%0 Journal Article
%T Construction of recombinant Saccharomyces cerevisiae producing 1,3-propanediol by one step method<br>一步法产1,3-丙二醇酿酒酵母基因工程菌的构建
%A MA Zheng
%A RAO Zhi-ming
%A SHEN Wei
%A FANG Hui-ying
%A ZHUGE Jian
%A <br>马正
%A 饶志明
%A 沈微
%A 方慧英
%A 诸葛健
%J 微生物学报
%D 2007
%I 
%X 1,3-Propanediol is one of the most important industrial chemicals for its highly desired properties and its wide applications as a key component of an emerging polymer business. Biological production of 1,3-propanediol has been a novel and competitive way. In our previous job, the gene dahB encoding for glycerol dehydratase from Klebsiella and the gene yqhD encoding for 1,3-propanediol oxidoreductase isoenzyme from E. coli were cloned respectively. The two genes were then tandemly ligated and expressed successfully in E. coli. The recombinant E. coli strain could produce 1,3-propanediol from D-glycerol. In the current research, the expression vectors including pGAPZB-yqhD, pGAPZB-dhaB and pYX212-zeocin-pGAP-yqhD-pGAP-dhaB were furtherly constructed on the basis of our previous job. Then the vector pYX212-zeocin-pGAP-yqhD-pGAP-dhaB was introduced into Saccharomyces cerevisiae W303-1A using LiAc transformation method successfully. D-glucose is used as substrate to produce 1,3-propanediol with the recombinant strain after fermentation for 72h. SDS-PAGE analysis showed recombinant products at about 61kD, 43kD, 21kD, 15kD, consistent with the molecular weight from the report. The specific enzymatic activity of the glycerol dehydratase and 1,3-propanediol oxidoreductase isoenzyme of the recombinant yeast strain S. cerevisiae W303-1A/pYX2l2-zeocin-pGAP-yqhD-pGAP-dhaB were 24U/mg protein and 15U/mg protein, respectively, while those of the control were undetectable. In contrast to the wild strain without 1,3-propanediol output, 1,3-propanediol concentration of the recombinant yeast strain S. cerevisiae W303-1A/pYX212-zeocin-pGAP-yqhD-pGAP-dhaB reaches about 1.5g/L. The above results showed that the engineered S. cerevisiae strain which can convert the D-glucose as substrate to produce 1,3-propanediol by one-step fermentation was constructed successfully. This accomplishment bodes well for future construction of recombinant yeast strain which could overproduce 1,3-propanediol with the lower cost feedstock D-glucose by introducing the two genes yqhD and dhaB into the yeast strain overproducing glycerol with D-glucose (e.g. Candida glycerinogenes WL2002-5, which is capable of producing glycerol more than 120g/L with D-glucose as substrate and has been used for the commercial production of glycerol).
%K one-step fermentation
%K 1
%K 3-Propanediol
%K engineered yeast strain<br>一步法发酵
%K 1
%K 3-丙二醇
%K 酿酒酵母工程菌
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A3C6BA55AB623B90FA9104CFFC826F3C&aid=0F2FD9BF00FE651F&yid=A732AF04DDA03BB3&vid=F4B561950EE1D31A&iid=E158A972A605785F&sid=4720E9D07E8A2290&eid=70E3F4DEB0172F14&journal_id=0001-6209&journal_name=微生物学报&referenced_num=0&reference_num=22