Expression of Synthetic cyp102A1-LG23 Gene and Functional Analysis of Recombinant Cytochrome P450 BM3-LG23 in the Actinobacterium Mycolicibacterium smegmatis

In this study, syntheticcyp102A1-LG23 gene encoding the P450 BM3 mutant was expressed as a component of either monocistronic operon or bicistronic operon containing thegdh (glucose dehydrogenase, GDH) orzwf2 (glucose 6-phosphate dehydrogenase, G6PD) gene inMycolicibacterium smegmatis BD cells. The recombinant bacteria were able hydroxylate androst-4-ene-3,17-dione (AD) into 7 β-OH-AD. Their biocatalytic activity was increased twice by increasing the solubility of CYP102A1-LG23 protein in the cells and supplementing the cells with the additional cofactor regeneration system by introducing GDH and G6PD. The maximum 7β-OH-AD yield (37.68 mol%) was achieved by co-expressi on ofcyp102A1-LG23 andgdh genes inM.  smegmatis. These results demonstrate the possibility of using synthetic genes to obtain recombinant enzymes and expand our understanding of the processes involved in steroid hydroxylation by bacterial cytochromes. The data obtained can be used to develop new approaches for microbiological production of 7 β-hydroxylated steroids in genetically modifiedMycolicibacterium species.
Source: Biochemistry (Moscow) - Category: Biochemistry Source Type: research