{"paper_id":"bc2b6958-27f7-4e5b-a475-8136b0757bf9","body_text":"Production of New Nano-Bacterial Cellulose with Lactobacillus rhamnosus by Using Whey Waste as Substrate with Optimization Taguchi Method, which has the potential to be used in many biomedical products | 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 Production of New Nano-Bacterial Cellulose with Lactobacillus rhamnosus by Using Whey Waste as Substrate with Optimization Taguchi Method, which has the potential to be used in many biomedical products Aytül Bayraktar, Cansu Gürsoy This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3828016/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 Whey waste, which has a negative impact on the environment, is an important component with high organic content. The fact that it contains lactose, a fermentable sugar, is a suitable substrate for the formation of natural nano-cellulose. Bacterial nano-cellulose (BNC), a type of natural cellulose polymer synthesized by some microorganisms, has been reported to be a promising natural biomedical material due to its distinctive feature, including its unique fibril nanostructure, high water holding capacity, crystallinity, high chemical purity, fine wet mechanical property. In this study, new BNC production was realized for the first time by using Lactobacillus rhamnosus bacteria and whey as organic substrate. Optimum condition was determined by Taguchi method under the following condition; pH (5-6), organic source concentration (25-100 % g/L), active culture (10-30 % g/L), incubation period (8-12 day). Whereas Taguchi method was highest performed at at pH 5.5, organic source concentration 25 % g/L, active culture 30 % g/L, incubation period 8 days with 5.41 g BNC yield. Effects of organic source concentration found as decisive factor on Lactobacillus rhamnosus BNC yield with 95% confidence interval. Field emission scanning electron microscopy (FESEM), fourier transform infrared spectroscopy (FTIR), differential / thermogravimetric thermal analysis (DTG/TG) were utilised to evaluate the structure and characterization of BNC. BNC production by Lactobacillus rhamnosus , with its biocompatible and biodegradable properties, environmentally friendly and low-cost nanomaterials have been produced with the potential to be used in many biomedical applications such as wound dressing and drug coating material. Lactobacillus rhamnosus bacterial cellulose taguchi design whey waste Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 1. Introduction Bacterial cellulose (BNC) known as a nanomaterial and could be produced by some bacteria such as Gluconacetobacter, Acetobacter spp., Rhizobium spp., Agrobacterium spp., Alcaligenes spp., Achromobacter and Sarcina [ 1 , 2 , 3 , 4 ]. The BNC biosynthesis is a regulated process including enzymes, catalytic complexes reactions and can be investigated wih two steps such as intracellular producted of 1,4-b-glucan chains and crystallization of cellulose chains [ 3 ]. BNC is native cellulose which has high crystallinity, water capacity and strength also biocompatible with plant cellulose [ 4 ]. Also it produces much thinner 3D dimensional nanoporous networks with approximately 30–50 nm than plant cellulose [ 5 ]. It has the same molecular formula as BNC and plant celluloses (C6H10O5)n but BNC doesn’t contain lignin, pectin, and hemicelluloses; thus BNC purification doesn’t require harsh chemicals and also purification is easy to apply [ 6 ]. Having high surface area and high porosity, BNC is a material that shows high performance with other compounds in physical interaction [ 3 , 7 ]. This article presents an alternative strategy for the future using whey, which is waste, as well as the role of raw material for low-cost production of bacterial cellulose. Whey obtained from whey as a by-product of the dairy industry is considered a pollutant due to its high biological oxygen demand and high cost of evaluation [ 8 ]. Besides, it has rich nutrient content [ 9 ]. Lactic acid bacteria are generally used in dairy products, as pure culture. LABs are organisms that can be isolated from natural environments such as most people, animals and plants, adapted to a specific specific environment and are found in almost natural habitats. Lactobacillus rhamnosus strains with probiotic properties have begun to be used in new types of yoghurt [ 10 ]. With the increasing demand of consumers for probiotic products, the production of lactic acid bacteria in the industrial sector has gained importance [ 11 ]. In addition to the effects of other factors in obtaining bacterial cellulose, it is crucial to determine the optimum values of some factors such as fermentation time and pH that maximize the bacterial cellulose yield [ 12 ]. In the present study, bacterial nano-cellulose (BNC) production was optimized based from different concentration substrate, as well as temperature, pH, and incubation time. According to the objectives of this study were determined as identifying whether the bacteria is producing Lactobacillus rhamnosus BNC for the first time in literature, identify an optimal media formulation for improved BNC production and characterization BNC production using fourier transform infrared spectroscopy (FT-IR) and field emission-scanning electron microscopy (FE-SEM). The substrate and process variables needed are optimized to ensure maximum BNC production at low costs. In optimization studies, while all other parameters are kept constant, one parameter is varied. In this case, it is not possible to evaluate the interaction of the parameters with each other. For this reason, Taguchi's method was used in this study to determine the effects of the factors affecting the BNC production process. Also, the evaluation of lactose in whey, which is considered as waste water with a complex organic matter content, it provides the advantage of environmentally friendly application to this study. 2. Materials and Methods 2.1. Procurement of whey Cheese whey was used as organic source and kindly obtained from Isparta Ünsüt Company (Turkey). Under aseptic conditions, samples were collected and stored at 4°C in the laboratory. Then samples were stored at -20°C before the analysis. It contains 60–62% lactose. The pH of the whey was adjusted to 6. Protein precipitation was induced by autoclaving from the whey at 121 oC for 20 min and after removed by filtration. 2.2. Microorganism and Culture Media Lyophilized culture of Lactobacillus rhamnasus (DSM, DELVO-ADD 100R, Netherlands) was used in all experiments. The stock culture of bacterial strain Lactobacillus rhamnasus was stored at − 80°C in MRS medium as 15% glycerol. Precultures were per liter prepared in the 100 mL MRS culture medium containing, 1g peptone, 2g glucose, 1g beef extract, 0.2g ammonium citrate, 0.01 g magnesium sulphate, 0.5g sodium acetate, 0.005 g manganese sulphate, and 0.2 g di-potassium phosphate. A 5 mL of this culture was used for further inoculation in the 100 mL MRS production media. The media contains molasses, yeast extract, MnSO4.4H2O, Tween 80, urea, and CaCO3. Table 1 shows the design of the experiments on the set of orthogonal arrays with respect to the selected factors and levels designed with using the taguchi method. The effect of organic sources concentration, active culture, pH and incubation time on the production of bacterial cellulose was investigated using the bacteria Lactobacillus rhamnasus. Table 1 Design of experimental factors and levels Factors Level 1 Level 2 Level 3 pH 5.0 5.5 6.0 Organic source concentration 50% Whey 25% Whey 75% Whey Amount of active culture 10% 20% 30% Incubation time (day) 8 10 12 2.3. Production of BNC Fermentations were performed as batch fermentations with Lactobacillus rhamnasus . It was cultivated in 100 mL liquid medium in 250 mL erlenmeyer flasks. Neutralizing agents were added into fermentation medium to control the pH during the fermentation. Calcium carbonate was used as neutralizing agent due to using widely for flask investigations and bioreactor processes [ 13 ]. Also, in this study, CaCO3 (60% (w/v) of the initial lactose concentration) was used as a neutralizing agent. The medium was inoculated with about (10% w/v) inoculum of bacteria. Fermentations were carried out with a temperature controlled orbital shaker at a stirring speed of 150 rpm and fermentation temperatures were chosen as the optimum growth temperatures given in Table 1 . All experiments were carried out in duplicate. 2.4. Purification of BNC pellicles After BNC was produced at 30 oC for 8 days in optimum conditions, the BNC pellicles were harvested by discarding the medium and washing with large amounts of nanopure water. The pellicles were then rinsed three times with 0.1M NaOH (Sigma Aldrich, Germany) at 60°C for 4 hours to remove sticky bacteria. Finally, the samples were repeatedly washed until the pH was below 7.0 with copious amounts of distilled water. Until a constant weight was reached the wet cellulose pellets were dried at 60°C, then the concentration was determined as g / L (mass (g) BNC / volume of culture medium (L). For further instrumental analysis isolated pellicles were used. 2.5. Characterizasyon of BNC The The microbial cellulose obtained as a result of incubation was ground in a laboratory mill (Retsch GE) after washing and drying, and particle size was obtained as 40 mesh. Viscometer measurements were performed in cuvettes with 35 ml chambers. 3.5 g of cellulose was added and the volume of the solution was completed to 35 ml with distilled water. The viscosimetric values of this 10% suspension (Vibro Viscometer SU-10) were measured in mPa.s by taking a reading every 20 seconds in the 20–21°C range, 30 Hertz frequency, constant vibration. The shaking method was used in order to measure the water holding capacity of the samples [ 14 ]. Two shakes were made and weighed to remove excess water from the surface of the samples. The samples were dried at the room temperature for 48 hours and then dried at 60°C for 12 hours to completely remove the water content. The water holding capacity was calculated using the following method: $$\\text{W}\\text{a}\\text{t}\\text{e}\\text{r} \\text{H}\\text{o}\\text{l}\\text{d}\\text{i}\\text{n}\\text{g} \\text{C}\\text{a}\\text{p}\\text{a}\\text{c}\\text{i}\\text{t}\\text{y}=\\frac{\\text{M}\\text{a}\\text{s}\\text{s} \\text{o}\\text{f} \\text{w}\\text{a}\\text{t}\\text{e}\\text{r} \\text{r}\\text{e}\\text{m}\\text{o}\\text{v}\\text{e}\\text{d} \\text{d}\\text{u}\\text{r}\\text{i}\\text{n}\\text{g} \\text{d}\\text{r}\\text{y}\\text{i}\\text{n}\\text{g} \\left(\\text{g}\\right)}{\\text{D}\\text{r}\\text{y} \\text{w}\\text{e}\\text{i}\\text{g}\\text{h}\\text{t} \\text{o}\\text{f} \\text{b}\\text{a}\\text{c}\\text{t}\\text{e}\\text{r}\\text{i}\\text{a}\\text{l} \\text{n}\\text{a}\\text{n}\\text{o}-\\text{c}\\text{e}\\text{l}\\text{l}\\text{u}\\text{l}\\text{o}\\text{s}\\text{e} \\left(\\text{g}\\right)}$$ 1 Thermal gravimetric analysis was performed to measure the weight loss and BNC thermal stability of the analyzed material when the temperature increased linearly [ 15 ]. Thermal degradation behavior of bacterial cellulose was investigated at 0-900 oC using the TGA / TDA device (Perkin Elmer diamond, USA). Percent and derivative weight loss were recorded against temperature for all samples. Solubility characteristics of microbial cellulose were determined using various solvents. The solvents used for solubility tests are toluene, acetone, chloroform, benzene, carbon tetrachloride, and dimethylformamide. In the experiment, 0.1 g of sample was placed in 10 mL of solvent and the tubes were rinsed at room temperature for 24 h to determine their solubility in solvents [ 16 ]. To determine the elemental contents of the BNC samples obtained as a result of experimental studies, FT-IR Spectroscopy was evaluated. It has been concluded whether pure cellulose is obtained from the numerical percentage of the elemental C content in the FT-IR bands of the cellulose sample. FT-IR analysis was carried out in Süleyman Demirel University Experimental and Observational Student Research and Application Center Laboratory. FT-IR spectra of the BNC pellicle were obtained with Perkin-Elmer FT-IR spectrophotometer (Norwalk, USA) in diffuse reflectance mode at a resolution of 4 particles/cm in KBr pellets. Fe-SEM was used to determine the surface structure of BNC cellulose samples obtained after drying for 10 days at 28°C in the liquid medium. Fe-SEM analysis was carried out at Isparta Süleyman Demirel University Energy Research and Research Application Center. Microstructures and morphology of samples were evaluated by examining images in three dimensions with TEM.. 2.6. Taguchi Methodology Taguchi is one of the impressive ways to design an experiment to optimize the process. Unlike traditional full factor research, it can be used as a powerful tool to optimize performance characteristics of process parameters. The Taguchi method uses orthogonal array design (OAD) while designing less number of experiments with a large number of parameters [ 17 ]. The signal / noise (S / N) ratio enables the quality characteristics of the process to be measured precisely at different process levels of controllable and uncontrollable factors. In addition, it makes it possible to minimize the negative effects of uncontrollable factors on the process. It is possible to provide the optimum parameters of each parameter in the process by determining the S / N ratios. The measurement of the quality of the results is measured by the signal-to-noise (S / N) ratio with three different characteristics of the target values: “larger is better”, “smaller is better” or “nominal is better” [ 18 ]. In this study, target values of “smaller is better” was chosen. Because it is aimed to determine the highest production at the lowest concentrations of the parameters. Thus, it is determined how the effects of the selected parameters at the lowest levels affect the BNC production efficiency. In this study, interactions between factors were not taken into account and the main effects of four parameters affecting the process were evaluated. 2.7. Design of Experiments and Choice of Factors In this study the Taguchi method was used as an experimental design and analysis method. The L9 orthogonal arrays improved by Taguchi method. The parameters determined by examining the literature information and their levels are given in Table 2 . BNC production was determined by optimization conditions, four different variables were selected as pH, carbon sources concentrations, amount of active culture conditions, and incubation time. Also,3 different levels were determined for each parameter. The main steps of this method was created firstly determination of factors, interactions and the levels of each factor. Minitab 16 software (Minitab Inc., USA) was used to create the experimental design and evaluate the data. To determine the optimization of the BNC manufacturing process, how various factors influence the process individually as well as how they contribute to the complexity of the entire process and their multiple interactions were also evaluated. By measuring the predicted interactions of different factors (based on the severity index (SI)), the effects of four different factors used in the experiment set at various levels were determined. Data obtained from Taguchi design experiment and all parameters were subjected to analysis of variance (ANOVA) for the significant difference between BNC production efficiency. P values less than 0.05 were considered significant. All statistical analyzes were performed using MINITAB v.16 (Minitab Inc., USA) statistical software package. Table 2 Taguchi L9 orthogonal experimental design Experiment No pH Organic source concentration (%) Active culture (%) Incubation time (day) 1 5.0 25 10 8 2 5.0 50 20 10 3 5.0 75 30 12 4 5.5 25 20 12 5 5.5 50 30 8 6 5.5 75 10 10 7 6.0 25 30 10 8 6.0 50 10 12 9 6.0 75 20 8 3. Results and discussion 3.1. Taguchi Method In In this study, three separate levels were defined for each of the BNC production process parameters and 81 separate experiments were required to complete the process. Instead, experiments were carried out by applying the L9 orthogonal array scheme, which requires nine experiments with the taguchi method. Experimental results were evaluated according to the Taguchi OA L9 method. Figure 1 . shows main effect plots for S/N ratio for BNC production in Lactobacillus rhamnasus with different variables, including pH (5–6), organic source concentration (25–100), active culture (10–30), incubation period (8–12). The main effect graphs showing the signal-to-noise (S / N) ratio describing the scattering around a target value were obtained by using Taguchi method. Noise factors are seen as the reason for the variability of the answers examined for the target value. S / N ratio measures the performance level and the effects of noise factors on performance. The variance decomposition is obtained for the relative effects of different production parameters on BNC productivity yield, which is called variance analysis (ANOVA) [ 19 ]. The ANOVA was investigated to find the effects of process parameters on BNC production. Different levels of BNC production were evaluated the effect of individual parameters for the conversion. According to Taguchi Method, the optimum Lactobacillus rhamnasus production conditions was obtained at pH 5.5, organic source concentration 25%, active culture 30%, incubation period 8 days. In the case of the design with the Taguchi method, when the experimental data and the estimation data were examined, 4.3 g BNC yield was obtained at pH 5.8 and the organic source concentration was 40%. While, the active culture amount was 10% at pH 5.5, 5 g BNC yield was obtained. If the organic source concentration was 70% and the active culture amount was 25%, the amount of BNC yield was 2 g. When the organic source concentration was 70% and the incubation time selected as 10 days, the amount of BNC yield was 2 g. 0.2 g BNC yield was obtained with 20%amount of active culture and 10 days incubation time. The Taguchi method based on OA detects optimum levels of process parameters by reducing variance for experiments. The results were then converted into S / N ratio data. S / N ratio of Taguchi method was used for data analysis and estimation of optimum parameters. Taguchi method factor design gives good results in the effect studies of various combinations. Figure 2 . shows the effects of four parameters such as pH, organic source concentration, incubation time and active culture concentration on BNC yield using Taguchi experimental design. pH and incubation times have been shown to positive effect on production efficiency by affecting BNC production. The effect of organic source concentration varies depending on the amount of active culture. Increasing in the amount of active culture and increasing the number of individuals in the medium caused a decrease in BNC efficiency due to the insufficient organic source in the medium. The relative interactions of the factors on the BNC process performance are shown in Table 3 . Table 3 shows that tested model has shown that model effectiveness was found as R2 = 96.22%. Analysis of variance (ANOVA) determined the significance of each independent variable which was presented in (Table 4 ). With 95% confidence interval, all parameters have marginally significant effects (P > 0.05). Table 3 General Linear Model: BNC value versus pH; organic source, active culture analysis of variance for BNC value, using sequential SS for tests. Source DF Seq SS Adj SS Seq MS F P pH 2 4.6769 4.6769 2.3384 2.49 0.409 Organic source concentration 2 9.1785 9.1785 4.5892 4.89 0.305 Active culture 2 4.4193 4.4193 2.2096 2.35 0.419 Incubation period 1 5.5873 5.5873 5.5873 5.95 0.248 Error 1 0.9384 0.9384 0.9384 Total 8 24.8004 S = 0.968736 R-Sq = 96.22% R-Sq(adj) = 69.73% Table 4 Analysis of variance (ANOVA) evaluation on BNC yield. Experiment No pH Organic Source Concentration (g/L%) Active Culture (g/L%) Incubation time (day) BNC value (g) SNRA1 1 5.0 25 10 8 0.06 24.4370 2 5.0 50 20 10 1.13 -1.0616 3 5.0 75 30 12 0.67 3.4785 4 5.5 25 20 12 0.00 * 5 5.5 50 30 8 0.23 12.7654 6 5.5 75 10 10 0.00 * 7 6.0 25 30 10 0.00 * 8 6.0 50 10 12 5.41 -14.6639 9 6.0 75 20 8 0.00 * 3.2. FTIR Figure 3 indicates FTIR spectra of L. rhamnosus cellulose. According to the analysis results, the broad band intramolecular and intermolecular hydrogen bonding -OH stretching at 3430 cm-1 observed in the FTIR spectrum of the BNC sample obtained from L. rhamnosus. Other researchers also found major stretching peak associated with –OH groups on the glucose rings and water molecules take place between 3300 cm-1 and 3600 cm-1 [ 20 , 21 ]. Ali et al., (2001) found that the C-H stretching vibration bands of -CH3 and –CH2 groups at 2926 cm-1 whereas bending bands of these groups at 1458 and 1562 cm-1. Also, in our study the intensities of bands at 2926 cm-1 and 1628 cm-1 were observed for L. rhamnosus cellulose [ 22 ]. FTIR results showed that intensities of bands at 2926 cm-1 and 1628 cm-1 were monitored for L. rhamnosus cellulose. This peak intensity is indicative of the existence of more aliphatic carbon groups (-CH2-). These spectra of standard and produced cellulose showed that these compounds obtained from different media. The band at 1628 cm-1 is associated with water in cellulose and probably some hemicelluloses [ 23 , 24 , 25 ]. In a study with microcrystalline cellulose and bamboo fibers, the H-bound OH groups were seen at 3340–3412 cm-1 wave length, and the broader band gap CH tension was similaries to our study (2968 − 2900 cm-1) [ 26 ]. 3.3. Water Release Feature The BNC yield was found as 60.4 g/g for L. rhamnosus bacterium. Water release rate of cellulose produced by L. rhamnosus bacteria was determined as 64.4%. The water release rate of celluloses during the drying period was determined as 28.4%. The water release process of the cellulose produced by the L. rhamnosus bacterium at the end of the 7 hours. The WHC of the BNCs produced in this study was in a suitable agreement with literature-cited publications [ 27 ]. The viscosity value of L. rhamnosus BS solution was found as 36 mPa.s at 0. Seconds (Fig. 4 .). A linear decrease in viscosity values was observed by dilution of BS solution. It was determined that the viscosity decreased from 36 mPa.s to 31.2 mPa.s within 3 minute. The WHC (water holding capacity) was considered as an most important physical characteristics due to biomedical application of BNC. The variations between the WHC were related with porosity and surface area. The greater surface area and larger pore size allow more water to penetrate and indwell in the BNC matrix. 3.4. Thermal properties Figure 5 . shows the thermogravimetric /differential thermal analysis (TG/DTG) curves of L. rhamnosus BNC sample. The thermal degradation behaviors of cellulose produced by L. rhamnosus were investigated and the temperatures of decomposition were determined. The graphic in Fig. 5 shows that thermal degradation occurs in two different stages. Mangut et al. (2006) found that first degradation step was started at 250°C and ended about 430°C and peak was obtained at 350°C. It means that it is cellulosic materials. At this temperature, simple structures such as CO, CO2, H2O, CH4, CH3OH are being removed by decarboxylation and demethoxylation. The second decay was observed at 780°C [ 28 ]. In this step, the aromatic components are probably separated more slowly. Due to the pyrolysis and decomposition of the cellulose concentration it is seen that a slower weight loss from 350°C to 700°C to form a crosslinking ring structure. Many studies have shown the behavior of cellulose in inorganic and organic solvents. Although the ability to dissolve in various solvents is of importance for the usage of BNC in different industry, it is also important to reveal the effects of organism differences on the cellulosic composition. Toluene, acetone, chloroform, benzene, carbon tetrachloride and dimethylformamide solvents were used for solubility tests of the cellulose samples produced by L rhamnasus bacterium. In the experiment, 0.1 g sample was placed in 10 ml of solvent and the tubes were shaken for 24 hours at room temperature, and their dissolution ability in the solvents was determined. As a result of the solubility test, it was determined that BNC was dissolved only in toluene solvent in 80oC water bath. Solubility, breaking hydrogen bonds between polymer chains and increasing the accessibility to the intercrystalline structure can be explained. The DTG curve for L. rhamnosus BNC sample showed that higher temperatures was shifted by increasing crystallite size. This behavior suggests that higher crystallite size shows higher thermal stability. Kim et al. (2010) also found that an increase in the crystallite size promoted by higher thermal stability. Therefore, BNC sample has probably higher thermal stability because it has higher amount of hydrogen bonds between cellulose chains and leads to more ordered and packed cellulose regions [ 29 ]. High crystallite BNC can possibly turn into a increasing of the thermal decomposition temperature of cellulose. The intensity of L. Rhamnosus BNC, which is believed to be rich in aromatic structures, supports the FTIR analysis results. Figure 6 . shows the cellulosic structure produced by FE-SEM electron microscopy images of L. rhamnosus bacteria with x400 and x24000 magnification, respectively. Figure 6 . shows that it was determined that the cellulosic fibers formed a tubular network structure. The cellulose fibers in these images were reinforced by determining their chemical and physical properties. 4. Conclusion In this study, the production of BNC was optimized using different variables such as pH, organic source concentration, active culture concentration, and incubation period by Taguchi method. Characterization tests showed that the produced BNC revealed a strong network of nano fibrils. In addition, it does not involve toxic and any kind of hazardous materials in producing BNC, which is excellent and suitable for safe environments such as medical, a packing material in fields such as wound healing, drug coating, vein production, food industry and cosmetics applications. In conclusion, BNC produced from L. rhamnosus could play an crucial and environmentally friendly role in diverse medical and industrial applications as another way to cellulose obtained from other sources, such as microorganism and plants. Declarations Ethical approval is not required. There is no funding received. The dataset was not used. Author Contribution A.B. and C.D. wrote the body of the manuscript and all authors reviewed the manuscript. References Iqbal H.M.N., Kyazze G., Locke I.C., Tron T. & Keshavarz T., Development of bio-composites with novel characteristics: Evaluation of phenol-induced antibacterial, biocompatible and biodegradable behaviours, Carbohydrate Polymers,2015, 131, 197–207. Gallegos A.M.A., Herrera Carrera S., Parra R., Keshavarz T. & Iqbal H.M.N., Bacterial Cellulose: A Sustainable Source to Develop Value-Added Products – A Review. BioResources,2016, 11(2), 5641–5655, DOI: 10.15376/biores.11.2.Gallegos . Azeredo H.M.C., Barud H., Farinas C.S., Vasconcellos V.M. & Claro A.M., Bacterial Cellulose as a Raw Material for Food and Food Packaging Applications. Front. Sustain. Food Syst.,2019, 3, DOI: 10.3389/fsufs.2019.00007 . Franco R.A., Padalhin A.R., Patrick Cuenca J., Ventura R., Montecillo A., Fernando L. & Lee B.-T., Characterization of bacterial nanocellulose produced by isolates from Philippine nata starter and its biocompatibility. J Biomater Appl., 2019, 34(3), 339–350, DOI: 10.1177/0885328219852728 . Reiniati I., Hrymak A.N. & Margaritis A., Recent developments in the production and applications of bacterial cellulose fibers and nanocrystals. Critical Reviews in Biotechnology, 2017, 37(4), 510–524, DOI: 10.1080/07388551.2016.1189871 . Huang Y., Zhu C., Yang J., Nie Y., Chen C. & Sun D., Recent advances in bacterial cellulose. Cellulose 21, 2014, DOI: 10.1007/s10570-013-0088-z . Shah N., Ul-Islam M., Khattak W.A. & Park J.K., Overview of bacterial cellulose composites: a multipurpose advanced material. Carbohydr Polym., 2013, 98(2), 1585–1598, DOI: 10.1016/j.carbpol . Nath A., Dixit M., Bandiya A., Chavda S. & Desai A.J., Enhanced PHB production and scale up studies using cheese whey in fed batch culture of Methylobacterium sp. ZP24. Bioresour. Technol., 2008, 99(13), 5749–5755. Swathi A., Sridevi V. & Rao G., Optimized lactic acid production from whey using hybrid design and ridge analysis 7.,2015. Schillinger U., Isolation and identification of lactobacilli from novel-type probiotic and mild yoghurts and their stability during refrigerated storage. Int. J. Food Microbiol., 1999, 47(1–2), 79–87, DOI: 10.1016/s0168-1605(99)00014-8 . Aasen I.M., Møretrø T., Katla T., Axelsson L. & Storrø I., Influence of complex nutrients, temperature and pH on bacteriocin production by Lactobacillus sakei CCUG 42687. Appl Microbiol Biotechnol., 2000, 53(2), 159–166, DOI: 10.1007/s002530050003 . Barajas F.H., Torres M., Arteaga L. & Castro C., GAMLSS models applied in the treatment of agro-industrial waste. RevComEst., 2015, 8(2), 245, DOI: 10.15332/s2027-3355.2015.0002.07 . Huang L.P., Jin B., Lant P. & Zhou J., Biotechnological production of lactic acid integrated with potato wastewater treatment by Rhizopus arrhizus. Journal of Chemical Technology & Biotechnology, 2003, 78(8), 899–906, DOI: 10.1002/jctb.877 . Schrecker S.T. & Gostomski P.A., Determining the Water Holding Capacity of Microbial Cellulose. Biotechnol Lett., 2005, 27(19), 1435–1438, DOI: 10.1007/s10529-005-1465-y . Kebapci K, Fragrant microcapsules, M.Sc. Thesis, Institute of Natural and Applied Sciences. Suleyman Demirel University,Isparta,2012. Cakmakci ML, Karahan AG, Cakır İ, Gündogdu A, Isolation, Molecular Diagnosis of Microorganisms to be Used in Cellulose Production and Investigation of Usage Possibilities in Microbial Cellulose Food Industry October 2008; Projet Number: 105O156. Salea R., Widjojokusumo E., Hartanti A.W., Veriansyah B. & Tjandrawinata R.R., Supercritical fluid carbon dioxide extraction of Nigella sativa (black cumin) seeds using taguchi method and full factorial design. Biochemical Compounds, 2013, 1(1), 1, DOI:2052-9341-1-1. Ranjid R.K., A primer on the Taguchi method, New York: Van Nostrand Reinhold,1990. Rutto H.L. & Enweremadu C.C., Optimization of Production Variables of Biodiesel from Manketti Using Response Surface Methodology. International Journal of Green Energy,2011, 8(7), 768–779, DOI: 10.1080/15435075.2011.600375 . Łojewska J., Miśkowiec P., Łojewski T. & Proniewicz L.M., Cellulose oxidative and hydrolytic degradation: In situ FTIR approach. Polymer Degradation and Stability, 2005, 88(3), 512–520, DOI: 10.1016/j.polymdegradstab . Cai Z. & Kim J., Preparation and Characterization of Novel Bacterial Cellulose/Gelatin Scaffold for Tissue Regeneration Using Bacterial Cellulose Hydrogel. J. Nanotechnol. Eng. Med.,2010, 1(2), DOI: 10.1115/1.4000858 . Ali M., Emsley A.M., Herman H. & Heywood R.J., Spectroscopic studies of the ageing of cellulosic paper. Polymer, 2001, 42(7), 2893–2900, DOI: 10.1016/S0032-3861(00)00691-1 . Carrillo F., Colom X., Suñol J.J. & Saurina J., Structural FTIR analysis and thermal characterisation of lyocell and viscose-type fibres. European Polymer Journal, 2004, 40(9), 2229–2234, DOI: 10.1016/j.eurpolymj.2004.05.003 . Schwanninger M., Rodrigues J.C., Pereira H. & Hinterstoisser B., Effects of short-time vibratory ball milling on the shape of FT-IR spectra of wood and cellulose. Vibrational Spectroscopy, 2004, 36(1), 23–40, DOI: 10.1016/j.vibspec.2004.02.003 . Adel A.M., Abd El-Wahab Z.H., Ibrahim A.A. & Al-Shemy M.T., Characterization of microcrystalline cellulose prepared from lignocellulosic materials. Part II: Physicochemical properties. Carbohydrate Polymers, 2010, 83(2), 676–687, DOI: 10.1016/j.carbpol.2010.08.039 . Wang L., Han G. & Zhang Y., Comparative study of composition, structure and properties of Apocynum venetum fibers under different pretreatments. Carbohydrate Polymers, 2006, 69(2), 391–397, DOI: 10.1016/j.carbpol.2006.12.028 . Klemm D., Heublein B., Fink H.-P. & Bohn A., Cellulose: fascinating biopolymer and sustainable raw material. Angew. Chem. Int. Ed. Engl., 2005, 44(22), 3358–3393, DOI: 10.1002/anie.200460587 Mangut V., Sabio E., Gañán J., González J.F., Ramiro A., González C.M., Román S. & Al-Kassir A., Thermogravimetric study of the pyrolysis of biomass residues from tomato processing industry. Fuel Processing Technology, International Congress on Energy and Environment Engineering and Management, 2006, 87(2), 109–115, DOI: 10.1016/j.fuproc.2005.08.006 . Kim U.-J., Eom S.H. & Wada M., Thermal decomposition of native cellulose: Influence on crystallite size. Polymer Degradation and Stability, 2010, 95(5), 778–781, DOI: 10.1016/j.polymdegradstab.2010.02.009 . Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-3828016\",\"acceptedTermsAndConditions\":true,\"allowDirectSubmit\":true,\"archivedVersions\":[],\"articleType\":\"Research Article\",\"associatedPublications\":[],\"authors\":[{\"id\":265053390,\"identity\":\"f5113659-5b22-4edc-91f6-7555eca2aad3\",\"order_by\":0,\"name\":\"Aytül Bayraktar\",\"email\":\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABEUlEQVRIiWNgGAWjYLCChw0gkg1E2DAwHABSPHjVMzMwJEK0MAKpNLgWCWK1HCashX9G/jGJxB12idvbj6U/+LnnvD3fjQTGB2/bGOrMG7BrkbiRzCaReCY5cc6ZtIONPc9uJ868kcBsOLeNQULmAA5rwFramBNnMKQ3NvAcuJ1gcCOBTZoXqAWXy+QhWuoTZ/A/b2z8c+CcPVAL+298WgwgWg4nzpBIO9jMc+AA4wagLcz4tBieeWxskXjmuPEMiWeJs2UOJCfOPPOwWXLOOQnJGTi0yB1PfHjj445q2Rn8aQYf3xyws+c7nnzww5syG36coSyQAKYcGxBCoPjBF5P8B8CUPW4Vo2AUjIJRMOIBAF8OYb1wjYi3AAAAAElFTkSuQmCC\",\"orcid\":\"\",\"institution\":\"Suleyman Demirel University\",\"correspondingAuthor\":true,\"prefix\":\"\",\"firstName\":\"Aytül\",\"middleName\":\"\",\"lastName\":\"Bayraktar\",\"suffix\":\"\"},{\"id\":265053391,\"identity\":\"84653c95-e16f-45b4-8377-3405ef526fcc\",\"order_by\":1,\"name\":\"Cansu Gürsoy\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Suleyman Demirel University\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Cansu\",\"middleName\":\"\",\"lastName\":\"Gürsoy\",\"suffix\":\"\"}],\"badges\":[],\"createdAt\":\"2024-01-01 19:44:38\",\"currentVersionCode\":1,\"declarations\":\"\",\"doi\":\"10.21203/rs.3.rs-3828016/v1\",\"doiUrl\":\"https://doi.org/10.21203/rs.3.rs-3828016/v1\",\"draftVersion\":[],\"editorialEvents\":[],\"editorialNote\":\"\",\"failedWorkflow\":false,\"files\":[{\"id\":49201033,\"identity\":\"d7fc047f-32fd-4a19-af7c-be591c96bc6c\",\"added_by\":\"auto\",\"created_at\":\"2024-01-05 05:04:41\",\"extension\":\"png\",\"order_by\":1,\"title\":\"Figure 1\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":77411,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eMain effect plots for S/N ratio between BNC production in \\u003cem\\u003eLactobacillus rhamnasus\\u003c/em\\u003ewith different variables. — Parcelles à effet principal pour le rapport S/N entre la production de BNC dans Lactobacillus rhamnasus avec différentes variables.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"fig1.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-3828016/v1/389de83b2b45ab3f48c277ae.png\"},{\"id\":49201317,\"identity\":\"a3fec2c1-b376-4934-a403-1f52ca8df2c2\",\"added_by\":\"auto\",\"created_at\":\"2024-01-05 05:12:41\",\"extension\":\"png\",\"order_by\":2,\"title\":\"Figure 2\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":138955,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eEffects of the parameter on the BNC production. — Effets du paramètre sur la production de BNC\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"fig2.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-3828016/v1/74320c71fba0e68572832d71.png\"},{\"id\":49201034,\"identity\":\"e765c36a-8db4-42d9-a5bb-52fdcbeec1be\",\"added_by\":\"auto\",\"created_at\":\"2024-01-05 05:04:41\",\"extension\":\"png\",\"order_by\":3,\"title\":\"Figure 3\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":46344,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eCellulose FTIR diagram produced by \\u003cem\\u003eL. rhamnosus\\u003c/em\\u003e bacteria. —Diagramme FTIR de cellulose produit par la bactérie \\u003cem\\u003eL. rhamnosus\\u003c/em\\u003e\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"fig3.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-3828016/v1/0ebb4e0fc82249eb2a4b96aa.png\"},{\"id\":49201037,\"identity\":\"48877885-1200-42f7-bc21-99c3bc609750\",\"added_by\":\"auto\",\"created_at\":\"2024-01-05 05:04:41\",\"extension\":\"png\",\"order_by\":4,\"title\":\"Figure 4\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":59118,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eViscosity of \\u003cem\\u003eL. rhamnosus \\u003c/em\\u003eBNC solution. — Viscosité de la solution \\u003cem\\u003eL. rhamnosus\\u003c/em\\u003eBNC.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"fig4.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-3828016/v1/62467cbed14156f92429d61b.png\"},{\"id\":49201038,\"identity\":\"c15b2f03-d5b5-4ae8-ba85-fc4a7bc485b1\",\"added_by\":\"auto\",\"created_at\":\"2024-01-05 05:04:41\",\"extension\":\"png\",\"order_by\":5,\"title\":\"Figure 5\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":72232,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eCellulose TGA and DTG chart obtained from \\u003cem\\u003eL. Rhamnosus.\\u003c/em\\u003e —\\u003cem\\u003e \\u003c/em\\u003eGraphique cellulose TGA et DTG obtenu auprès de\\u003cem\\u003e L. Rhamnosus\\u003c/em\\u003e\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"fig5.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-3828016/v1/33fa5d211b1c5fdd3242d40e.png\"},{\"id\":49201035,\"identity\":\"b68a4c3b-5e6f-43d3-96b0-c52dabb67e14\",\"added_by\":\"auto\",\"created_at\":\"2024-01-05 05:04:41\",\"extension\":\"png\",\"order_by\":6,\"title\":\"Figure 6\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":107841,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eFE-SEM electron microscopy images of \\u003cem\\u003eL. rhamnosus\\u003c/em\\u003e bacteria. — Images de microscopie électronique FE-SEM de la bactérie L. rhamnosus\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"fig6.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-3828016/v1/3323fa06cb4d5322551e9424.png\"},{\"id\":49934999,\"identity\":\"53b7bb86-8d74-4021-a9a6-b371cc87f851\",\"added_by\":\"auto\",\"created_at\":\"2024-01-21 18:07:40\",\"extension\":\"pdf\",\"order_by\":0,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"manuscript-pdf\",\"size\":810217,\"visible\":true,\"origin\":\"\",\"legend\":\"\",\"description\":\"\",\"filename\":\"manuscript.pdf\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-3828016/v1/aea98f65-a83a-4bba-9369-d09d1541b45e.pdf\"}],\"financialInterests\":\"No competing interests reported.\",\"formattedTitle\":\"Production of New Nano-Bacterial Cellulose with Lactobacillus rhamnosus by Using Whey Waste as Substrate with Optimization Taguchi Method, which has the potential to be used in many biomedical products\",\"fulltext\":[{\"header\":\"1. Introduction\",\"content\":\"\\u003cp\\u003eBacterial cellulose (BNC) known as a nanomaterial and could be produced by some bacteria such as Gluconacetobacter, Acetobacter spp., Rhizobium spp., Agrobacterium spp., Alcaligenes spp., Achromobacter and Sarcina [\\u003cspan citationid=\\\"CR1\\\" class=\\\"CitationRef\\\"\\u003e1\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR2\\\" class=\\\"CitationRef\\\"\\u003e2\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR3\\\" class=\\\"CitationRef\\\"\\u003e3\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR4\\\" class=\\\"CitationRef\\\"\\u003e4\\u003c/span\\u003e]. The BNC biosynthesis is a regulated process including enzymes, catalytic complexes reactions and can be investigated wih two steps such as intracellular producted of 1,4-b-glucan chains and crystallization of cellulose chains [\\u003cspan citationid=\\\"CR3\\\" class=\\\"CitationRef\\\"\\u003e3\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003eBNC is native cellulose which has high crystallinity, water capacity and strength also biocompatible with plant cellulose [\\u003cspan citationid=\\\"CR4\\\" class=\\\"CitationRef\\\"\\u003e4\\u003c/span\\u003e]. Also it produces much thinner 3D dimensional nanoporous networks with approximately 30\\u0026ndash;50 nm than plant cellulose [\\u003cspan citationid=\\\"CR5\\\" class=\\\"CitationRef\\\"\\u003e5\\u003c/span\\u003e]. It has the same molecular formula as BNC and plant celluloses (C6H10O5)n but BNC doesn\\u0026rsquo;t contain lignin, pectin, and hemicelluloses; thus BNC purification doesn\\u0026rsquo;t require harsh chemicals and also purification is easy to apply [\\u003cspan citationid=\\\"CR6\\\" class=\\\"CitationRef\\\"\\u003e6\\u003c/span\\u003e]. Having high surface area and high porosity, BNC is a material that shows high performance with other compounds in physical interaction [\\u003cspan citationid=\\\"CR3\\\" class=\\\"CitationRef\\\"\\u003e3\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR7\\\" class=\\\"CitationRef\\\"\\u003e7\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003eThis article presents an alternative strategy for the future using whey, which is waste, as well as the role of raw material for low-cost production of bacterial cellulose. Whey obtained from whey as a by-product of the dairy industry is considered a pollutant due to its high biological oxygen demand and high cost of evaluation [\\u003cspan citationid=\\\"CR8\\\" class=\\\"CitationRef\\\"\\u003e8\\u003c/span\\u003e]. Besides, it has rich nutrient content [\\u003cspan citationid=\\\"CR9\\\" class=\\\"CitationRef\\\"\\u003e9\\u003c/span\\u003e]. Lactic acid bacteria are generally used in dairy products, as pure culture. LABs are organisms that can be isolated from natural environments such as most people, animals and plants, adapted to a specific specific environment and are found in almost natural habitats. \\u003cem\\u003eLactobacillus rhamnosus\\u003c/em\\u003e strains with probiotic properties have begun to be used in new types of yoghurt [\\u003cspan citationid=\\\"CR10\\\" class=\\\"CitationRef\\\"\\u003e10\\u003c/span\\u003e]. With the increasing demand of consumers for probiotic products, the production of lactic acid bacteria in the industrial sector has gained importance [\\u003cspan citationid=\\\"CR11\\\" class=\\\"CitationRef\\\"\\u003e11\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003eIn addition to the effects of other factors in obtaining bacterial cellulose, it is crucial to determine the optimum values of some factors such as fermentation time and pH that maximize the bacterial cellulose yield [\\u003cspan citationid=\\\"CR12\\\" class=\\\"CitationRef\\\"\\u003e12\\u003c/span\\u003e]. In the present study, bacterial nano-cellulose (BNC) production was optimized based from different concentration substrate, as well as temperature, pH, and incubation time. According to the objectives of this study were determined as identifying whether the bacteria is producing \\u003cem\\u003eLactobacillus rhamnosus\\u003c/em\\u003e BNC for the first time in literature, identify an optimal media formulation for improved BNC production and characterization BNC production using fourier transform infrared spectroscopy (FT-IR) and field emission-scanning electron microscopy (FE-SEM). The substrate and process variables needed are optimized to ensure maximum BNC production at low costs. In optimization studies, while all other parameters are kept constant, one parameter is varied. In this case, it is not possible to evaluate the interaction of the parameters with each other. For this reason, Taguchi's method was used in this study to determine the effects of the factors affecting the BNC production process. Also, the evaluation of lactose in whey, which is considered as waste water with a complex organic matter content, it provides the advantage of environmentally friendly application to this study.\\u003c/p\\u003e\"},{\"header\":\"2. Materials and Methods\",\"content\":\"\\u003cdiv id=\\\"Sec3\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.1. Procurement of whey\\u003c/h2\\u003e \\u003cp\\u003eCheese whey was used as organic source and kindly obtained from Isparta \\u0026Uuml;ns\\u0026uuml;t Company (Turkey). Under aseptic conditions, samples were collected and stored at 4\\u0026deg;C in the laboratory. Then samples were stored at -20\\u0026deg;C before the analysis. It contains 60\\u0026ndash;62% lactose. The pH of the whey was adjusted to 6. Protein precipitation was induced by autoclaving from the whey at 121 oC for 20 min and after removed by filtration.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec4\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.2. Microorganism and Culture Media\\u003c/h2\\u003e \\u003cp\\u003eLyophilized culture of \\u003cem\\u003eLactobacillus rhamnasus\\u003c/em\\u003e (DSM, DELVO-ADD 100R, Netherlands) was used in all experiments. The stock culture of bacterial strain \\u003cem\\u003eLactobacillus rhamnasus\\u003c/em\\u003e was stored at \\u0026minus;\\u0026thinsp;80\\u0026deg;C in MRS medium as 15% glycerol. Precultures were per liter prepared in the 100 mL MRS culture medium containing, 1g peptone, 2g glucose, 1g beef extract, 0.2g ammonium citrate, 0.01 g magnesium sulphate, 0.5g sodium acetate, 0.005 g manganese sulphate, and 0.2 g di-potassium phosphate. A 5 mL of this culture was used for further inoculation in the 100 mL MRS production media. The media contains molasses, yeast extract, MnSO4.4H2O, Tween 80, urea, and CaCO3. Table\\u0026nbsp;\\u003cspan refid=\\\"Tab1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003e shows the design of the experiments on the set of orthogonal arrays with respect to the selected factors and levels designed with using the taguchi method. The effect of organic sources concentration, active culture, pH and incubation time on the production of bacterial cellulose was investigated using the bacteria \\u003cem\\u003eLactobacillus rhamnasus.\\u003c/em\\u003e\\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\\u003eDesign of experimental factors and levels\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/caption\\u003e \\u003ccolgroup cols=\\\"4\\\"\\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 \\u003cthead\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eFactors\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eLevel 1\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eLevel 2\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eLevel 3\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003c/thead\\u003e \\u003ctbody\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003epH\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e5.0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e5.5\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e6.0\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eOrganic source concentration\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e50% Whey\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e25% Whey\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e75% Whey\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eAmount of active culture\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e10%\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e20%\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e30%\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eIncubation time (day)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e8\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e10\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e12\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003c/tbody\\u003e \\u003c/colgroup\\u003e \\u003c/table\\u003e\\u003c/div\\u003e \\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec5\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.3. Production of BNC\\u003c/h2\\u003e \\u003cp\\u003eFermentations were performed as batch fermentations with \\u003cem\\u003eLactobacillus rhamnasus\\u003c/em\\u003e. It was cultivated in 100 mL liquid medium in 250 mL erlenmeyer flasks. Neutralizing agents were added into fermentation medium to control the pH during the fermentation. Calcium carbonate was used as neutralizing agent due to using widely for flask investigations and bioreactor processes [\\u003cspan citationid=\\\"CR13\\\" class=\\\"CitationRef\\\"\\u003e13\\u003c/span\\u003e]. Also, in this study, CaCO3 (60% (w/v) of the initial lactose concentration) was used as a neutralizing agent. The medium was inoculated with about (10% w/v) inoculum of bacteria. Fermentations were carried out with a temperature controlled orbital shaker at a stirring speed of 150 rpm and fermentation temperatures were chosen as the optimum growth temperatures given in Table\\u0026nbsp;\\u003cspan refid=\\\"Tab1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003e. All experiments were carried out in duplicate.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec6\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.4. Purification of BNC pellicles\\u003c/h2\\u003e \\u003cp\\u003eAfter BNC was produced at 30 oC for 8 days in optimum conditions, the BNC pellicles were harvested by discarding the medium and washing with large amounts of nanopure water. The pellicles were then rinsed three times with 0.1M NaOH (Sigma Aldrich, Germany) at 60\\u0026deg;C for 4 hours to remove sticky bacteria. Finally, the samples were repeatedly washed until the pH was below 7.0 with copious amounts of distilled water. Until a constant weight was reached the wet cellulose pellets were dried at 60\\u0026deg;C, then the concentration was determined as g / L (mass (g) BNC / volume of culture medium (L). For further instrumental analysis isolated pellicles were used.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec7\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.5. Characterizasyon of BNC\\u003c/h2\\u003e \\u003cp\\u003eThe The microbial cellulose obtained as a result of incubation was ground in a laboratory mill (Retsch GE) after washing and drying, and particle size was obtained as 40 mesh. Viscometer measurements were performed in cuvettes with 35 ml chambers. 3.5 g of cellulose was added and the volume of the solution was completed to 35 ml with distilled water. The viscosimetric values of this 10% suspension (Vibro Viscometer SU-10) were measured in mPa.s by taking a reading every 20 seconds in the 20\\u0026ndash;21\\u0026deg;C range, 30 Hertz frequency, constant vibration.\\u003c/p\\u003e \\u003cp\\u003eThe shaking method was used in order to measure the water holding capacity of the samples [\\u003cspan citationid=\\\"CR14\\\" class=\\\"CitationRef\\\"\\u003e14\\u003c/span\\u003e]. Two shakes were made and weighed to remove excess water from the surface of the samples. The samples were dried at the room temperature for 48 hours and then dried at 60\\u0026deg;C for 12 hours to completely remove the water content. The water holding capacity was calculated using the following method:\\u003cdiv id=\\\"Equ1\\\" class=\\\"Equation\\\"\\u003e\\u003cdiv format=\\\"TEX\\\" class=\\\"mathdisplay\\\" id=\\\"FileID_Equ1\\\" name=\\\"EquationSource\\\"\\u003e\\n$$\\\\text{W}\\\\text{a}\\\\text{t}\\\\text{e}\\\\text{r} \\\\text{H}\\\\text{o}\\\\text{l}\\\\text{d}\\\\text{i}\\\\text{n}\\\\text{g} \\\\text{C}\\\\text{a}\\\\text{p}\\\\text{a}\\\\text{c}\\\\text{i}\\\\text{t}\\\\text{y}=\\\\frac{\\\\text{M}\\\\text{a}\\\\text{s}\\\\text{s} \\\\text{o}\\\\text{f} \\\\text{w}\\\\text{a}\\\\text{t}\\\\text{e}\\\\text{r} \\\\text{r}\\\\text{e}\\\\text{m}\\\\text{o}\\\\text{v}\\\\text{e}\\\\text{d} \\\\text{d}\\\\text{u}\\\\text{r}\\\\text{i}\\\\text{n}\\\\text{g} \\\\text{d}\\\\text{r}\\\\text{y}\\\\text{i}\\\\text{n}\\\\text{g} \\\\left(\\\\text{g}\\\\right)}{\\\\text{D}\\\\text{r}\\\\text{y} \\\\text{w}\\\\text{e}\\\\text{i}\\\\text{g}\\\\text{h}\\\\text{t} \\\\text{o}\\\\text{f} \\\\text{b}\\\\text{a}\\\\text{c}\\\\text{t}\\\\text{e}\\\\text{r}\\\\text{i}\\\\text{a}\\\\text{l} \\\\text{n}\\\\text{a}\\\\text{n}\\\\text{o}-\\\\text{c}\\\\text{e}\\\\text{l}\\\\text{l}\\\\text{u}\\\\text{l}\\\\text{o}\\\\text{s}\\\\text{e} \\\\left(\\\\text{g}\\\\right)}$$\\u003c/div\\u003e\\u003cdiv class=\\\"EquationNumber\\\"\\u003e1\\u003c/div\\u003e\\u003c/div\\u003e\\u003c/p\\u003e \\u003cp\\u003eThermal gravimetric analysis was performed to measure the weight loss and BNC thermal stability of the analyzed material when the temperature increased linearly [\\u003cspan citationid=\\\"CR15\\\" class=\\\"CitationRef\\\"\\u003e15\\u003c/span\\u003e]. Thermal degradation behavior of bacterial cellulose was investigated at 0-900 oC using the TGA / TDA device (Perkin Elmer diamond, USA). Percent and derivative weight loss were recorded against temperature for all samples.\\u003c/p\\u003e \\u003cp\\u003eSolubility characteristics of microbial cellulose were determined using various solvents. The solvents used for solubility tests are toluene, acetone, chloroform, benzene, carbon tetrachloride, and dimethylformamide. In the experiment, 0.1 g of sample was placed in 10 mL of solvent and the tubes were rinsed at room temperature for 24 h to determine their solubility in solvents [\\u003cspan citationid=\\\"CR16\\\" class=\\\"CitationRef\\\"\\u003e16\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003eTo determine the elemental contents of the BNC samples obtained as a result of experimental studies, FT-IR Spectroscopy was evaluated. It has been concluded whether pure cellulose is obtained from the numerical percentage of the elemental C content in the FT-IR bands of the cellulose sample. FT-IR analysis was carried out in S\\u0026uuml;leyman Demirel University Experimental and Observational Student Research and Application Center Laboratory. FT-IR spectra of the BNC pellicle were obtained with Perkin-Elmer FT-IR spectrophotometer (Norwalk, USA) in diffuse reflectance mode at a resolution of 4 particles/cm in KBr pellets.\\u003c/p\\u003e \\u003cp\\u003eFe-SEM was used to determine the surface structure of BNC cellulose samples obtained after drying for 10 days at 28\\u0026deg;C in the liquid medium. Fe-SEM analysis was carried out at Isparta S\\u0026uuml;leyman Demirel University Energy Research and Research Application Center. Microstructures and morphology of samples were evaluated by examining images in three dimensions with TEM..\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec8\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.6. Taguchi Methodology\\u003c/h2\\u003e \\u003cp\\u003eTaguchi is one of the impressive ways to design an experiment to optimize the process. Unlike traditional full factor research, it can be used as a powerful tool to optimize performance characteristics of process parameters. The Taguchi method uses orthogonal array design (OAD) while designing less number of experiments with a large number of parameters [\\u003cspan citationid=\\\"CR17\\\" class=\\\"CitationRef\\\"\\u003e17\\u003c/span\\u003e]. The signal / noise (S / N) ratio enables the quality characteristics of the process to be measured precisely at different process levels of controllable and uncontrollable factors. In addition, it makes it possible to minimize the negative effects of uncontrollable factors on the process. It is possible to provide the optimum parameters of each parameter in the process by determining the S / N ratios.\\u003c/p\\u003e \\u003cp\\u003eThe measurement of the quality of the results is measured by the signal-to-noise (S / N) ratio with three different characteristics of the target values: \\u0026ldquo;larger is better\\u0026rdquo;, \\u0026ldquo;smaller is better\\u0026rdquo; or \\u0026ldquo;nominal is better\\u0026rdquo; [\\u003cspan citationid=\\\"CR18\\\" class=\\\"CitationRef\\\"\\u003e18\\u003c/span\\u003e]. In this study, target values of \\u0026ldquo;smaller is better\\u0026rdquo; was chosen. Because it is aimed to determine the highest production at the lowest concentrations of the parameters. Thus, it is determined how the effects of the selected parameters at the lowest levels affect the BNC production efficiency. In this study, interactions between factors were not taken into account and the main effects of four parameters affecting the process were evaluated.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec9\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.7. Design of Experiments and Choice of Factors\\u003c/h2\\u003e \\u003cp\\u003eIn this study the Taguchi method was used as an experimental design and analysis method. The L9 orthogonal arrays improved by Taguchi method. The parameters determined by examining the literature information and their levels are given in Table\\u0026nbsp;\\u003cspan refid=\\\"Tab2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003e. BNC production was determined by optimization conditions, four different variables were selected as pH, carbon sources concentrations, amount of active culture conditions, and incubation time. Also,3 different levels were determined for each parameter. The main steps of this method was created firstly determination of factors, interactions and the levels of each factor. Minitab 16 software (Minitab Inc., USA) was used to create the experimental design and evaluate the data. To determine the optimization of the BNC manufacturing process, how various factors influence the process individually as well as how they contribute to the complexity of the entire process and their multiple interactions were also evaluated. By measuring the predicted interactions of different factors (based on the severity index (SI)), the effects of four different factors used in the experiment set at various levels were determined. Data obtained from Taguchi design experiment and all parameters were subjected to analysis of variance (ANOVA) for the significant difference between BNC production efficiency. P values less than 0.05 were considered significant. All statistical analyzes were performed using MINITAB v.16 (Minitab Inc., USA) statistical software package.\\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\\u003eTaguchi L9 orthogonal experimental design\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/caption\\u003e \\u003ccolgroup cols=\\\"5\\\"\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c1\\\" colnum=\\\"1\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"char\\\" char=\\\".\\\" class=\\\"colspec\\\" colname=\\\"c2\\\" colnum=\\\"2\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"char\\\" char=\\\".\\\" class=\\\"colspec\\\" colname=\\\"c3\\\" colnum=\\\"3\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"char\\\" char=\\\".\\\" class=\\\"colspec\\\" colname=\\\"c4\\\" colnum=\\\"4\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"char\\\" char=\\\".\\\" class=\\\"colspec\\\" colname=\\\"c5\\\" colnum=\\\"5\\\"\\u003e\\u003c/div\\u003e \\u003cthead\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eExperiment No\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003epH\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eOrganic source concentration\\u003c/p\\u003e \\u003cp\\u003e(%)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eActive culture\\u003c/p\\u003e \\u003cp\\u003e(%)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eIncubation time\\u003c/p\\u003e \\u003cp\\u003e(day)\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003c/thead\\u003e \\u003ctbody\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e5.0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e10\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e8\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e2\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e5.0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e50\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e20\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e10\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e3\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e5.0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e75\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e30\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e12\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e4\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e5.5\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e20\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e12\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e5\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e5.5\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e50\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e30\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e8\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e6\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e5.5\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e75\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e10\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e10\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e7\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e6.0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e30\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e10\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e8\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e6.0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e50\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e10\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e12\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e9\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e6.0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e75\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e20\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e8\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003c/tbody\\u003e \\u003c/colgroup\\u003e \\u003c/table\\u003e\\u003c/div\\u003e \\u003c/p\\u003e \\u003c/div\\u003e\"},{\"header\":\"3. Results and discussion\",\"content\":\"\\u003cdiv id=\\\"Sec11\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e3.1. Taguchi Method\\u003c/h2\\u003e \\u003cp\\u003eIn In this study, three separate levels were defined for each of the BNC production process parameters and 81 separate experiments were required to complete the process. Instead, experiments were carried out by applying the L9 orthogonal array scheme, which requires nine experiments with the taguchi method. Experimental results were evaluated according to the Taguchi OA L9 method. Figure\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003e. shows main effect plots for S/N ratio for BNC production in \\u003cem\\u003eLactobacillus rhamnasus\\u003c/em\\u003e with different variables, including pH (5\\u0026ndash;6), organic source concentration (25\\u0026ndash;100), active culture (10\\u0026ndash;30), incubation period (8\\u0026ndash;12). The main effect graphs showing the signal-to-noise (S / N) ratio describing the scattering around a target value were obtained by using Taguchi method. Noise factors are seen as the reason for the variability of the answers examined for the target value. S / N ratio measures the performance level and the effects of noise factors on performance. The variance decomposition is obtained for the relative effects of different production parameters on BNC productivity yield, which is called variance analysis (ANOVA) [\\u003cspan citationid=\\\"CR19\\\" class=\\\"CitationRef\\\"\\u003e19\\u003c/span\\u003e]. The ANOVA was investigated to find the effects of process parameters on BNC production. Different levels of BNC production were evaluated the effect of individual parameters for the conversion. According to Taguchi Method, the optimum \\u003cem\\u003eLactobacillus rhamnasus\\u003c/em\\u003e production conditions was obtained at pH 5.5, organic source concentration 25%, active culture 30%, incubation period 8 days.\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003cp\\u003eIn the case of the design with the Taguchi method, when the experimental data and the estimation data were examined, 4.3 g BNC yield was obtained at pH 5.8 and the organic source concentration was 40%. While, the active culture amount was 10% at pH 5.5, 5 g BNC yield was obtained. If the organic source concentration was 70% and the active culture amount was 25%, the amount of BNC yield was 2 g. When the organic source concentration was 70% and the incubation time selected as 10 days, the amount of BNC yield was 2 g. 0.2 g BNC yield was obtained with 20%amount of active culture and 10 days incubation time.\\u003c/p\\u003e \\u003cp\\u003eThe Taguchi method based on OA detects optimum levels of process parameters by reducing variance for experiments. The results were then converted into S / N ratio data. S / N ratio of Taguchi method was used for data analysis and estimation of optimum parameters. Taguchi method factor design gives good results in the effect studies of various combinations. Figure\\u0026nbsp;\\u003cspan refid=\\\"Fig2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003e. shows the effects of four parameters such as pH, organic source concentration, incubation time and active culture concentration on BNC yield using Taguchi experimental design. pH and incubation times have been shown to positive effect on production efficiency by affecting BNC production. The effect of organic source concentration varies depending on the amount of active culture. Increasing in the amount of active culture and increasing the number of individuals in the medium caused a decrease in BNC efficiency due to the insufficient organic source in the medium. The relative interactions of the factors on the BNC process performance are shown in Table\\u0026nbsp;\\u003cspan refid=\\\"Tab3\\\" class=\\\"InternalRef\\\"\\u003e3\\u003c/span\\u003e. Table\\u0026nbsp;\\u003cspan refid=\\\"Tab3\\\" class=\\\"InternalRef\\\"\\u003e3\\u003c/span\\u003e shows that tested model has shown that model effectiveness was found as R2\\u0026thinsp;=\\u0026thinsp;96.22%. Analysis of variance (ANOVA) determined the significance of each independent variable which was presented in (Table\\u0026nbsp;\\u003cspan refid=\\\"Tab4\\\" class=\\\"InternalRef\\\"\\u003e4\\u003c/span\\u003e). With 95% confidence interval, all parameters have marginally significant effects (P\\u0026thinsp;\\u0026gt;\\u0026thinsp;0.05).\\u003c/p\\u003e \\u003cp\\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\\u003eGeneral Linear Model: BNC value versus pH; organic source, active culture analysis of variance for BNC value, using sequential SS for tests.\\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\\u003eSource\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eDF\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eSeq SS\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eAdj SS\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eSeq MS\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003eF\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003eP\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003c/thead\\u003e \\u003ctbody\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003epH\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e2\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e4.6769\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e4.6769\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e2.3384\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e2.49\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003e0.409\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eOrganic source concentration\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e2\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e9.1785\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e9.1785\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e4.5892\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e4.89\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003e0.305\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eActive culture\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e2\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e4.4193\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e4.4193\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e2.2096\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e2.35\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003e0.419\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eIncubation period\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e5.5873\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e5.5873\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e5.5873\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e5.95\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003e0.248\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eError\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e0.9384\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0.9384\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e0.9384\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eTotal\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e8\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e24.8004\\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 \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"7\\\" nameend=\\\"c7\\\" namest=\\\"c1\\\"\\u003e \\u003cp\\u003eS\\u0026thinsp;=\\u0026thinsp;0.968736 R-Sq\\u0026thinsp;=\\u0026thinsp;96.22% R-Sq(adj)\\u0026thinsp;=\\u0026thinsp;69.73%\\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 \\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\\u003eAnalysis of variance (ANOVA) evaluation on BNC yield.\\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\\u003eExperiment No\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003epH\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eOrganic Source Concentration\\u003c/p\\u003e \\u003cp\\u003e(g/L%)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eActive Culture\\u003c/p\\u003e \\u003cp\\u003e(g/L%)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eIncubation time\\u003c/p\\u003e \\u003cp\\u003e(day)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003eBNC value\\u003c/p\\u003e \\u003cp\\u003e(g)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003eSNRA1\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003c/thead\\u003e \\u003ctbody\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e5.0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e10\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e8\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e0.06\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003e24.4370\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e2\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e5.0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e50\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e20\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e10\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e1.13\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003e-1.0616\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e3\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e5.0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e75\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e30\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e12\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e0.67\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003e3.4785\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e4\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e5.5\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e20\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e12\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e0.00\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003e*\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e5\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e5.5\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e50\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e30\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e8\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e0.23\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003e12.7654\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e6\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e5.5\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e75\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e10\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e10\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e0.00\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003e*\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e7\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e6.0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e30\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e10\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e0.00\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003e*\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e8\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e6.0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e50\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e10\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e12\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e5.41\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003e-14.6639\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e9\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e6.0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e75\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e20\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e8\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e0.00\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003e*\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003c/tbody\\u003e \\u003c/colgroup\\u003e \\u003c/table\\u003e\\u003c/div\\u003e \\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec12\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e3.2. FTIR\\u003c/h2\\u003e \\u003cp\\u003eFigure \\u003cspan refid=\\\"Fig3\\\" class=\\\"InternalRef\\\"\\u003e3\\u003c/span\\u003e indicates FTIR spectra of L. rhamnosus cellulose. According to the analysis results, the broad band intramolecular and intermolecular hydrogen bonding -OH stretching at 3430 cm-1 observed in the FTIR spectrum of the BNC sample obtained from L. rhamnosus.\\u003c/p\\u003e \\u003cp\\u003eOther researchers also found major stretching peak associated with \\u0026ndash;OH groups on the glucose rings and water molecules take place between 3300 cm-1 and 3600 cm-1 [\\u003cspan citationid=\\\"CR20\\\" class=\\\"CitationRef\\\"\\u003e20\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR21\\\" class=\\\"CitationRef\\\"\\u003e21\\u003c/span\\u003e]. Ali et al., (2001) found that the C-H stretching vibration bands of -CH3 and \\u0026ndash;CH2 groups at 2926 cm-1 whereas bending bands of these groups at 1458 and 1562 cm-1. Also, in our study the intensities of bands at 2926 cm-1 and 1628 cm-1 were observed for L. rhamnosus cellulose [\\u003cspan citationid=\\\"CR22\\\" class=\\\"CitationRef\\\"\\u003e22\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003cp\\u003eFTIR results showed that intensities of bands at 2926 cm-1 and 1628 cm-1 were monitored for L. rhamnosus cellulose. This peak intensity is indicative of the existence of more aliphatic carbon groups (-CH2-). These spectra of standard and produced cellulose showed that these compounds obtained from different media. The band at 1628 cm-1 is associated with water in cellulose and probably some hemicelluloses [\\u003cspan citationid=\\\"CR23\\\" class=\\\"CitationRef\\\"\\u003e23\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR24\\\" class=\\\"CitationRef\\\"\\u003e24\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR25\\\" class=\\\"CitationRef\\\"\\u003e25\\u003c/span\\u003e]. In a study with microcrystalline cellulose and bamboo fibers, the H-bound OH groups were seen at 3340\\u0026ndash;3412 cm-1 wave length, and the broader band gap CH tension was similaries to our study (2968\\u0026thinsp;\\u0026minus;\\u0026thinsp;2900 cm-1) [\\u003cspan citationid=\\\"CR26\\\" class=\\\"CitationRef\\\"\\u003e26\\u003c/span\\u003e].\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec13\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e3.3. Water Release Feature\\u003c/h2\\u003e \\u003cp\\u003eThe BNC yield was found as 60.4 g/g for L. rhamnosus bacterium. Water release rate of cellulose produced by L. rhamnosus bacteria was determined as 64.4%. The water release rate of celluloses during the drying period was determined as 28.4%. The water release process of the cellulose produced by the L. rhamnosus bacterium at the end of the 7 hours. The WHC of the BNCs produced in this study was in a suitable agreement with literature-cited publications [\\u003cspan citationid=\\\"CR27\\\" class=\\\"CitationRef\\\"\\u003e27\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003eThe viscosity value of L. rhamnosus BS solution was found as 36 mPa.s at 0. Seconds (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig4\\\" class=\\\"InternalRef\\\"\\u003e4\\u003c/span\\u003e.). A linear decrease in viscosity values was observed by dilution of BS solution. It was determined that the viscosity decreased from 36 mPa.s to 31.2 mPa.s within 3 minute.\\u003c/p\\u003e \\u003cp\\u003eThe WHC (water holding capacity) was considered as an most important physical characteristics due to biomedical application of BNC. The variations between the WHC were related with porosity and surface area. The greater surface area and larger pore size allow more water to penetrate and indwell in the BNC matrix.\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec14\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e3.4. Thermal properties\\u003c/h2\\u003e \\u003cp\\u003eFigure \\u003cspan refid=\\\"Fig5\\\" class=\\\"InternalRef\\\"\\u003e5\\u003c/span\\u003e. shows the thermogravimetric /differential thermal analysis (TG/DTG) curves of L. rhamnosus BNC sample. The thermal degradation behaviors of cellulose produced by L. rhamnosus were investigated and the temperatures of decomposition were determined. The graphic in Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig5\\\" class=\\\"InternalRef\\\"\\u003e5\\u003c/span\\u003e shows that thermal degradation occurs in two different stages.\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003cp\\u003eMangut et al. (2006) found that first degradation step was started at 250\\u0026deg;C and ended about 430\\u0026deg;C and peak was obtained at 350\\u0026deg;C. It means that it is cellulosic materials. At this temperature, simple structures such as CO, CO2, H2O, CH4, CH3OH are being removed by decarboxylation and demethoxylation. The second decay was observed at 780\\u0026deg;C [\\u003cspan citationid=\\\"CR28\\\" class=\\\"CitationRef\\\"\\u003e28\\u003c/span\\u003e]. In this step, the aromatic components are probably separated more slowly. Due to the pyrolysis and decomposition of the cellulose concentration it is seen that a slower weight loss from 350\\u0026deg;C to 700\\u0026deg;C to form a crosslinking ring structure. Many studies have shown the behavior of cellulose in inorganic and organic solvents. Although the ability to dissolve in various solvents is of importance for the usage of BNC in different industry, it is also important to reveal the effects of organism differences on the cellulosic composition. Toluene, acetone, chloroform, benzene, carbon tetrachloride and dimethylformamide solvents were used for solubility tests of the cellulose samples produced by L rhamnasus bacterium.\\u003c/p\\u003e \\u003cp\\u003eIn the experiment, 0.1 g sample was placed in 10 ml of solvent and the tubes were shaken for 24 hours at room temperature, and their dissolution ability in the solvents was determined. As a result of the solubility test, it was determined that BNC was dissolved only in toluene solvent in 80oC water bath. Solubility, breaking hydrogen bonds between polymer chains and increasing the accessibility to the intercrystalline structure can be explained.\\u003c/p\\u003e \\u003cp\\u003eThe DTG curve for L. rhamnosus BNC sample showed that higher temperatures was shifted by increasing crystallite size. This behavior suggests that higher crystallite size shows higher thermal stability. Kim et al. (2010) also found that an increase in the crystallite size promoted by higher thermal stability. Therefore, BNC sample has probably higher thermal stability because it has higher amount of hydrogen bonds between cellulose chains and leads to more ordered and packed cellulose regions [\\u003cspan citationid=\\\"CR29\\\" class=\\\"CitationRef\\\"\\u003e29\\u003c/span\\u003e]. High crystallite BNC can possibly turn into a increasing of the thermal decomposition temperature of cellulose. The intensity of L. Rhamnosus BNC, which is believed to be rich in aromatic structures, supports the FTIR analysis results.\\u003c/p\\u003e \\u003cp\\u003eFigure \\u003cspan refid=\\\"Fig6\\\" class=\\\"InternalRef\\\"\\u003e6\\u003c/span\\u003e. shows the cellulosic structure produced by FE-SEM electron microscopy images of L. rhamnosus bacteria with x400 and x24000 magnification, respectively. Figure\\u0026nbsp;\\u003cspan refid=\\\"Fig6\\\" class=\\\"InternalRef\\\"\\u003e6\\u003c/span\\u003e. shows that it was determined that the cellulosic fibers formed a tubular network structure. The cellulose fibers in these images were reinforced by determining their chemical and physical properties.\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003c/div\\u003e\"},{\"header\":\"4. Conclusion\",\"content\":\"\\u003cp\\u003eIn this study, the production of BNC was optimized using different variables such as pH, organic source concentration, active culture concentration, and incubation period by Taguchi method. Characterization tests showed that the produced BNC revealed a strong network of nano fibrils. In addition, it does not involve toxic and any kind of hazardous materials in producing BNC, which is excellent and suitable for safe environments such as medical, a packing material in fields such as wound healing, drug coating, vein production, food industry and cosmetics applications. In conclusion, BNC produced from L. rhamnosus could play an crucial and environmentally friendly role in diverse medical and industrial applications as another way to cellulose obtained from other sources, such as microorganism and plants.\\u003c/p\\u003e\"},{\"header\":\"Declarations\",\"content\":\"\\u003cp\\u003eEthical approval is not required. There is no funding received. The dataset was not used.\\u003c/p\\u003e\\n\\u003ch2\\u003eAuthor Contribution\\u003c/h2\\u003e\\n\\u003cp\\u003eA.B. and C.D. wrote the body of the manuscript and all authors reviewed the manuscript.\\u003c/p\\u003e\"},{\"header\":\"References\",\"content\":\"\\u003col\\u003e\\u003cli\\u003e\\u003cspan\\u003eIqbal H.M.N., Kyazze G., Locke I.C., Tron T. \\u0026amp; Keshavarz T., Development of bio-composites with novel characteristics: Evaluation of phenol-induced antibacterial, biocompatible and biodegradable behaviours, Carbohydrate Polymers,2015, 131, 197\\u0026ndash;207.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eGallegos A.M.A., Herrera Carrera S., Parra R., Keshavarz T. \\u0026amp; Iqbal H.M.N., Bacterial Cellulose: A Sustainable Source to Develop Value-Added Products \\u0026ndash; A Review. BioResources,2016, 11(2), 5641\\u0026ndash;5655, DOI:\\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003e10.15376/biores.11.2.Gallegos\\u003c/span\\u003e\\u003cspan address=\\\"10.15376/biores.11.2.Gallegos\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eAzeredo H.M.C., Barud H., Farinas C.S., Vasconcellos V.M. \\u0026amp; Claro A.M., Bacterial Cellulose as a Raw Material for Food and Food Packaging Applications. Front. Sustain. Food Syst.,2019, 3, DOI:\\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003e10.3389/fsufs.2019.00007\\u003c/span\\u003e\\u003cspan address=\\\"10.3389/fsufs.2019.00007\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eFranco R.A., Padalhin A.R., Patrick Cuenca J., Ventura R., Montecillo A., Fernando L. \\u0026amp; Lee B.-T., Characterization of bacterial nanocellulose produced by isolates from Philippine nata starter and its biocompatibility. J Biomater Appl., 2019, 34(3), 339\\u0026ndash;350, DOI:\\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003e10.1177/0885328219852728\\u003c/span\\u003e\\u003cspan address=\\\"10.1177/0885328219852728\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eReiniati I., Hrymak A.N. \\u0026amp; Margaritis A., Recent developments in the production and applications of bacterial cellulose fibers and nanocrystals. Critical Reviews in Biotechnology, 2017, 37(4), 510\\u0026ndash;524, DOI:\\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003e10.1080/07388551.2016.1189871\\u003c/span\\u003e\\u003cspan address=\\\"10.1080/07388551.2016.1189871\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eHuang Y., Zhu C., Yang J., Nie Y., Chen C. \\u0026amp; Sun D., Recent advances in bacterial cellulose. Cellulose 21, 2014, DOI:\\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003e10.1007/s10570-013-0088-z\\u003c/span\\u003e\\u003cspan address=\\\"10.1007/s10570-013-0088-z\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eShah N., Ul-Islam M., Khattak W.A. \\u0026amp; Park J.K., Overview of bacterial cellulose composites: a multipurpose advanced material. Carbohydr Polym., 2013, 98(2), 1585\\u0026ndash;1598, DOI:\\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003e10.1016/j.carbpol\\u003c/span\\u003e\\u003cspan address=\\\"10.1016/j.carbpol\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eNath A., Dixit M., Bandiya A., Chavda S. \\u0026amp; Desai A.J., Enhanced PHB production and scale up studies using cheese whey in fed batch culture of Methylobacterium sp. ZP24. Bioresour. Technol., 2008, 99(13), 5749\\u0026ndash;5755.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eSwathi A., Sridevi V. \\u0026amp; Rao G., Optimized lactic acid production from whey using hybrid design and ridge analysis 7.,2015.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eSchillinger U., Isolation and identification of lactobacilli from novel-type probiotic and mild yoghurts and their stability during refrigerated storage. Int. J. Food Microbiol., 1999, 47(1\\u0026ndash;2), 79\\u0026ndash;87, DOI:\\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003e10.1016/s0168-1605(99)00014-8\\u003c/span\\u003e\\u003cspan address=\\\"10.1016/s0168-1605(99)00014-8\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eAasen I.M., M\\u0026oslash;retr\\u0026oslash; T., Katla T., Axelsson L. \\u0026amp; Storr\\u0026oslash; I., Influence of complex nutrients, temperature and pH on bacteriocin production by Lactobacillus sakei CCUG 42687. Appl Microbiol Biotechnol., 2000, 53(2), 159\\u0026ndash;166, DOI:\\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003e10.1007/s002530050003\\u003c/span\\u003e\\u003cspan address=\\\"10.1007/s002530050003\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eBarajas F.H., Torres M., Arteaga L. \\u0026amp; Castro C., GAMLSS models applied in the treatment of agro-industrial waste. RevComEst., 2015, 8(2), 245, DOI:\\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003e10.15332/s2027-3355.2015.0002.07\\u003c/span\\u003e\\u003cspan address=\\\"10.15332/s2027-3355.2015.0002.07\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eHuang L.P., Jin B., Lant P. \\u0026amp; Zhou J., Biotechnological production of lactic acid integrated with potato wastewater treatment by Rhizopus arrhizus. Journal of Chemical Technology \\u0026amp; Biotechnology, 2003, 78(8), 899\\u0026ndash;906, DOI:\\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003e10.1002/jctb.877\\u003c/span\\u003e\\u003cspan address=\\\"10.1002/jctb.877\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eSchrecker S.T. \\u0026amp; Gostomski P.A., Determining the Water Holding Capacity of Microbial Cellulose. Biotechnol Lett., 2005, 27(19), 1435\\u0026ndash;1438, DOI:\\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003e10.1007/s10529-005-1465-y\\u003c/span\\u003e\\u003cspan address=\\\"10.1007/s10529-005-1465-y\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eKebapci K, Fragrant microcapsules, M.Sc. Thesis, Institute of Natural and Applied Sciences. Suleyman Demirel University,Isparta,2012.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eCakmakci ML, Karahan AG, Cakır İ, G\\u0026uuml;ndogdu A, Isolation, Molecular Diagnosis of Microorganisms to be Used in Cellulose Production and Investigation of Usage Possibilities in Microbial Cellulose Food Industry October 2008; Projet Number: 105O156.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eSalea R., Widjojokusumo E., Hartanti A.W., Veriansyah B. \\u0026amp; Tjandrawinata R.R., Supercritical fluid carbon dioxide extraction of Nigella sativa (black cumin) seeds using taguchi method and full factorial design. Biochemical Compounds, 2013, 1(1), 1, DOI:2052-9341-1-1.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eRanjid R.K., A primer on the Taguchi method, New York: Van Nostrand Reinhold,1990.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eRutto H.L. \\u0026amp; Enweremadu C.C., Optimization of Production Variables of Biodiesel from Manketti Using Response Surface Methodology. International Journal of Green Energy,2011, 8(7), 768\\u0026ndash;779, DOI:\\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003e10.1080/15435075.2011.600375\\u003c/span\\u003e\\u003cspan address=\\\"10.1080/15435075.2011.600375\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eŁojewska J., Miśkowiec P., Łojewski T. \\u0026amp; Proniewicz L.M., Cellulose oxidative and hydrolytic degradation: In situ FTIR approach. Polymer Degradation and Stability, 2005, 88(3), 512\\u0026ndash;520, DOI:\\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003e10.1016/j.polymdegradstab\\u003c/span\\u003e\\u003cspan address=\\\"10.1016/j.polymdegradstab\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eCai Z. \\u0026amp; Kim J., Preparation and Characterization of Novel Bacterial Cellulose/Gelatin Scaffold for Tissue Regeneration Using Bacterial Cellulose Hydrogel. J. Nanotechnol. Eng. Med.,2010, 1(2), DOI:\\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003e10.1115/1.4000858\\u003c/span\\u003e\\u003cspan address=\\\"10.1115/1.4000858\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eAli M., Emsley A.M., Herman H. \\u0026amp; Heywood R.J., Spectroscopic studies of the ageing of cellulosic paper. Polymer, 2001, 42(7), 2893\\u0026ndash;2900, DOI:\\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003e10.1016/S0032-3861(00)00691-1\\u003c/span\\u003e\\u003cspan address=\\\"10.1016/S0032-3861(00)00691-1\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eCarrillo F., Colom X., Su\\u0026ntilde;ol J.J. \\u0026amp; Saurina J., Structural FTIR analysis and thermal characterisation of lyocell and viscose-type fibres. European Polymer Journal, 2004, 40(9), 2229\\u0026ndash;2234, DOI:\\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003e10.1016/j.eurpolymj.2004.05.003\\u003c/span\\u003e\\u003cspan address=\\\"10.1016/j.eurpolymj.2004.05.003\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eSchwanninger M., Rodrigues J.C., Pereira H. \\u0026amp; Hinterstoisser B., Effects of short-time vibratory ball milling on the shape of FT-IR spectra of wood and cellulose. Vibrational Spectroscopy, 2004, 36(1), 23\\u0026ndash;40, DOI:\\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003e10.1016/j.vibspec.2004.02.003\\u003c/span\\u003e\\u003cspan address=\\\"10.1016/j.vibspec.2004.02.003\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eAdel A.M., Abd El-Wahab Z.H., Ibrahim A.A. \\u0026amp; Al-Shemy M.T., Characterization of microcrystalline cellulose prepared from lignocellulosic materials. Part II: Physicochemical properties. Carbohydrate Polymers, 2010, 83(2), 676\\u0026ndash;687, DOI:\\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003e10.1016/j.carbpol.2010.08.039\\u003c/span\\u003e\\u003cspan address=\\\"10.1016/j.carbpol.2010.08.039\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eWang L., Han G. \\u0026amp; Zhang Y., Comparative study of composition, structure and properties of Apocynum venetum fibers under different pretreatments. Carbohydrate Polymers, 2006, 69(2), 391\\u0026ndash;397, DOI:\\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003e10.1016/j.carbpol.2006.12.028\\u003c/span\\u003e\\u003cspan address=\\\"10.1016/j.carbpol.2006.12.028\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eKlemm D., Heublein B., Fink H.-P. \\u0026amp; Bohn A., Cellulose: fascinating biopolymer and sustainable raw material. Angew. Chem. Int. Ed. Engl., 2005, 44(22), 3358\\u0026ndash;3393, DOI:\\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003e10.1002/anie.200460587\\u003c/span\\u003e\\u003cspan address=\\\"10.1002/anie.200460587\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eMangut V., Sabio E., Ga\\u0026ntilde;\\u0026aacute;n J., Gonz\\u0026aacute;lez J.F., Ramiro A., Gonz\\u0026aacute;lez C.M., Rom\\u0026aacute;n S. \\u0026amp; Al-Kassir A., Thermogravimetric study of the pyrolysis of biomass residues from tomato processing industry. Fuel Processing Technology, International Congress on Energy and Environment Engineering and Management, 2006, 87(2), 109\\u0026ndash;115, DOI:\\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003e10.1016/j.fuproc.2005.08.006\\u003c/span\\u003e\\u003cspan address=\\\"10.1016/j.fuproc.2005.08.006\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eKim U.-J., Eom S.H. \\u0026amp; Wada M., Thermal decomposition of native cellulose: Influence on crystallite size. Polymer Degradation and Stability, 2010, 95(5), 778\\u0026ndash;781, DOI:\\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003e10.1016/j.polymdegradstab.2010.02.009\\u003c/span\\u003e\\u003cspan address=\\\"10.1016/j.polymdegradstab.2010.02.009\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e.\\u003c/span\\u003e\\u003c/li\\u003e\\u003c/ol\\u003e\"}],\"fulltextSource\":\"\",\"fullText\":\"\",\"funders\":[],\"hasAdminPriorityOnWorkflow\":false,\"hasManuscriptDocX\":true,\"hasOptedInToPreprint\":true,\"hasPassedJournalQc\":\"\",\"hasAnyPriority\":true,\"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\":\"info@researchsquare.com\",\"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\":\"Lactobacillus rhamnosus, bacterial cellulose, taguchi design, whey waste\",\"lastPublishedDoi\":\"10.21203/rs.3.rs-3828016/v1\",\"lastPublishedDoiUrl\":\"https://doi.org/10.21203/rs.3.rs-3828016/v1\",\"license\":{\"name\":\"CC BY 4.0\",\"url\":\"https://creativecommons.org/licenses/by/4.0/\"},\"manuscriptAbstract\":\"\\u003cp\\u003eWhey waste, which has a negative impact on the environment, is an important component with high organic content. The fact that it contains lactose, a fermentable sugar, is a suitable substrate for the formation of natural nano-cellulose. Bacterial nano-cellulose (BNC), a type of natural cellulose polymer synthesized by some microorganisms, has been reported to be a promising natural biomedical material due to its distinctive feature, including its unique fibril nanostructure, high water holding capacity, crystallinity, high chemical purity, fine wet mechanical property.\\u003c/p\\u003e\\n\\u003cp\\u003eIn this study, new BNC production was realized for the first time by using \\u003cem\\u003eLactobacillus rhamnosus\\u003c/em\\u003e bacteria and whey as organic substrate. Optimum condition was determined by Taguchi method under the following condition; pH (5-6), organic source concentration (25-100 % g/L), active culture (10-30 % g/L), incubation period (8-12 day). Whereas Taguchi method was highest performed at at pH 5.5, organic source concentration 25 % g/L, active culture 30 % g/L, incubation period 8 days with 5.41 g BNC yield. Effects of organic source concentration found as decisive factor on \\u003cem\\u003eLactobacillus rhamnosus\\u003c/em\\u003e BNC yield with 95% confidence interval. Field emission scanning electron microscopy (FESEM), fourier transform infrared spectroscopy (FTIR), differential / thermogravimetric thermal analysis (DTG/TG) were utilised to evaluate the structure and characterization of BNC.\\u003c/p\\u003e\\n\\u003cp\\u003eBNC production by \\u003cem\\u003eLactobacillus rhamnosus\\u003c/em\\u003e, with its biocompatible and biodegradable properties, environmentally friendly and low-cost nanomaterials have been produced with the potential to be used in many biomedical applications such as wound dressing and drug coating material.\\u003c/p\\u003e\",\"manuscriptTitle\":\"Production of New Nano-Bacterial Cellulose with Lactobacillus rhamnosus by Using Whey Waste as Substrate with Optimization Taguchi Method, which has the potential to be used in many biomedical products\",\"msid\":\"\",\"msnumber\":\"\",\"nonDraftVersions\":[{\"code\":1,\"date\":\"2024-01-05 05:04:36\",\"doi\":\"10.21203/rs.3.rs-3828016/v1\",\"editorialEvents\":[{\"type\":\"communityComments\",\"content\":0}],\"status\":\"published\",\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"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\":\"f4d70126-d324-4fd9-b5da-84f302b658f2\",\"owner\":[],\"postedDate\":\"January 5th, 2024\",\"published\":true,\"recentEditorialEvents\":[],\"rejectedJournal\":[],\"revision\":\"\",\"amendment\":\"\",\"status\":\"posted\",\"subjectAreas\":[],\"tags\":[],\"updatedAt\":\"2024-01-21T17:59:32+00:00\",\"versionOfRecord\":[],\"versionCreatedAt\":\"2024-01-05 05:04:36\",\"video\":\"\",\"vorDoi\":\"\",\"vorDoiUrl\":\"\",\"workflowStages\":[]},\"version\":\"v1\",\"identity\":\"rs-3828016\",\"journalConfig\":\"researchsquare\"},\"__N_SSP\":true},\"page\":\"/article/[identity]/[[...version]]\",\"query\":{\"redirect\":\"/article/rs-3828016\",\"identity\":\"rs-3828016\",\"version\":[\"v1\"]},\"buildId\":\"qtupq5eGEP_6zYnWcrvyt\",\"isFallback\":false,\"isExperimentalCompile\":false,\"dynamicIds\":[84888],\"gssp\":true,\"scriptLoader\":[]}","source_license":"CC-BY-4.0","license_restricted":false}