|
KMID : 0545120130230121699
|
|
Journal of Microbiology and Biotechnology
2013 Volume.23 No. 12 p.1699 ~ p.1707
|
|
|
Characterization and Cofactor Binding Mechanism of a Novel NAD(P)H-Dependent Aldehyde Reductase from Klebsiella pneumoniae DSM2026
|
|
Cheng-Wei Ma
Le Zhang
Jian-Ying Dai
Zhi-Long Xiu
|
|
|
Abstract
|
|
|
|
During the fermentative production of 1,3-propanediol under high substrate concentrations, accumulation of intracellular 3-hydroxypropionaldehyde will cause premature cessation of cell growth and glycerol consumption. Discovery of oxidoreductases that can convert 3- hydroxypropionaldehyde to 1,3-propanediol using NADPH as cofactor could serve as a solution to this problem. In this paper, the yqhD gene from Klebsiella pneumoniae DSM2026, which was found encoding an aldehyde reductase (KpAR), was cloned and characterized. KpAR showed broad substrate specificity under physiological direction, whereas no catalytic activity was detected in the oxidation direction, and both NADPH and NADH can be utilized as cofactors. The cofactor binding mechanism was then investigated employing homology modeling and molecular dynamics simulations. Hydrogen-bond analysis showed that the hydrogen-bond interactions between KpAR and NADPH are much stronger than that for NADH. Free-energy decomposition dedicated that residues Gly37 to Val41 contribute most to the cofactor preference through polar interactions. In conclusion, this work provides a novel aldehyde reductase that has potential applications in the development of novel genetically engineered strains in the 1,3-propanediol industry, and gives a better understanding of the mechanisms involved in cofactor binding.
|
|
|
KEYWORD
|
|
|
1,3-Propanediol, Aldehyde reductase, Cofactor binding, Free energy decomposition, Klebsiella pneumoniae
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
  
|
|