TABLE 3.
Product formation for bioreduction of ethyl 3-keto-4,4,4-trifluorobutyrate 1 with coupled permeabilized cells
| B. pumilus Phe-C3 concn (g [dry wt] of cells/liter) | E. coli BL21 concn (g [dry wt] of cells/liter)a | Ethyl 3-keto-4,4,4-triflurobutyrate concn (mM) | Glucose concn (mM) | Activity (U/g [dry wt] of cells)b | Time (h) | NADP+ concn (mM) | (R)-Ethyl 3-hydroxy-4,4,4-triflurobutyrate concn (mM) | % Conversion | TTN |
|---|---|---|---|---|---|---|---|---|---|
| 40 | 20 | 60 | 440 | 1.3 | 0 | 0.005 | |||
| 20 | +0.005c | 19.0 | 32 | 3,800 | |||||
| 44 | +0.005 | 36.0 | 60 | 3,600 | |||||
| 68 | 50.5 | 84 | 3,400 | ||||||
| 40 | 20 | 60 + 60d | 440 | 1.3 | 0 | 0.005 | |||
| 24 | +0.005 | 20.0 | 17 | 4,000 | |||||
| 48 | +0.005 | 38.0 | 31 | 3,800 | |||||
| 72 | +0.005 | 52.5 | 44 | 3,500 | |||||
| 96 | 64.0 | 53 | 3,200 |
a
Permeabilized E. coli BL21(pGDH1) cells with an activity of 61 U/g (dry weight) of cells were used.
b
Activity was determined over the first 1 h.
c
+0.005 indicates that 0.005 mM NADP+ was added at the time indicated.
d
Ethyl 3-keto-4,4,4-trifluorobutyrate 1 was added at the beginning (60 mM) and at 5 h (60 mM).