利用易错PCR构建酱油酿造用T酵母spt15突变库  

Construction of Mutant Library of spt15 of Candida versatilis by Error-Prone PCR

作　　者：赵秀丽[1] 张毓秀 孙月芹 侯丽华[1] ZHAO Xiuli;ZHANG Yuxiu;SUN Yueqin;HOU Lihua(Institute for New Rural Development,Tianjin University of Science and Technology,Tianjin 300457,Chin)

机构地区：[1]天津科技大学新农村发展研究院,天津300457

出　　处：《食品科学》2018年第14期106-111,共6页Food Science

基　　金：国家自然科学基金面上项目(31371819);天津市科技支撑重点项目(17YFZCNC00460);天津科技大学新农村发展研究院开放基金资助项目(xnc201604);天津市高等学校创新团队培养计划资助项目(TD13-5013)

摘　　要：为得到酱油酿造优良性状的微生物,对酱油酿造用T酵母进行研究。首先利用易错聚合酶链式反应(polymerase chain reaction,PCR)技术构建spt15(可编码TATA结合蛋白)的突变基因库,然后重组于表达载体YEplac195中,并导入尿嘧啶营养缺陷型菌株W303和WM2中筛选。结果表明:在不同盐质量分数的固体培养基中经筛选获得4个最佳突变菌株,其具有耐盐优势,在酱油发酵中可产生较高的氨基酸态氮。经PCR产物测序可知,重组质粒中目的基因的序列有碱基的增添、缺失和替换。The objective of the present study was to develop mutant strains of Candida versatilis for the production of soy sauce. A mutant library of spt15 encoding the TATA-binding protein of C. versatilis was constructed by error-prone PCR and it was recombined into the expression vector YEplac195 and then introduced into uracil auxotrophic strains W303 and WM2 for screening purpose. The results showed that the four best mutant strains A1, A2, A3, and A4 were obtained through screening in solid medium with different salt concentrations. Compared with the control group, their salt tolerance was improved and soy sauces fermented by these mutant strains contained higher contents of amino nitrogen. The sequence of the recombinant plasmid showed base insertion, deletion and substitution as indicated by the sequencing of PCR products.

关 键 词：易错PCR spt15 基因突变库 筛菌 酱油发酵 

分 类 号：TS201.3[轻工技术与工程—食品科学]