Table 1. Cofactor specificities of the WT and mutant FDH proteins.
| NADP+ reduction (µM/h-1) | NAD+ reduction (µM/h-1) | NADP+/NAD+ | |
|---|---|---|---|
| Pseudomonas | |||
| WT (DH) | 24.0 ± 1.7 | 1010 ± 44 | 0.024 |
| EN | 30.0 ± 0.5 | 417 ± 45 | 0.072 |
| SN | 279 ± 1 | 579 ± 30 | 0.48 |
| SH | 184 ± 2 | 429 ± 10 | 0.43 |
| AN | 141 ± 6 | 138 ± 2 | 1.0 |
| AH | 153 ± 2 | 203 ± 10 | 0.75 |
| QN | 293 ± 5 | 30.9 ± 4.2 | 9.5 |
| QH | 299 ± 13 | 31.0 ± 3.9 | 9.6 |
| Candida | |||
| WT (DYH) | 16.8 ± 0.6 | 302 ± 4 | 0.056 |
| QRN | 265 ± 0.3 | 51.0 ± 9.6 | 5.2 |
| Thiobacillus | |||
| WT (DH) | 18.9 ± 0.4 | 926 ± 6 | 0.020 |
| SN | 0.4 ± 0.2 | 3.9 ± 2.9 | 0.10 |
| SH | 3.0 ± 1.2 | 2.6 ± 1.2 | 1.15 |
| AN | 0 ± 0 | 3.1 ± 2.6 | 0 |
| AH | 0.2 ± 0.1 | 3.2 ± 2.6 | 0.063 |
| QN | 0.8 ± 0.1 | 0.3 ± 0.1 | 2.6 |
| QH | 0.6 ± 0.4 | 0.4 ± 0.2 | 1.5 |
| Arabidopsis | |||
| WT (DL) | 1.3 ± 0.2 | 416 ± 61 | 0.0031 |
| SN | 6.8 ± 3.0 | 5.4 ± 4.0 | 1.3 |
| SH | 4.5 ± 2.6 | 4.9 ± 4.4 | 0.92 |
| AN | 5.8 ± 3.0 | 3.1 ± 2.3 | 1.9 |
| AH | 14.6 ± 1.7 | 3.3 ± 0.9 | 4.4 |
| QN | 3.4 ± 2.4 | 5.1 ± 1.6 | 0.67 |
| QH | 70.5 ± 13.9 | 4.2 ± 2.7 | 17 |
| Potato | |||
| WT (DL) | 6.7 ± 0.2 | 803 ± 3 | 0.0083 |
| EN | 5.5 ± 1.5 | 112 ± 1 | 0.049 |
| SN | 53.1 ± 0.7 | 8.6 ± 3.1 | 6.2 |
| SH | 91.7 ± 5.5 | 75.5 ± 1.8 | 1.2 |
| AH | 116 ± 2 | 11.1 ± 3.8 | 10 |
| QN | 93.4 ± 1.4 | 3.7 ± 2.3 | 25.2 |
| QH | 149 ± 11 | 3.4 ± 1.0 | 44 |
FDHs were added up to a final concentration of 5 µg/mL into PBS containing 0.8 mM NAD+ or NADP+, 50 mM sodium formate, and 0.05% Triton X-100 at 30° C. NADH or NADPH production was monitored at 340 nm. All the measurements were performed in triplicate. The two-letter or three-letter descriptions in the left column indicate 2 or 3 one-letter codes of amino acid residue at the position of 222 and 224 in Pseudomonas , 195, 196 and 197 in Candida , 197 and 199 in potato, 227 and 229 in Arabidopsis , and 222 and 224 in Thiobacillus .