Improvement of P450BM-3 whole-cell biocatalysis by integrating heterologous cofactor regeneration combining glucose facilitator and dehydrogenase in E. coli

摘要: Escherichia coli BL21, expressing a quintuple mutant of P450BM-3, oxyfunctionalizes α-pinene in an NADPH-dependent reaction to oxide, verbenol, and myrtenol. We optimized the whole-cell biocatalyst by integrating recombinant intracellular NADPH regeneration system through co-expression glucose facilitator from Zymomonas mobilis for uptake unphosphorylated NADP+-dependent dehydrogenase Bacillus megaterium that oxidizes gluconolactone. The engineered strain showed nine times higher initial oxide formation rate corresponding sixfold yield 20 mg g−1 cell dry weight after 1.5 h. total product was 1,000 μmol h−1 μmol−1 P450 leading 32 oxidized products per gram physiological functioning heterologous cofactor illustrated sevenfold increased presence compared glucose-free conditions.