摘要
首次从丙酮丁醇梭菌(Clostridium acetobutylicum ATCC824)中克隆得到L-乳酸脱氢酶(L-lactate dehydrogenase,ldhL)基因,并将其连接到pSE380表达载体上,得到重组质粒pSE380ldhL,将重组质粒转化到乳酸脱氢酶和丙酮酸裂解酶缺陷的Escherichia coli FMJl44大肠杆菌中进行表达。SDS-PAGE分析表达产物的分子量约为34kD,摇瓶发酵后用HPLC检测分析L-乳酸产量为2.4g/L,纯度达到99.9%,不需要再进行手性分离,为以后在工业上生物法生产高纯度的L-乳酸打下基础。
The L-lactate dehydrogenase gene was firstly cloned from Clostridium acetobutylicum ATCC 824. The recombinant plas- mid pSE3801dhL was constructed by inserting ldhL into expression vector pSE380 and then transformed into Escherichia coli FMJI44. SDS-PAGE result showed that the relative molecular weight of the expression product was about 34 kD. The production of L-Lactic acid analyzed by HPLC was about 2.4 g/L after fermentation in the flask. The degree of purity was 99. 9% ,which does not require Chiral separation further. The research lay a foundation to achieve biological production of high-purity with high purity in the industry area.
出处
《生物技术通报》
CAS
CSCD
北大核心
2010年第3期81-84,89,共5页
Biotechnology Bulletin
基金
广西青年基金(桂科青0832016)
广西亚热带生物资源保护利用重点实验室--省部共建国家重点实验室培育基地开放课题(SB0606)
关键词
L-乳酸
L-乳酸脱氢酶
克隆
表达
L-Lactic acid L-lactate dehydrogenase Cloning Expression
