Altering the substrate specificity of glutamate dehydrogenase from Bacillus subtilis by site-directed mutagenesis.
The Lys80, Gly82 and Met101 residues of glutamate dehydrogenase from Bacillus subtilis were mutated into a series of single mutants. The wild-type enzyme was highly specific for 2-oxoglutarate, whereas G82K and M101S dramatically switched to increased specificity for oxaloacetate with kcat values 3.45 and 5.68 s-1, which were 265-fold and 473-fold higher respectively than those for 2-oxoglutarate.
Khan
Iqbal Hassan
IH
Department of Life Science and Biotechnology, Faculty of Life and Environmental Science, Shimane University, Shimane 690-8504, Japan.
Kim
Hyeung
H
Ashida
Hiroyuki
H
Ishikawa
Takahiro
T
Shibata
Hitoshi
H
Sawa
Yoshihiro
Y
D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics; D001665:Binding Sites / Q000502:physiology; D005969:Glutamate Dehydrogenase / Q000737:chemistry* / Q000235:genetics* / Q000502:physiology; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D013379:Substrate Specificity / Q000235:genetics
D005969:Glutamate Dehydrogenase