Table 3.
Reduction of activated alkenes by PETNR over 24–72 h hours with NADPH and an NADP+/G6PDH co-factor regeneration system, respectively.[a]
| Entry | Conf. | Product | Cofactor | 24 h Conv.[b] [%] |
Yield[b] [%] | ee[c] [%] | 48 h ee[c] [%] |
72 h ee[c] [%] |
|---|---|---|---|---|---|---|---|---|
| 1 |
|
NADPH | >99 | >99 | 93 | - | - | |
| 2 | (R)-1b | NADP+/G6PDH | >99 | >99 | 99 | >99 | >99 | |
| 3 |
|
NADPH | >99 | >99 | 21 | - | - | |
| 4 | (R)-3b | NADP+/G6PDH | >99 | >99 | 57 | 26 | 13 | |
| 5 |
|
NADPH | >99 | >99 | 94 | - | - | |
| 6 | (R)-5b | NADP+/G6PDH | >99 | >99 | 96 | 94 | 91 | |
| 7 |
|
NADPH | >99 | 88 | 93% de[b] | - | - | |
| 8 | (2R,5R)-6b | NADP+/G6PDH | >99 | 81 | 94% de[b] | 92% de[b] | 89% de[b] | |
| 9 |
|
NADPH | >99 | 97 | 81% deb | — | — | |
| 10 | (2R,5S)-7b | NADP+/G6PDH | >99 | 91 | 87% de[b] | 80% de[b] | 75% de[b] | |
| 11 |
|
NADPH | 54 | 53 | >99 | - | - | |
| 12 | (S)-15b | NADP+/G6PDH | 53 | 53 | >99 | >99 | >99 | |
| 13 | (S)-16b |
|
NADPH | 44 | 41 | >99 | - | - |
| 14 | NADP+/G6PDH | 41 | 40 | >99 | >99 | >99 | ||
| 15 |
|
NADPH | >99 | >99 | 65 | - | - | |
| 16 | (S)-17b | NADP+/G6PDH | >99 | >99 | 71 | 65 | 58 |
[a]
Conditions for assays with NADPH: alkene (5 mM), PETN reductase (2 μM), NADPH (15 mM) in phosphate buffer (50 mM, 1 mL total volume), 30 °C at 130 rpm, 24 h; Conditions for assays with NADP+/G6PDH: alkene (5 mM), PETN reductase (2 μM), glucose 6-phosphate dehydrogenase (10 U), glucose 6-phosphate (15 mM) and NADP+ (10 μM) in phosphate buffer (50 mM, 1 mL total volume), 30 °C at 130 rpm, 24 h. Alkenes 1a and 3a were added as a DMF solution (2% final DMF solution).
[b]
By GC using a DB-Wax column.
[c]
By GC or HPLC. Conv. = % conversion.