits\u003cem\u003e.\u003c/em\u003e Anim. Feed Sci. Technol., 222, 217-226 (2016).\u003c/li\u003e\n\u003cli\u003eBelyea, R.L., et al.: Sources of variation in composition of DDGS\u003cem\u003e.\u003c/em\u003e Anim. Feed Sci. Technol., 159(3), 122-130 (2010).\u003c/li\u003e\n\u003cli\u003eBelyea, R.L., K.D. Rausch, and M.E. Tumbleson: Composition of corn and distillers dried grains with solubles from dry grind ethanol processing\u003cem\u003e.\u003c/em\u003e Bioresour. Technol., 94(3), 293-298 (2004).\u003c/li\u003e\n\u003cli\u003eMontagnac, J.A., C.R. Davis, and S.A. Tanumihardjo: Nutritional value of cassava for use as a staple food and recent advances for improvement\u003cem\u003e.\u003c/em\u003e Compr. Rev. Food Sci. F., 8(3), 181-194 (2009).\u003c/li\u003e\n\u003cli\u003eNuss, E.T. and S.A. Tanumihardjo: Maize: A paramount staple crop in the context of global nutrition\u003cem\u003e.\u003c/em\u003e Compr. Rev. Food Sci. F., 9(4), 417-436 (2010).\u003c/li\u003e\n\u003cli\u003eMorgan, N.K. and M. Choct: Cassava: Nutrient composition and nutritive value in poultry diets\u003cem\u003e.\u003c/em\u003e Anim. Nutri., 2(4), 253-261 (2016).\u003c/li\u003e\n\u003cli\u003eZhou, Z., et al.: Composition and functional properties of rice\u003cem\u003e.\u003c/em\u003e Inter. J. Food Sci. Technol., 37(8), 849-868 (2002).\u003c/li\u003e\n\u003cli\u003eCromwell, G.L., K.L. Herkelman, and T.S. Stahly: Physical, chemical, and nutritional characteristics of distillers dried grains with solubles for chicks and pigs\u003cem\u003e.\u003c/em\u003e J. Anim. Sci., 71(3), 679-686 (1993).\u003c/li\u003e\n\u003cli\u003eKim, Y., et al.: Composition of corn dry-grind ethanol by-products: DDGS, wet cake, and thin stillage\u003cem\u003e.\u003c/em\u003e Bioresour. Technol., 99(12), 5165-5176 (2008).\u003c/li\u003e\n\u003cli\u003eSpiehs, M.J., M.H. Whitney, and G.C. Shurson: Nutrient database for distiller\u0026apos;s dried grains with solubles produced from new ethanol plants in Minnesota and South Dakota\u003cem\u003e.\u003c/em\u003e J. Anim. Sci. , 80(10), 2639-2645 (2002).\u003c/li\u003e\n\u003cli\u003eMartini, A.E.V., M.W. Miller, and A. Martini: Amino acid composition of whole cells of different yeasts\u003cem\u003e.\u003c/em\u003e J. Agri. Food Chem., 27(5), 982-984 (1979).\u003c/li\u003e\n\u003cli\u003eMolinari, M.D., et al.: Exploring the proteomic profile of soybean bran: Unlocking the potential for improving protein quality and quantity\u003cem\u003e.\u003c/em\u003e Plants, 12(14)(2023).\u003c/li\u003e\n\u003cli\u003eKhoi, B.H., et al.: The protein and the amino acid composition of some rice and maize varieties grown in North Vietnam\u003cem\u003e.\u003c/em\u003e J. Sci. Food Agri., 39(2), 137-143 (1987).\u003c/li\u003e\n\u003cli\u003eTorbatinejad, N.M., S.M. Rutherfurd, and P.J. Moughan: Total and reactive lysine contents in selected cereal-based food products\u003cem\u003e.\u003c/em\u003e J. Agri. Food Chem., 53(11), 4454-4458 (2005).\u003c/li\u003e\n\u003cli\u003eKawakatsu, T. and F. Takaiwa: 4 - Rice proteins and essential amino acids. Rice (Fourth Edition), ed. J. Bao. AACC International Press, (2019).\u003c/li\u003e\n\u003cli\u003eCookman, D.J. and C.E. Glatz: Extraction of protein from distiller\u0026rsquo;s grain\u003cem\u003e.\u003c/em\u003e Bioresource Technology, 100(6), 2012-2017 (2009).\u003c/li\u003e\n\u003cli\u003eSelamassakul, O., et al.: Bioactive peptides from brown rice protein hydrolyzed by bromelain: Relationship between biofunctional activities and flavor characteristics\u003cem\u003e.\u003c/em\u003e J Food Sci, 85(3), 707-717 (2020).\u003c/li\u003e\n\u003cli\u003eLindberg, J.E.: Fiber effects in nutrition and gut health in pigs\u003cem\u003e.\u003c/em\u003e J. Anim. Sci. Biotechnol., 5(1), 15 (2014).\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"rice, cassava, bioethanol by-product, proximates, non-animal protein, food application","lastPublishedDoi":"10.21203/rs.3.rs-3171967/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-3171967/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eSeveral bioethanol plants in Vietnam using rice or cassava as raw materialsgenerate a large number of by-products in the form of wet distillers spent grain, which have been underutilized with simple treatment for animal feeding. In this work, the biochemical and nutritional values such as protein (amino acid profile), lipid, fiber, ash, starch, calcium and phosphorous of dried distillers’ grain (DDG) collected from different bioethanolfactories in Vietnam were assessed. The DDG samples were shown to vary in nutrient compositions which depended either on raw materials or ethanol processing technology. Among, rice DDG was shown as the most nutritively valuable with very high protein content (55-80% of dry matter) and appropriate amino acid profile whereas cassava DDG was characterized by a high fiber content and a low protein content (13-16%). The protein in rice based DDG could be enriched by extraction/precipitation and applied for food products. The obtained data suggest that the by-products from rice-based bioethanol are very potential and promise to be used efficiently as ingredients not only for the animal feed industry but also for the food industry.\u003c/p\u003e","manuscriptTitle":"Rice-Based Distillers’ Dried Grains from Bio-Ethanol Production, a Potential Source of Protein for Food Application","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-03-05 15:04:47","doi":"10.21203/rs.3.rs-3171967/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"666381f5-f0fe-4dcc-be7a-a0fcce7b110e","owner":[],"postedDate":"March 5th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-03-31T15:08:40+00:00","versionOfRecord":[],"versionCreatedAt":"2024-03-05 15:04:47","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-3171967","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-3171967","identity":"rs-3171967","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}