%0 Journal Article
%T Expression and characterization of formate dehydrogenase gene in Klebisella pneumoniae<br>甲酸脱氢酶在Klebsiella pneumoniae中的表达和功能分析
%A HUANG Zhi-hu
%A ZHANG Yan-ping
%A LIU Ming
%A WANG Bao-guang
%A CAO Zhu-an
%A <br>黄志华
%A 张延平
%A 刘铭
%A 王宝光
%A 曹竹安
%J 微生物学报
%D 2007
%I 
%X Glycerol can be converted to 1,3-propanediol by the anaerobic fermentation of Klebsiella pneumoniae, during which reducing equivalent NADH was consumed. Therefore, the availability of NADH would be critical for the yield of 1, 3-propanediol. In this paper, formate/formate dehydrogenase system was used for the regeneration of in vivo NADH and the improvement of 1, 3-propanediol production. Formate Dehydrogenase gene (fdh) was amplified from Candida boidinii genome by PCR and the purified PCR product was inserted into the vector pMD18-T Simple to construct plasmid pMD18-T Simple-fdh, which was transformed into Escherichia coli DH5alpha and recombinants were selected by blue-white selection. From the transformant the fdh gene was separated and inserted into pMALTM-p2X to construct expression vector pMAL-p2X-fdh, which was transformed into Klebsiella pneumoniae YMU2 and a recombinant strain Klebsiella pneumoniae F-l was obtained. The plasmid stability of strain F-l and the conditions of fdh expression induced by IPTG were studied. It was demonstrated that the plasmid had good stability, and 0.5mmol/L IPTG would induce the expression of protein encoded by fdh gene with the molecular weight of 40.2kDa. The enzyme activity reached 5.47U/mg crude protein when K. pneumoniae F-I was induced for 4h by 0.5mmol/L IPTG. Compared with that of the parent strain K. pneumoniae YMU2, the yield of 1,3-propanediol of recombinant strain F-1 increased by 12.5% in the anaeribic bioreactor.
%K Candida boidinii
%K Klebsiella pneumoniae
%K NADH<br>甲酸脱氢酶
%K 1
%K 3-丙二醇
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A3C6BA55AB623B90FA9104CFFC826F3C&aid=AB352B826265C904&yid=A732AF04DDA03BB3&vid=F4B561950EE1D31A&iid=CA4FD0336C81A37A&sid=0401E2DB1F51F8DE&eid=68D88C2FCF9C3098&journal_id=0001-6209&journal_name=微生物学报&referenced_num=3&reference_num=12