提高乙酸耐受性酿酒酵母JJJ1突变株构建及转录组学分析

Construction of Saccharomyces cerevisiae JJJ1 mutant for acetic acid tolerance improvement and transcriptomics analysis

中文关键词:  酿酒酵母  转录组学  JJJ1  乙酸  基因编辑  分子机制

英文关键词:Saccharomyces cerevisiae  transcriptomics  JJJ1 mutant  acetic acid  gene editing  molecular mechanism

中文摘要:

      目的 构建JJJ1突变株以提高酿酒酵母在发酵过程中的乙酸耐受性, 提高发酵效率。方法 本研究采用CRISPR-Cas9基因编辑技术构建酿酒酵母JJJ1突变株, 检测突变对酿酒酵母菌株的乙酸耐受性影响, 基于转录组学分析突变株耐受乙酸的分子机制。结果 在含有5 g/L乙酸的液体培养基中, 酿酒酵母JJJ1存活率是野生型菌株的4.44倍, 发酵120 h后酿酒酵母突变株JJJ1的细胞生物量是野生型菌株的1.15倍, 酿酒酵母JJJ1的生长延滞期比野生型菌株缩短了30 h; 转录组学研究表明, 敲除JJJ1基因增强酿酒酵母的代谢、生物调节、膜流动性以及转运活性和电子转移活性, 减少酿酒酵母细胞生理过程、细胞连接和拟核功能以及细胞结合功能。结论 敲除JJJ1基因的酿酒酵母突变株通过提高能量代谢和氨基酸合成, 降低核糖体生物发生减少细胞生理活动, 增强酿酒酵母菌株乙酸耐受性。

英文摘要:

      Objective To construct Saccharomyces cerevisiae JJJ1 mutant, and improve acetic acid tolerance and fermentation efficiency of Saccharomyces cerevisiae during fermentation process. Methods CRISPR-Cas9 technology was used to construct Saccharomyces cerevisiae JJJ1 mutant. The effect of JJJ1 mutation on acetic acid tolerance of Saccharomyces cerevisiae was detected. The molecular mechanism of acetic acid tolerance of Saccharomyces cerevisiae JJJ1 was further explored based on the transcriptomics analysis. Results The survival rate of Saccharomyces cerevisiae JJJ1 was 4.44 times in comparison to that of the wild-type strain in liquid medium containing 5 g/L acetic acid. After 120 h of fermentation, the cell biomass of Saccharomyces cerevisiae JJJ1 was 1.15 times in comparison to that of the wild-type strain. The growth lag period time of Saccharomyces cerevisiae JJJ1 was 30 h shorter than that of the wild-type strain. Transcriptomics studies showed that the deletion of Saccharomyces cerevisiae JJJ1 enhanced the metabolism, biological regulation, membrane fluidity, transport activity and electron transfer activity, and reduced the physiological process of cells, cell junction, nucleoid function and cell binding function. Conclusion The deletion of JJJ1 gene can improve energy metabolism and amino acid synthesis and reduce ribosomal biogenesis and cell physiological activity in Saccharomyces cerevisiae.