Identification of a new plasmid-encoded cytochrome P450 CYP107DY1 from Bacillus megaterium with a catalytic activity towards mevastatin.
In the current work, we describe the identification and characterization of the first plasmid-encoded P450 (CYP107DY1) from a Bacillus species. The recombinant CYP107DY1 exhibits characteristic P450 absolute and reduced CO-bound difference spectra. Reconstitution with different redox systems revealed the autologous one, consisting of BmCPR and Fdx2, as the most effective one. Screening of a library of 18 pharmaceutically relevant compounds displayed activity towards mevastatin to produce pravastatin. Pravastatin is an important therapeutic drug to treat hypercholesterolemia, which was described to be produced by oxyfunctionlization of mevastatin (compactin) by members of CYP105 family. The hydroxylation at C6 of mevastatin was also suggested by docking this compound into a computer model created for CYP107DY1. Moreover, in view of the biotechnological application, CYP107DY1 as well as its redox partners (BmCPR and Fdx2) were successfully utilized to establish an E. coli based whole-cell system for an efficient biotransformation of mevastatin. The in vitro and in vivo application of the CYP07DY1 also offers the possibility for the screening of more substrates, which could open up further biotechnological usage of this enzyme.
Institute of Biochemistry, Saarland University, 66123 Saarbrücken, Germany.
Mohammed
M
Milhim
Institute of Biochemistry, Saarland University, 66123 Saarbrücken, Germany.
Natalia
N
Putkaradze
Institute of Biochemistry, Saarland University, 66123 Saarbrücken, Germany.
Ammar
A
Abdulmughni
Institute of Biochemistry, Saarland University, 66123 Saarbrücken, Germany.
Fredy
F
Kern
Institute of Biochemistry, Saarland University, 66123 Saarbrücken, Germany.
Philip
P
Hartz
Institute of Biochemistry, Saarland University, 66123 Saarbrücken, Germany. Electronic address: ritabern@mx.uni-saarland.de.
Rita
R
Bernhardt
D001410:Bacillus megaterium / Q000378:metabolism*; D001711:Biotransformation; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D004926:Escherichia coli; D006900:Hydroxylation; D008148:Lovastatin / Q000031:analogs & derivatives / Q000378:metabolism; D010957:Plasmids; D017035:Pravastatin / Q000378:metabolism; D013379:Substrate Specificity
C012258:mevastatin; D003577:Cytochrome P-450 Enzyme System; D008148:Lovastatin; D017035:Pravastatin
Bacillus megaterium; Bacterial cytochrome P450; BmCPR; CYP107 family; Fdx2; Mevastatin; Pravastatin