TY - JOUR
T1 - Recombinant glycerol dehydratase from Klebsiella pneumonia XJPD-Li
T2 - Induction optimization, purification and characterization
AU - Xu, X. L.
AU - Zhang, G. L.
AU - Lv, B.
AU - Yuan, Y. J.
AU - Li, C.
PY - 2011/3
Y1 - 2011/3
N2 - 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 Km 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.
AB - 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 Km 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.
UR - http://www.scopus.com/inward/record.url?scp=79952414746&partnerID=8YFLogxK
U2 - 10.1134/S0003683811020189
DO - 10.1134/S0003683811020189
M3 - Article
AN - SCOPUS:79952414746
SN - 0003-6838
VL - 47
SP - 144
EP - 150
JO - Applied Biochemistry and Microbiology
JF - Applied Biochemistry and Microbiology
IS - 2
ER -