Efficient biotransformations using Escherichia coli with tolC acrAB mutations expressing cytochrome P450 genes.
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.
Bioresource Laboratories, Mercian Corporation, 1808 Nakaizumi, Iwata, Shizuoka 438-0078, Japan. fujii-td@mercian.co.jp
Tadashi
T
Fujii
Yoshikazu
Y
Fujii
Kazuhiro
K
Machida
Atsushi
A
Ochiai
Masashi
M
Ito
D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D002112:Calcifediol / Q000378:metabolism; D002762:Cholecalciferol / Q000378:metabolism; D002783:Cholestenones / Q000737:chemistry / Q000378:metabolism; D003577:Cytochrome P-450 Enzyme System / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D004926:Escherichia coli / Q000166:cytology / Q000235:genetics* / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics*; D057927:Hydrophobic and Hydrophilic Interactions; D042541:Intracellular Space / Q000378:metabolism; D008148:Lovastatin / Q000031:analogs & derivatives / Q000378:metabolism; D026901:Membrane Transport Proteins / Q000235:genetics*; D009154:Mutation; D010084:Oxidation-Reduction; D017035:Pravastatin / Q000378:metabolism; D013057:Spectrum Analysis; D013237:Stereoisomerism; D013379:Substrate Specificity
D001425:Bacterial Outer Membrane Proteins; D002783:Cholestenones; D029968:Escherichia coli Proteins; D026901:Membrane Transport Proteins; C073126:tolC protein, E coli; D002762:Cholecalciferol; C012258:mevastatin; C000475:cholest-4-en-3-one; D003577:Cytochrome P-450 Enzyme System; D008148:Lovastatin; D017035:Pravastatin; D002112:Calcifediol