Rates of piceatannol formation by various CYP102A1 mutants
Assays were performed using 100 μM trans-resveratrol as described under Materials and Methods. Values are the mean ± S.D. of triplicate determinations.
Enzyme | Mutated Amino Acid Residues | Piceatannol |
|---|---|---|
| nmol product/min/nmol P450 | ||
| Wild-type | 0.22 ± 0.01 | |
| Mutant | ||
| #1 | F87A | 0.021 ± 0.001 |
| #2 | A264G | N.D. |
| #3 | F87A/A264G | N.D. |
| #4 | R47L/Y51F | 0.027 ± 0.001 |
| #5 | R47L/Y51F/A264G | N.D. |
| #6 | R47L/Y51F/F87A | N.D. |
| #7 | R47L/Y51F/F87A/A264G | N.D. |
| #8 | A74G/F87V/L188Q | 1.1 ± 0.1 |
| #9 | R47L/L86I/L188Q | 0.24 ± 0.01 |
| #10 | R47L/F87V/L188Q | 1.6 ± 0.1 |
| #11 | R47L/F87V/L188Q/E267V | 0.64 ± 0.03 |
| #12 | R47L/L86I/L188Q/E267V | 0.33 ± 0.03 |
| #13 | R47L/L86I/F87V/L188Q | 4.0 ± 0.1 |
| #14 | R47L/F87V/E143G/L188Q/E267V | 1.3 ± 0.1 |
| #15 | R47L/E64G/F87V/E143G/L188Q/E267V | 1.9 ± 0.3 |
| #16 | R47L/F81I/F87V/E143G/L188Q/E267V | 0.97 ± 0.1 |
| #17 | R47L/E64G/F81I/F87V/E143G/L188Q/E267V | 1.8 ± 0.1 |
N.D., not detectable (the rate of product formation was less than 0.001 nmol product/min/nmol P450).