Recombinant glycerol dehydratase from <Emphasis Type="Italic">Klebsiella pneumonia</Emphasis> XJPD-Li: induction optimization,purification and characterization |
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| Authors: | X L Xu G L Zhang B Lv Y -J Yuan C Li |
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| Institution: | 1.School of Chemical Engineering and Technology,Tianjin University,Tianjin,P.R. China;2.School of Life Science and Technology,Beijing Institute of Technology,Beijing,P.R. China;3.Key Laboratory for Green Processing of Chemical Engineering of Xinjiang Bingtuan,Shihezi University,Xinjiang,P. R. China |
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| Abstract: | Glycerol dehydratase (GDHt) is the rate limiting enzyme in the biosynthesis of 1,3-propanediol from glycerol. The optimization
of inducting process for recombinant GDHt from Klebsiella pneumoniae XJPD-Li carried out to increase specific activity and ratio of soluble form. The optimum condition was inducing under the
isopropyl-β-D-thiogalactoside concentration of 0.8 mM and the temperature of 20°C for 3 h. Homogeneity of GDHt then was obtained
by affinity chromatography, resulted in 2.11-fold purification and an overall yield of 47.5%. The optimum pH and reaction
temperature of GDHt were pH 8.0 and 45°C, respectively. The K
m for glycerol, 1,2-propanediol, 1,2-ethanediol and coenzyme B12 were 0.48 mM, 1.43 mM, 3.07 mM, and 10.03 nM, respectively.
The GDHt showed relatively stable even under temperature of 40°C and a bit blunt to oxygen. The thermo-inactivation kinetic
models were fit linear under different temperatures. |
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