Table 1 GC–MS analysis of the enzyme assay products.
Amino acid | Putative function | Mutant | Sodorifen | pre-sodorifen production |
|---|---|---|---|---|
Amino acids of the active site | ||||
Y39 | Catalytic base, catalytic dyad | Y39A | No | No |
E42 | Mg2+ coordination | E42A | No | No |
H45 | Pyrophosphate stability | H45A | 25% ± 18% | 25% ± 11% |
W46 | Carbocation stabilization, hydrophobic pocket | W46A | No | No |
F56 | Carbocation stabilization, hydrophobic pocket | F56A | No | No |
Y61 | Correct position of SAM in the active site | Y61A | No | No |
H191 | Catalytic base, catalytic dyad | H191A | No | No |
N219 | Closes active site and helps correct positioning of FPP | N219A | No | No |
D237 | Enzyme stability | D237A | No | No |
L239 | Hydrophobic pocket, enzyme stability | L239A | 20% ± 10% | 23% ± 14% |
C241 | Enzyme stability | C241A | 20% ± 8% | 22% ± 15% |
D272 | Enzyme stability | D272A | No | No |
E297 | Stabilizes tertiary structure for correct recognition of pyrophosphate | E297A | 25% ± 17% | 27% ± 7 |
W306 | Carbocation stabilization, hydrophobic pocket | W306A | No | No |
Wild Type | 100% ± 8% | 100% ± 3% | ||
