cholecalciferol and mevastatin

Topics: Actinobacteria; Amino Acid Substitution; Bacterial Proteins; Cholecalciferol; Conserved Sequence; Cytochrome P-450 Enzyme System; Ferredoxins; Gene Expression; Hydroxylation; Isoenzymes; Lovastatin; Molecular Docking Simulation; Mutation; Pravastatin; Protein Engineering; Streptomyces; Substrate Specificity

We report here some efficient biotransformations using Escherichia coli strains with disruptions for the AcrAB-TolC efflux pump system. Biotransformations of compactin into pravastatin (6alpha-hydroxy-iso-compactin) were performed using E. coli strains with tolC and/or acrAB mutations expressing a cytochrome P450 (P450) gene. The production levels of pravastatin using strains with acrAB, tolC, and tolC acrAB mutations increased by 3.7-, 7.0-, and 7.1-fold, respectively. Likewise, the production levels of 25-hydroxy vitamin D3 and 25-hydroxy 4-cholesten 3-one using tolC acrAB mutant strains expressing an individual P450 gene increased by 2.2- and 16-fold, respectively. The enhancement of this biotransformation efficiency could be explained by increases in the intracellular amounts of substrates and the concentrations of active P450s. These results demonstrate that we have achieved versatile methods for efficient biotransformations using E. coli strains with tolC acrAB mutations expressing P450 genes.

Topics: Bacterial Outer Membrane Proteins; Calcifediol; Cholecalciferol; Cholestenones; Cytochrome P-450 Enzyme System; Escherichia coli; Escherichia coli Proteins; Hydrophobic and Hydrophilic Interactions; Intracellular Space; Lovastatin; Membrane Transport Proteins; Mutation; Oxidation-Reduction; Pravastatin; Spectrum Analysis; Stereoisomerism; Substrate Specificity