Research Article
Site Saturation Mutagenesis Applications on Candida methylica Formate Dehydrogenase
Table 1
The kinetic parameters of wild-type cmFDH and mutants.
| | Enzymes | NAD+ | NADP+ | (/) NADP+/(/) NAD+ | | (mM) | (s−1) | / | (mM) | (s−1) | / |
| | Native cmFDH | 0.62 ± 0.3 | 0.2 ± 0.1 | 320 | ND | ND | <10−3 | | | 195Ser | 1.1 ± 0.5 | 0.22 ± 0.1 | 220 | 5 ± 1.2 | 0.04 ± 0.09 | 8 | 0.036 | | 197Val | 3.3 ± 0.2 | 0.22 ± 0.1 | 60 | 1.4 ± 1.1 | 0.03 ± 0.05 | 18 | 0.3 | | 195Ser/197Val | 3.9 ± 2.3 | 0.16 ± 0.1 | 40 | 2.2 ± 1.1 | 0.01 ± 0.08 | 4.5 | 0.113 | | 195Ser/197Thr | 0.22 ± 0.03 | 0.2 ± 0.01 | 900 | 4.6 ± 3 | 0.26 ± 0.1 | 56 | 0.06 | | 195Ser/196Leu | 0.7 ± 0.2 | 0.2 ± 0.1 | 260 | 2 ± 0.6 | 0.1 ± 0.02 | 50 | 0.2 | | 195Ser/196Ser | 2.3 ± 1.8 | 0.2 ± 0.05 | 90 | 8.2 ± 4 | 0.16 ± 0.05 | 19 | 0.2 | | 195Ser/196Ala | 0.13 ± 0.1 | 0.2 ± 0.1 | 1460 | 8.5 ± 2.8 | 0.17 ± 0.1 | 20 | 0.014 |
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