过表达Trxyr1基因对里氏木霉合成纤维素酶的影响  

Effect of overexpression of Trxyr1 gene on cellulase synthesis by Trichoderma reesei

作　　者：刘文书 王赞丞 高玉杉 莫易 彭念 冉小琴 李勇昊 LIU Wenshu;WANG Zancheng;GAO Yushan;MO Yi;PENG Nian;RAN Xiaoqin;LI Yonghao(School of Chemistry and Chemical Engineering,Chongqing University of Science and Technology,Chongqing 401331,China;Chongqing Key Laboratory of Industrial Fermentation Microorganisms,Chongqing University of Science and Technology,Chongqing 401331,China)

机构地区：[1]重庆科技学院化学化工学院,重庆401331 [2]重庆科技学院工业发酵微生物重庆市重点实验室,重庆401331

出　　处：《中国酿造》2023年第8期58-64,共7页China Brewing

基　　金：国家自然科学基金青年基金项目(21808022);重庆市自然科学基金项目(CSTB2022NSCQ-MSX0544);重庆科技学院硕士研究生创新计划项目(YKJCX2220502)。

摘　　要：以里氏木霉(Trichoderma reesei)Rut C30为出发菌株,利用组成型启动子(PDC1)过表达同源Trxyr1基因,探究阳性转化子T.reesei OExyr1在不同诱导物及非诱导条件下合成纤维素酶的情况,最后粗酶液被用于水解碱预处理的玉米秸秆以分析其催化性能。结果表明,T.reesei OExyr1在葡萄糖-β二糖混合物诱导下纤维素酶活是野生型菌株的1.22倍;而以纤维素为诱导物,纤维素酶活可提高2.95倍;此外在非诱导条件下T.reesei OExyr1可合成纤维素酶;但T.reesei OExyr1所产纤维素酶对玉米秸秆的水解效率与野生型无显著性差异。以上结果说明不同诱导物下Trxyr1基因高表达均可提升纤维素酶产量,尤其对于固体纤维素诱导物,但Trxyr1基因表达量的提高对纤维素酶系影响不大。研究成果对阐明T.reesei合成纤维素酶的分子机制及构建高产纤维素酶菌株提供参考。Using Trichoderma reesei Rut C30 as the starting strain,and homologous Trxyr1 gene was overexpressed by constitutive promoter(PDC1).The synthesis of cellulase by the positive transformer T.reesei OExyr1 under different inductors and non-inducible conditions were investigated.Finally,alkali-pretreated corn stalk was hydrolyzed by the crude enzyme liquid to analyze its catalytic performance.The results showed that the cellulase activity of T.reesei OExyr1 was 1.22 times that of the wild-type strain induced by glucose-β-bisaccharide mixture.With cellulose as the inducer,cellulase activity increased by 2.95 times.In addition,T.reesei OExyr1 could synthesis cellulase under non-induced conditions.However,the hydrolysis efficiency of cellulase produced by T.reesei OExyr1 on corn stalk was not significantly different from that of wild type.These results indicated that high expression of Trxyr1 gene could increase cellulase production under different inducers,especially for solid cellulose inducers,but the increase of Trxyr1 gene expression had little effect on cellulase series.The research results provided reference for elucidating the molecular mechanism of cellulase synthesis by T.reesei and constructing high-yielding cellulase strains.

关 键 词：里氏木霉 纤维素酶 诱导物 Trxyr1基因 转录因子 玉米秸秆 

分 类 号：Q81[生物学—生物工程] TS210.9[轻工技术与工程—粮食、油脂及植物蛋白工程]