摘要
为提高乳清酸到尿嘧啶核苷酸(UMP)的转化效率,利用PCR方法扩增酿酒酵母乳清酸磷酸核糖转移酶基因URA5,并将其连接到携带乳清苷酸脱羧酶基因URA3的表达载体pYX212中,构建了重组质粒pYX212-URA5,然后转化到酿酒酵母BJX12中进行表达,并进行转化乳清酸到UMP的初步研究。试验结果表明:pYX212-URA5/BJX12发酵培养40h后以32mmol/L乳清酸为底物催化产生UMP的量约为7mmol/L。明显高于同等条件下pYX212/BJX12的UMP产量2.7mmol/L和对照组野生型BJX12的UMP产量2.4mmol/L。
To increase the biotransfomation efficiency from the orotic acid to the uridine 5＇ -monophosphate （UMP）, URA5 gene encoding orotate phosphoribosytransferase was amplified from Saccharomyces cerevisiae BY4742 by PCR, then it was inserted into the expression vector pYX212（ contained orotidine monophosphate decarboxylase gene URA3 ）and the pYX212-URA5 was transformed into Saccharomyces cerevisiae BJX12 by electroporation. The recombinant strain was elementarily used to convert orotic acid to UMP. The results showed that pYX212-URA5/BJX12 could accumulate 7mmol/L UMP from 32mmol/L orotic acid in 26h, significantly higher than both control groups pYX212/BJX12 （2.7mmol/L） and BJX12（2.4 mmoL/L）.
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2008年第12期77-81,共5页
China Biotechnology
基金
国家"863"计划(2006AA02Z236)
江苏省自然科学基金(BK2007527)资助项目
关键词
乳清酸磷酸核糖转移酶
乳清苷酸脱羧酶
酿酒酵母
生物催化
尿嘧啶核苷酸
Orotate phosphoribosytransferase （ OPRTase ） Biocatalysis Saccharomyces cerevisiae Uridine 5＇ -monophosphate （UMP） Orotidine monophosphate decarboxylase （ODCase）
