基于等离子体诱变提高木糖利用重组酿酒酵母的抑制物耐受性  

Improvement of inhibitor tolerance of xylose-fermenting recombinant Saccharomyces cerevisiae via atmospheric and room temperature plasma mutagenesis

作　　者：陈栋 吴娅菁 缪晡 汤岳琴[2,3] CHEN Dong;WU Yajing;MIAO Bu;TANG Yueqin(Sinopec Shanghai Engineering Co.Ltd.,Shanghai 200120,China;Environmental Biotechnology Research Center,College of Architecture and Environment,Sichuan University,Chengdu 610065,China;Sichuan Environmental Protection Key Laboratory of Organic Waste Resource Utilization,Chengdu 610065,China)

机构地区：[1]中石化上海工程有限公司,上海200120 [2]四川大学建筑与环境学院环境生物技术研究中心,成都610065 [3]四川省环境保护有机废弃物资源化利用重点实验室,成都610065

出　　处：《应用与环境生物学报》2021年第6期1464-1470,共7页Chinese Journal of Applied and Environmental Biology

基　　金：国家重点研发计划重点专项(2019YFE012953);国家自然科学基金项目(31170093)资助。

摘　　要：木质纤维素原料预处理过程中产生的各种抑制物是燃料乙醇生产的一大障碍,因此要求酿酒酵母菌株具有优良的抑制物耐受能力.相比葡萄糖发酵,酿酒酵母利用木糖的发酵过程对抑制物更敏感.针对一株具有良好木糖发酵能力的絮凝性工业重组酿酒酵母菌株s6,进行常压室温等离子体(Atmospheric and Room Temperature Plasma,ARTP)诱变,通过平板初筛和发酵复筛,获得一株抑制物耐受性得到显著提升的突变菌株335.与出发菌株s6相比,在含单一抑制物(4.8 g/L乙酸,1.9 g/L 5-羟甲基糠醛,0.46 g/L香草醛)的发酵条件下,突变菌株335利用木糖发酵时的乙醇浓度分别提高了55.0%、13.7%和35.0%,在含混合抑制物(2.4 g/L乙酸+1.3 g/L 5-羟甲基糠醛+0.23 g/L香草醛)的发酵条件下的乙醇浓度提高了37.2%.突变菌株335能够利用含高浓度抑制物的秸秆水解液进行发酵,表现出优良的抑制物耐受能力.本研究结果表明ARTP诱变是获得优良抑制物耐受突变菌株的一种有效手段,突变菌株335对单一抑制物以及混合抑制物均具有较好的耐受能力,具有工业应用潜力.Inhibitors produced during the pretreatment process of ethanol fermentation are a great challenge to lignocellulosic ethanol production, which requires excellent stress tolerance in Saccharomyces cerevisiae strains. Compared with glucose fermentation, xylose fermentation by S. cerevisiae is much more sensitive to these inhibitors. In this study, atmospheric and room temperature plasma(ARTP) mutagenesis was performed on a xylose-fermenting flocculating industrial recombinant S. cerevisiae strain(s6), and a mutant strain(335).The mutant strain showed significantly improved stress tolerance and was screened out by plate screening and fermentation assessment. During xylose fermentation, the ethanol concentration of mutant strain 335 increased by 55.0%, 13.7%, and 35.0% compared with the original strain s6, with the addition of single inhibitors 4.8 g/L acetic acid, 1.9 g/L 5-hydroxymethylfurfural, and 0.46 g/L vanillin, respectively. Ethanol concentration increased by 37.2%with the addition of mixed inhibitors 2.4 g/L acetic acid, 1.3 g/L 5-hydroxymethylfurfural and 0.23 g/L vanillin. The mutant strain 335 presented satisfactory fermentation performance when using straw hydrolysate containing high concentrations of inhibitors, suggesting its strong resistance to mixed inhibitors. The results indicated that ARTP mutagenesis is effective for achieving stress-tolerant mutants, and that the mutant strain 335 obtained in the present study has potential for industrial application, due to its strong tolerance to mixed inhibitors.

关 键 词：纤维素燃料乙醇 工业酿酒酵母 抑制物耐受性 木糖发酵 常压室温等离子体诱变 

分 类 号：TQ223.122[化学工程—有机化工] TQ920.1[轻工技术与工程—发酵工程]