文献类型：期刊文献

英文题名：Development of a Red recombinase system and antisense RNA technology in Klebsiella pneumoniae for the production of chemicals

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

第一作者：李映;Li, Ying

通讯作者：Tian, PF[1]

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

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

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

年份：2016

卷号：6

期号：83

起止页码：79920-79927

外文期刊名：RSC ADVANCES

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000382539600073)】；

基金：This work was supported by grants from National High Technology Research and Development Program (863 Program) (No. 2015AA021003), National Basic Research Program of China (973 Program) (No. 2012CB725200), National Natural Science Foundation of China (No. 21276014, 21476011), Fundamental Research Funds for the Central Universities (YS1407), Innovation Capability Enhancement Program for Universities Governed by Beijing Municipal Commission of Education (No. PXM2015_014209_000010), and Project from Beijing Key Laboratory of Bioactive Substances and Functional Foods in Beijing Union University (ZK70201406).

语种：英文

外文关键词：Acetic acid - Escherichia coli - Genes - Genetic engineering - Glycerol - Lactic acid - Metabolism - pH

摘要：Klebsiella pneumoniae is a promising industrial species, however the lack of genetic engineering tools restricts its applications. Here we developed a lambda (lambda) Red recombinase system and antisense RNA technology in K. pneumoniae to reshape glycerol metabolism pathways. We deleted the lactate dehydrogenase gene ldh through RecA-dependent recombination to block lactic acid synthesis. Next, the 1,3-propanediol dehydrogenase gene dhaT was replaced by an aldehyde dehydrogenase gene (aldH from E. coli) to repress 1,3-propanediol (1,3-PDO) synthesis and simultaneously convert 3-hydroxypropionaldehyde (3-HPA) to 3-hydroxypropionic acid (3-HP). Specially, we developed a Red recombinase system in K. pneumoniae, by which the enzymes related to glycerol metabolism were mutated by transformed oligos. One positive strain produced 6.39 g L-1 3-HP and 32.6 g L-1 1,3-PDO at 36 h without using any antibiotics and inducers. Sequencing results showed that the mutation occurred mainly in byproduct pathways. Finally, antisense RNA technique was applied to block the synthesis of lactic acid and acetic acid. We found that the increase of 3-HP was approximately proportional to the decrease of lactic acid and acetic acid, indicating their competition for glycerol carbon flux. Overall these results and approaches developed in this study provide basis for basic research and microbial production of 3-HP, 1,3-PDO and 2,3-butanediol in K. pneumoniae.