title: Saturation-mutagenesis in two positions distant from active site of a Klebsiella pneumoniae glycerol dehydratase identifies some highly active mutants.
abstract: Synthesis of 1,3-propanediol (1,3-PD) from glycerol through the biotransformation process requires two steps, catalyzed by glycerol dehydratase (GDHt) and 1,3-PD oxidoreductase. GDHt is the rate-limiting enzyme in this process. All recombinant microorganisms for production of 1,3-PD so far utilized the natural genes that may not have been optimized. Two positions, which are 19.3A and 29.6A away from the active site in GDHt from Klebsiella pneumoniae, were subjected to saturation-mutagenesis and 38 mutants were characterized. The catalytic activity of a mutant in beta-subunit (beta-Q42F, 29.6A from the active site) was 8.3-fold higher than the wild type, and the enzyme efficiency of other two mutants beta-Q42L and beta-Q42S for substrate glycerol was 336-fold and 80-fold higher than that for 1,2-propanediol. This investigation supplied further evidence that distant mutations could be a good source of diversity and therefore, made a contribution to the toolbox of industrial enzyme improvement.
authors: Guangxi Academy of Sciences, Nanning, China.
authors: Xianghui
authors: X
authors: Qi
authors: Yunlai
authors: Y
authors: Chen
authors: Ke
authors: K
authors: Jiang
authors: Wenpu
authors: W
authors: Zuo
authors: Zhaofei
authors: Z
authors: Luo
authors: Yutuo
authors: Y
authors: Wei
authors: Liqin
authors: L
authors: Du
authors: Hang
authors: H
authors: Wei
authors: Ribo
authors: R
authors: Huang
authors: Qishi
authors: Q
authors: Du
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry; D020134:Catalytic Domain; D004795:Enzyme Stability; D057166:High-Throughput Screening Assays; D006836:Hydro-Lyases / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification; D006863:Hydrogen-Ion Concentration; D007711:Klebsiella pneumoniae / Q000201:enzymology*; D008958:Models, Molecular; D016296:Mutagenesis / Q000235:genetics*; D050505:Mutant Proteins / Q000737:chemistry; D009154:Mutation / Q000235:genetics*; D017433:Protein Structure, Secondary; D013329:Structure-Activity Relationship; D013696:Temperature
chemical_list: D001426:Bacterial Proteins; D050505:Mutant Proteins; D006836:Hydro-Lyases; C020671:glycerol dehydratase