Hu Meng-Long, Zha Jian, He Lin-Wei, Lv Ya-Jin, Shen Ming-Hua, Zhong Cheng, Li Bing-Zhi, Yuan Ying-Jin
Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin UniversityTianjin, China; Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), Tianjin UniversityTianjin, China.
Key Laboratory of Industrial Fermentation Microbiology (Ministry of Education), Tianjin University of Science and Technology Tianjin, China.
Front Microbiol. 2016 Mar 3;7:241. doi: 10.3389/fmicb.2016.00241. eCollection 2016.
Cellobiose accumulation and the compromised temperature for yeast fermentation are the main limiting factors of enzymatic hydrolysis process during simultaneous saccharification and fermentation (SSF). In this study, genes encoding cellobiose transporter and β-glucosidase were introduced into an industrial Saccharomyces cerevisiae strain, and evolution engineering was carried out to improve the cellobiose utilization of the engineered yeast strain. The evolved strain exhibited significantly higher cellobiose consumption rate (2.8-fold) and ethanol productivity (4.9-fold) compared with its parent strain. Besides, the evolved strain showed a high cellobiose consumption rate of 3.67 g/L/h at 34°C and 3.04 g/L/h at 38°C. Moreover, little cellobiose was accumulated during SSF of Avicel using the evolved strain at 38°C, and the ethanol yield from Avicel increased by 23% from 0.34 to 0.42 g ethanol/g cellulose. Overexpression of the genes encoding cellobiose transporter and β-glucosidase accelerated cellobiose utilization, and the improvement depended on the strain background. The results proved that fast cellobiose utilization enhanced ethanol production by reducing cellobiose accumulation during SSF at high temperature.
纤维二糖积累和酵母发酵的温度受限是同步糖化发酵(SSF)过程中酶水解的主要限制因素。本研究将编码纤维二糖转运蛋白和β-葡萄糖苷酶的基因导入工业酿酒酵母菌株,并进行进化工程以提高工程酵母菌株对纤维二糖的利用。与亲本菌株相比,进化菌株的纤维二糖消耗速率显著提高(2.8倍),乙醇生产率提高(4.9倍)。此外,进化菌株在34°C时纤维二糖消耗速率为3.67 g/L/h,在38°C时为3.04 g/L/h。而且,在38°C使用进化菌株对微晶纤维素进行SSF时,几乎没有纤维二糖积累,微晶纤维素的乙醇产率从0.34 g乙醇/g纤维素提高到0.42 g乙醇/g纤维素,提高了23%。编码纤维二糖转运蛋白和β-葡萄糖苷酶的基因的过表达加速了纤维二糖的利用,且这种改善依赖于菌株背景。结果证明,快速的纤维二糖利用通过减少高温下SSF过程中的纤维二糖积累提高了乙醇产量。