文献类型：期刊文献

英文题名：Enhanced aldehyde dehydrogenase activity by regenerating NAD(+) in Klebsiella pneumoniae and implications for the glycerol dissimilation pathways

作者：Li, Ying[1];Su, Mingyue[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, Coll Biochem Engn, 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：35

期号：10

起止页码：1609-1615

外文期刊名：BIOTECHNOLOGY LETTERS

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000325011900009)】；

基金：This work was supported by National Natural Science Foundation of China (Nos. 20876009, 2107 6013, 21276014) and National Basic Research Program of China (973 Program) (2012CB725200).

语种：英文

外文关键词：Aldehyde dehydrogenase; 3-Hydroxypropionic acid; Klebsiella pneumoniae; NAD(+) regeneration

摘要：In Klebsiella pneumoniae, 3-hydroxypropaldehyde is converted to 3-hydroxypropionic acid (3-HP) by aldehyde dehydrogenase (ALDH) with NAD(+) as a cofactor. Although ALDH overexpression stimulates the formation of 3-HP, it ceases to accumulate when NAD(+) is exhausted. Here we show that NAD(+) regeneration, together with ALDH overexpression, facilitates 3-HP production and benefits cell growth. Three distinct NAD(+)-regenerating enzymes: NADH oxidase and NADH dehydrogenase from K. pneumoniae, and glycerol-3-phosphate dehydrogenase (GPD1) from Saccharomyces cerevisiae, were individually expressed in K. pneumoniae. In vitro assay showed their higher activities than that of the control, indicating their capacities to regenerate NAD(+). When they were respectively co-expressed with ALD4, an ALDH from S. cerevisiae, the activities of ALD4 were significantly elevated compared with that expressing ALD4 alone, suggesting that the regenerated NAD(+) enhanced the activity of ALD4. More interestingly, the growth rates of all NAD(+)-regenerating strains were prolonged in comparison with the control, indicating that NAD(+) regeneration stimulated cell proliferation. This study not only reveals the reliance of ALD4 activity on NAD(+) availability but also provides a method for regulating the dha regulon.