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Table 1 Effect of external NADH addition on the activity of lyophilized P450 whole-cell catalysts

From: Evaluation of P450 monooxygenase activity in lyophilized recombinant E. coli cells compared to resting cells

Lyophilized E.coli C43 (DE3) harboring Testosterone conversion [%] Formation of 2βhydroxytestosterone [%]
− NADH + NADH − NADH + NADH
pET22b-cyp105D + pCOLADuet-pdx-pdr ≤ 1 3 ≤ 1 3
pET22b-cyp105D + pCOLADuet-pdx-pdr-adh 53 ± 6 72 ± 5 46 ± 4 61± 4
  1. Reaction conditions: 10 mg/mL lyophilized cells in 0.5 mL Phosphate Sucrose EDTA (PSE)-buffer with 1 × nutrient solution, pH 7.5, in 2 mL reaction tubes, 1 mM testosterone 1 dissolved in 5% (v/v) propan-2-ol final concentration, 25 °C, 1100 rpm, 20 h reaction time. 0.25 mM NADH was added up to four times at 0 h, 2 h, 4 and 6 h incubation. For the cells co-expressing the adh, 0.5 mM NADH were added after 4 h. Experiments were performed in technical duplicates