文献类型：期刊文献

英文题名：Gene Arrangements in Expression Vector Affect 3-Hydroxypropionic Acid Production in Klebsiella pneumoniae

作者：Li, Ying[1];Ge, Xizhen[2];Tian, Pingfang[1]

第一作者：Li, Ying

通讯作者：Tian, PF[1]

机构：[1]Beijing Univ Chem Technol, Coll Life Sci & Technol, Beijing 100029, Peoples R China;[2]Beijing Union Univ, Biochem Engn Coll, Beijing 100023, Peoples R China

第一机构：Beijing Univ Chem Technol, Coll Life Sci & Technol, Beijing 100029, Peoples R China

通讯机构：[1]corresponding author), Beijing Univ Chem Technol, Coll Life Sci & Technol, Beijing 100029, Peoples R China.

年份：2013

卷号：53

期号：4

起止页码：418-424

外文期刊名：INDIAN JOURNAL OF MICROBIOLOGY

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000324646900007)】；

基金：This work was supported by National Natural Science Foundation of China (No. 20876009, 21076013, 21276014) and National Basic Research Program of China (973 Program) (2012CB725200).

语种：英文

外文关键词：Klebsiella pneumoniae; 3-Hydroxypropionic acid; Gene arrangement; Glycerol dehydratase; Aldehyde dehydrogenase

摘要：Biosynthesis of 3-hydroxypropionic acid (3-HP) typically involves two sequential reactions catalyzed by glycerol dehydratase (DhaB) and aldehyde dehydrogenase (AldH). Although plasmid-dependent over-expression of the two enzymes is common, systematic investigation of gene arrangement in vector has not been reported. Here we show that gene arrangements have a noticeable influence on 3-HP production. Using Klebsiella pneumoniae as a host, three AldH-coding genes: ald4 from Saccharomyces cerevisiae, aldh from Escherichia coli, and puuC from host K. pneumoniae, were respectively ligated to dhaB. The recombinant Kp/pET-pk-ald4-dhaB (Kp refers to as K. pneumoniae, pk is a native promoter) produced the highest yield of 3-HP in comparison to both Kp/pET-pk-dhaB-ald4 and Kp/pET-pk-dhaB-pk-ald4, suggesting that the preferential expression of AldH can increase 3-HP production. Additionally, when different AldH-coding genes were respectively ligated downstream of dhaB, the recombinant Kp/pET-pk-dhaB-puuC produced more 3-HP than that by Kp/pET-pk-dhaB-aldh or Kp/pET-pk-dhaB-ald4, implying the intrinsic compatibility of native gene puuC with its host. These findings indicate the applicability of native AldH-coding gene and provide insights into strategies for metabolic engineering of multiple genes.