Altering the substrate specificity of glutamate dehydrogenase from Bacillus subtilis by site-directed mutagenesis |
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| Authors: | Khan Iqbal Hassan Kim Hyeung Ashida Hiroyuki Ishikawa Takahiro Shibata Hitoshi Sawa Yoshihiro |
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| Affiliation: | Department of Life Science and Biotechnology, Faculty of Life and Environmental Science, Shimane University, Shimane 690-8504, Japan. |
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| Abstract: | 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. |
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