乙醇作为一种可再生能源,其生产主要以酵母代谢为主,而筛选高耐性高产乙醇菌株是乙醇产率高的关键。该文以6株现有单倍体酿酒酵母为出发菌株,通过常温常压等离子诱变构建突变文库,经氯化三苯基四氮唑显色初筛,40、41 ℃压力生长复筛,发酵筛选培养基定量分析。最终获得9株优势突变酿酒酵母单倍体菌株,经不同配型单倍体杂交,成功获得一株发酵效率高,耐温、耐酸、耐乙醇性能均优于亲本菌株的二倍体酿酒酵母菌株C6。结果显示,C6菌株35 ℃复合糖水发酵培养基发酵终点酒精度为14.6%,亲本菌株AE2和M则为12.82%和13.72%,酒精度分别提高13.8%和6.4%,C6菌株在30 ℃条件下,酒精度可达17.9%。耐温性能提高较为明显,40 ℃条件下生长,C6、AE2和M菌株平稳期OD680值分别为4.12、0.35和1.15,生物量增幅在1.2～12倍。C6菌株在乙醇生产领域具有较高的工业应用价值。
Ethanol, a renewable energy, relies predominantly on yeast metabolism for its production.Selecting strains exhibiting high tolerance and productivity in ethanol fermentation is the key to achieve enhanced ethanol yields.In order to select and breed high-tolerant and high-yielding ethanol yeast strains, with six existing haploid brewing yeast strains, a mutant library was constructed through atmospheric and room temperature plasma mutagenesis, which were colorimetrically screened by triphenyl tetrazolium chloride, re-screened by pressure growth at 40, 41 ℃, and quantitatively analyzed by fermentation screening medium.Subsequently, nine dominant mutant haploid strains were obtained and hybridized through different mating types, resulting in a diploid brewing yeast strain, C6.with superior fermentation efficiency, better temperature resilience, acidity tolerance, and ethanol resistance than parental strains.The results exhibited that in a 35 ℃ composite sugar water fermentation medium, the C6 strain achieved an ethanol content of 14.6%, whereas parental strains AE2 and M reached 12.82% and 13.72%, respectively.Notably, the ethanol content increased by 13.8% and 6.4%, while at 30 ℃, the ethanol content for C6 came to 17.9%.It was evident that the stable OD680 values period of the improved temperature tolerance of C6, AE2, and M strains were 4.12, 0.35, 1.15, respectively, under 40 ℃ conditions, showing a biomass increase ranging from 1.2 to 12 times.Thus, the C6 strain exhibits promising industrial applications in the field of ethanol production
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