Seong Yeong-Je, Park Haeseong, Yang Jungwoo, Kim Soo-Jung, Choi Wonja, Kim Kyoung Heon, Park Yong-Cheol
Department of Bio and Fermentation Convergence Technology, and BK21 PLUS Program, Kookmin University, Seoul, 136-702, South Korea.
Department of Biotechnology, Graduate School, Korea University, Seoul, 136-713, South Korea.
Appl Microbiol Biotechnol. 2017 May;101(9):3567-3575. doi: 10.1007/s00253-017-8139-2. Epub 2017 Feb 6.
The SPT15 gene encodes a Saccharomyces cerevisiae TATA-binding protein, which is able to globally control the transcription levels of various metabolic and regulatory genes. In this study, a SPT15 gene mutant (S42N, S78R, S163P, and I212N) was expressed in S. cerevisiae BY4741 (BSPT15-M3), of which effects on fermentative yeast properties were evaluated in a series of culture types. By applying different nitrogen sources and air supply conditions in batch culture, organic nitrogen sources and microaerobic condition were decided to be more favorable for both cell growth and ethanol production of the BSPT15-M3 strain than the control S. cerevisiae BY4741 strain expressing the SPT15 gene (BSPT15wt). Microaerobic fed-batch cultures of BSPT15-M3 with glucose shock in the presence of high ethanol content resulted in a 9.5-13.4% higher glucose consumption rate and ethanol productivity than those for the BSPT15wt strain. In addition, BSPT15-M3 showed 4.5 and 3.9% increases in ethanol productivity from cassava hydrolysates and corn starch in simultaneous saccharification and fermentation processes, respectively. It was concluded that overexpression of the mutated SPT15 gene would be a potent strategy to develop robust S. cerevisiae strains with enhanced cell growth and ethanol production abilities.
SPT15基因编码一种酿酒酵母TATA结合蛋白,它能够全局控制各种代谢和调控基因的转录水平。在本研究中,一个SPT15基因突变体(S42N、S78R、S163P和I212N)在酿酒酵母BY4741(BSPT15-M3)中表达,在一系列培养类型中评估了其对发酵酵母特性的影响。通过在分批培养中应用不同的氮源和供气条件,发现有机氮源和微需氧条件比表达SPT15基因的对照酿酒酵母BY4741菌株(BSPT15wt)更有利于BSPT15-M3菌株的细胞生长和乙醇生产。在高乙醇含量下对BSPT15-M3进行葡萄糖冲击的微需氧补料分批培养,其葡萄糖消耗率和乙醇生产率比BSPT15wt菌株分别高9.5 - 13.4%。此外,在同步糖化发酵过程中,BSPT15-M3从木薯水解物和玉米淀粉中产生乙醇的生产率分别提高了4.5%和3.9%。得出的结论是,突变的SPT15基因的过表达将是开发具有增强细胞生长和乙醇生产能力的健壮酿酒酵母菌株的有效策略。