UdhA和博伊丁假丝酵母xylI基因共表达对木糖醇发酵的影响  

The Effect of Co-expression of UdhA and Candida boidinii xylI Gene on Xylitol Fermentation

作　　者：唐梅 蔡松 付声亮 王金华[1] 王永泽[1] TANG Mei;CAI Song;FU Sheng-liang(Hubei Key Laboratory of Industrial Microbiology,Hubei University of Technology,Wuhan,Hubei 430068)

机构地区：[1]湖北工业大学工业微生物教育部重点实验室,湖北武汉430068

出　　处：《安徽农业科学》2022年第1期106-109,共4页Journal of Anhui Agricultural Sciences

基　　金：国家“十二五”支撑计划项目(2012BAD27B03);山东创新计划项目(201720311004)。

摘　　要：[目的]研究可溶性吡啶核苷酸转氢酶基因(UdhA)和醛糖还原酶基因(xylI)共表达对木糖醇发酵的影响。[方法]将来源于博伊丁假丝酵母醛糖还原酶xylI基因克隆到pET28a(+)上,并在BL21(DE3)中表达,通过SDS-PAGE对表达产物的分子量和酶活进行测定。随即将xylI基因连接lacP启动子,构建pWYZ-2质粒并将其转化到E.coli AI07菌株。进一步将来源于E.coli W3110的UdhA基因克隆到pWYZ-2质粒实现与xylI共表达,所构建pWYZ-4质粒转化到E.coli AI07菌株,比较了E.coli AI07/pWYZ-2和E.coli AI07/pWYZ-4木糖醇发酵结果。[结果]在BL21(DE3)/pET28a(+)体系诱导表达的醛糖还原酶分子量为39 kD,木糖还原酶酶活为3.30 U/mL。E.coli AI07/pWYZ-2菌株发酵48 h,木糖醇产量为19.90 g/L;E.coli AI07/pWYZ-4发酵36 h,木糖醇产量达到19.91 g/L,较菌株AI07/pWYZ-2生产强度提高了33.25%。[结论]通过将UdhA基因与xylI基因进行共表达,提高了重组大肠杆菌(E.coli AI07/pWYZ-4)合成木糖醇的生产强度。[Objective]The effect of co-expression of soluble pyridine nucleotide transhydrogenase gene(UdhA)and aldose reductase gene(xylI)on xylitol fermentation was investigated.[Method]xylI gene from Candida boidinii encoding aldose reductase was cloned into pET28a(+)and expressed in BL21(DE3).The enzyme molecular weight was detected by SDS-PAGE and enzyme activity was assayed.xylI gene was ligated to the lacP promoter for construction of plasmid pWYZ-2 and then the plasmid was transformed into E.coli AI07.The UdhA gene derived from E.coli W3110 was further cloned into pWYZ-2 plasmid to achieve co-expression with xylI for pWYZ-4 plasmid construction and then pWYZ-4 was transformed into E.coli AI07.E.coli AI07/pWYZ-2 was compared with E.coli AI07/pWYZ-4 for xylitol fermentation.[Result]The molecular weight of xylose reductase expressed in BL21(DE3)/pET28a(+)system was 39 kD,and the enzymatic activity of aldose reductase was 3.3 U/mL.The E.coli AI07/pWYZ-2 strain was fermented for 48 hours,the xylitol output was 19.90 g/L.E.coli AI07/pWYZ-4 was fermented for 36 hours,xylitol production reached 19.91 g/L,xylitol productivity of E.coli AI07/pWYZ-4 was 33.25%higher than that of strain E.coli AI07/pWYZ-2.[Conclusion]Xylitol productivity of recombinant Escherichia coli was improved using co-expression of UdhA gene and xylI gene.

关 键 词：博伊丁假丝酵母 xylI基因 UdhA基因 共表达 木糖醇 

分 类 号：Q812[生物学—生物工程]