Purpose Yeasts are widely used for the production of bioethanol from biomasses rich in sugar. The present study was aimed at isolating, screening, and characterizing fermentative wild yeast recovered from bio-waste and co-products of Ethiopian sugar factories for bioethanol production using sugarcane molasses as a substrate. Method The wild yeasts were identified according to their cellular morphology and D1/D2 and ITS1-5.8S-ITS2 rDNA sequencing. Analysis of ethanol and by-product concentration was done by HPLC equipped with a UV detector. Higher alcohols, acetaldehyde, and methanol were analyzed using GC-MS equipped with a flame ionization detector (FID). Result Seven strains (Meyerozyma caribbica MJTm3, Meyerozyma caribbica MJTPm4, Meyerozyma caribbica SHJF, Saccharomyces cerevisiae TA2, Wickerhamomyces anomalus MJTPm2, Wickerhamomyces anomalus 4m10, and Wickerhamomyces anomalus HCJ2F) were found tolerant to 18% (v/v) ethanol, whereas one strain Meyerozyma caribbica MJTm3 tolerated 20%. These strains also showed tolerance to 45°C, 50% of sugar, and pH 2–10. Meyerozyma caribbica MJTm3 produced 12.7% (v/v) of alcohol with an actual ethanol concentration of 26 g L−1, an ethanol yield of 47%, 78% of theoretical yield, and a productivity of 0.54 g L−1 h−1 from 30 °Brix of molasses at 48 h incubation under laboratory scale. Based on the one variable at a time optimization (OVAT), the optimal parameters for maximum bioethanol production were at initial pH 5.5, 35 °Brix, 30°C, 15% inoculum size, 150 rpm, 4 g L−1 di-ammonium phosphate supplement, and 48 h incubation. Under these optimum conditions, 14% (v/v) alcohol, 42 g L−1 actual ethanol concentration, 69% ethanol yield, 89% of theoretical yield, and productivity of 0.88 g L−1 h−1 were obtained. Conclusion These results indicated that M. caribbica MJTm3 should further be evaluated, optimized, and improved for industrial bioethanol production due to its fermentation potential.
Purpose Co-culturing of stress-tolerant fermenting yeasts is a widely used method to improve bioethanol production from biomass enriched in fermentable sugars. This study aims to produce bioethanol from sugarcane molasses by simultaneous co-fermentation of S. cerevisiae isolate TA2 and W. anomalus isolate HCJ2F-19. Method Response surface methodology (RSM) based on the central composite design (CCD) was employed to optimize fermentation conditions, including mixing rate (110–150 rpm), temperature (25–35 oC), molasses concentration (25–35 obrix), and incubation time (36–72 h). The ethanol concentration was analyzed using HPLC equipped with a UV detector. Results The mono-culture, S. cerevisiae TA2 produces 17.2 g.L− 1 of ethanol, 0.33 g.g− 1 of ethanol yield, and 0.36 g.L− 1.h− 1 of productivity compared to W. anomalus HCJ2F which produces 14.5 g.L− 1, 0.30 g.g− 1 and 0.28 g.L− 1.h− 1 ethanol, ethanol yield, and productivity under laboratory conditions, respectively. In comparison to single cultures of S. cerevisiae TA2, and W. anomalus HCJ2F, the co-fermentation showed an increased ethanol yield of 29% and 53% compared to the single species fermentations, respectively. The results showed that the growth of W. anomalus HCJ2F-19 and S. cerevisiae TA2 was not influenced by each other during the co-fermentation process. The one variable at a time optimization (OVAT) demonstrated an ethanol concentration of 26.5 g.L− 1 with a specific yield and productivity of 0.46 g.g− 1, 0.55 g.L− 1.h− 1, respectively, at pH 5.5, 25 obrix, 48 h, 150 rpm, 30oC, 60:40 inoculum ratio, and 10% overall inoculum size. The maximum ethanol concentration of 35.5 g.L− 1 was obtained by co-fermentation using the RSM-CCD tool at 30 obrix, 30oC, 54 h, and 130 rpm. Conclusion The results suggested that the co-fermentation of S. cerevisiae TA2 and W. anomalus HCJ2F improves bioethanol production under optimum fermentation conditions.
Purpose Yeast strains tolerant to a wide range of stress conditions are needed for the production of bioethanol from substrates rich in sugar. In our earlier research findings, Meyerozyma caribbica isolate MJTm3 (OM329077) demonstrated remarkable stress tolerance and fermentative activity. The present study aimed to optimize six fermentation parameters to generate conducive fermentation conditions for ethanol production by M. caribbica isolate MJTm3. Method The response surface method (RSM) based on central composite design (CCD) was employed to optimize process conditions for higher bioethanol yield. The optimization process was carried out based on six independent parameters, namely temperature (25–35 °C), pH (5.5–6.5), inoculum size (10–20% (v/v)), molasses concentration (25–35 (w/v)), mixing rate (110–150 rpm), and incubation period (48–72-h). Analysis of ethanol concentration was done by HPLC equipped with a UV detector. Result The optimal conditions of the parameters resulting in a maximum predicted ethanol yield were as follows: pH 5.5, an inoculum size of 20%, a molasses concentration of 25 °Bx, a temperature of 30 °C, an incubation period of 72-h, and a mixing rate of 160 revolutions per minute (rpm). Using the above optimum conditions, the model predicted a bioethanol yield of 79%, 92% of the theoretical yield, a bioethanol concentration of 49 g L−1, and a productivity of 0.68 g L−1 h−1. A batch fermentation experiment was carried out to validate the predicted values and resulted in a bioethanol yield of 86%, 95% of theoretical yield, a bioethanol concentration of 56 g L−1, and productivity of 0.78 g L−1 h−1. On the other hand, the surface plot analysis revealed that the synergistic effect of the molasses concentration and the mixing rate were vital to achieving the highest bioethanol yield. These values suggested that the RSM with CCD was an effective method in producing the highest possible output of bioethanol from molasses in actual operation. Conclusion The study confirmed the potential of using M. caribbica isolate MJTm3 for bioethanol production from sugarcane molasses under the abovementioned optimal fermentation conditions.
Purpose Yeast strains tolerant to a wide range of stress conditions are needed for the production of bioethanol from substrates rich in sugar. In our earlier research findings, Meyerozyma caribbica isolate MJTm3 (OM329077) demonstrated remarkable stress tolerance and fermentative activity. The present study aimed to optimize six fermentation parameters to generate conducive fermentation conditions for ethanol production by M. caribbica isolate MJTm3. Method The response surface method (RSM) based on central composite design (CCD) was employed to optimize process conditions for higher bioethanol yield. The optimization process was carried out based on six independent parameters, namely temperature (25-30oC), pH (5.5–6.5), inoculum size (10–20% (v/v)), molasses concentration (25–35 (w/v)), mixing rate (110–150 rpm), and incubation period (48–72 h). Analysis of ethanol concentration was done by HPLC equipped with a UV detector. Result The optimal conditions of the parameters resulting in a maximum predicted ethanol yield were as follows: pH 5.5, an inoculum size of 20%, a molasses concentration of 25 oBrix, a temperature of 30oC, an incubation period of 72 h, and a mixing rate of 160 revolutions per minute (rpm). Using the above optimum conditions, the model predicted a bioethanol yield of 79%, 92% of the theoretical yield, a bioethanol concentration of 49 g L− 1, and a productivity of 0.68 g L− 1h− 1. A batch fermentation experiment was carried out to validate the predicted values and resulted in a bioethanol yield of 86%, 95% of theoretical yield, a bioethanol concentration of 56 g L− 1, and productivity of 0.78 g L− 1h− 1. On the other hand, the surface plot analysis revealed that the synergistic effect of the molasses concentration and the mixing rate were vital to achieving the highest bioethanol yield. In conclusion, these values suggested that the RSM with CCD was an effective method in producing the highest possible output of bioethanol from molasses in actual operation. Conclusion The study confirmed the potential of using M. caribbica isolate MJTm3 for bioethanol production from sugarcane molasses under the above-mentioned optimal conditions.
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