含甘油脱水酶激活因子编码基因的产1,3-丙二醇新型重组菌的构建

Construction of Novel Recombinant Strain Harboring Glycerol Dehydratase Reactivating Factor Capable of Producing 1,3-propanediol

利用PCR技术扩增来源于弗氏柠檬杆菌(Citrobacter freundii)的甘油脱水酶编码基因dhaB以及甘油脱水酶激活因子编码基因dhaGdhaF,将其与1,3-丙二醇氧化还原酶同工酶的编码基因yqhD串联在温控表达载体pHsh上,构建重组菌E.coliJM109(pHsh-dhaB-dhaG-dhaF-yqhD)。SDS-PAGE分析显示,融合表达产物的分子量同核酸序列测定的推导值相符。与未串联甘油脱水酶激活因子编码基因的重组菌E.coliJM109(pHsh-dhaB-yqhD)相比,1,3-丙二醇的产量提高了28%。

The dhaB gene encoding glycerol dehydratase and dhaG dhaF gene encoding glycerol dehydratase reactivating factor from Citrobacter freundii were amplified by PCR.The temperature control expression vector pHsh harboring yqhD,dhaB,dhaG and dhaF gene was transformed into E.coli JM109 to yield the recombinant strain E.coli JM109（pHsh-dhaB-dhaG-dhaF-yqhD）.The results from SDS-PAGE analysis show that the recombinant product was consistent with the molecular weight predicted from gene sequence.The fermentation result show that the yield of 1,3-propanediol was increased by 28% compared with E.coli JM109（pHsh-dhaB-yqhD）.