Rice bran (RB) and de-oiled rice bran (DRB) have been treated and used as the carbon source in acetone-butanol-ethanol (ABE) production using Clostridium saccharoperbutylacetonicum N1-4. The results showed that pretreated DRB produced more ABE than pretreated RB. Dilute sulfuric acid was the most suitable treatment method among the various pretreatment methods that were applied. The highest ABE obtained was 12.13 g/L, including 7.72 g/L of biobutanol, from sulfuric acid. The enzymatic hydrolysate of DRB (ESADRB), when treated with XAD-4 resin, resulted in an ABE productivity and yield of 0.1 g/L h and 0.44 g/g, respectively. The results also showed that the choice of pretreatment method for RB and DRB is an important factor in butanol production.
Continuous fermentation of dilute acid-pretreated de-oiled rice bran (DRB) to butanol by the Clostridium acetobutylicum YM1 strain was investigated. Pretreatment of DRB with dilute sulfuric acid (1%) resulted in the production of 42.12 g/L total sugars, including 25.57 g/L glucose, 15.1 g/L xylose and 1.46 g/L cellobiose. Pretreated-DRB (SADRB) was used as a fermentation medium at various dilution rates, and a dilution rate of 0.02 h−1 was optimal for solvent production, in which 11.18 g/L of total solvent was produced (acetone 4.37 g/L, butanol 5.89 g/L and ethanol 0.92 g/L). Detoxification of SADRB with activated charcoal resulted in the high removal of fermentation inhibitory compounds. Fermentation of detoxified-SADRB in continuous fermentation with a dilution rate of 0.02 h−1 achieved higher concentrations of solvent (12.42 g/L) and butanol (6.87 g/L), respectively, with a solvent productivity of 0.248 g/L.h. This study showed that the solvent concentration and productivity in continuous fermentation from SADRB was higher than that obtained from batch culture fermentation. This study also provides an economic assessment for butanol production in continuous fermentation process from DRB to validate the commercial viability of this process.
The effect of pH and butyric acid supplementation on the production of butanol by a new local isolate of YM1 during batch culture fermentation was investigated. The results showed that pH had a significant effect on bacterial growth and butanol yield and productivity. The optimal initial pH that maximized butanol production was pH 6.0 ± 0.2. Controlled pH was found to be unsuitable for butanol production in strain YM1, while the uncontrolled pH condition with an initial pH of 6.0 ± 0.2 was suitable for bacterial growth, butanol yield and productivity. The maximum butanol concentration of 13.5 ± 1.42 g/L was obtained from cultures grown under the uncontrolled pH condition, resulting in a butanol yield ( ) and productivity of 0.27 g/g and 0.188 g/L h, respectively. Supplementation of the pH-controlled cultures with 4.0 g/L butyric acid did not improve butanol production; however, supplementation of the uncontrolled pH cultures resulted in high butanol concentrations, yield and productivity (16.50 ± 0.8 g/L, 0.345 g/g and 0.163 g/L h, respectively). pH influenced the activity of NADH-dependent butanol dehydrogenase, with the highest activity obtained under the uncontrolled pH condition. This study revealed that pH is a very important factor in butanol fermentation by YM1.
The interaction between sucrose, yeast extract and initial pH was investigated to optimize critical medium components for mycelium biomass and production of exopolysaccharide (EPS) of Lentinus squarrosulus using Response Surface Methodology (RSM). A central composite design (CCD) was applied and a polynomial regression model with quadratic term was used to analyse the experimental data using analysis of variance (ANOVA). ANOVA analysis showed that the model was very significant (p < 0.0001) for both mycelium biomass and EPS production. The yeast extract concentrations and initial pH showed the strongest effect (p < 0.001) meanwhile sucrose concentrations showed significant value at p < 0.005 for mycelium biomass. For EPS production, only sucrose and yeast extract concentrations gave significant value with p < 0.001 and p < 0.005 respectively. The model was validated by applying the optimized conditions and 25.8 ± 0.9 g/L of mycelium biomass, and 5.7 ± 0.4 mg/mL EPS concentration was obtained. The estimated optimum conditions of the variables for the production of mycelium biomass and EPS production by Lentinus squarrosulus are as follows: sucrose concentration 114.61 g/L, yeast extract 1.62 g/L and initial pH of 5.81; sucrose concentration 115.8 g/L, yeast extract of 3.39 g/L and initial pH of 6.44 respectively.
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