Enhancement of pH stability and activity of glycerol dehydratase from Klebsiella pneumoniae by rational design

摘要: Glycerol dehydratase (GDHt) is a key and rate-limiting enzyme in the pathway of 1,3-propanediol (1,3-PD) synthesis. The improvement GDHt’s stability enzymatic activity desirable for biosynthesis 1,3-PD. gldABC gene encoding GDHt Klebsiella pneumoniae was cloned expressed Escherichia coli XL10-Gold, mutation sites were obtained through prediction by PoPMuSiC program. Consequently, two mutants (KpG60 KpG525) developed rational design site-mutagenesis based on 3D structure which constructed from homology modeling. Analyses properties showed that pH about 1.25–2 times higher than wild type, specific activity, Vmax Kcat/Km KpG525 1.5–2 those type. This work presented simple useful measure to improve performance industrial enzyme.