Biotransformation of flavone by CYP105P2 from Streptomyces peucetius.
Biocatalytic transfer of oxygen in isolated cytochrome P450 or whole microbial cells is an elegant and efficient way to achieve selective hydroxylation. Cytochrome P450 CYP105P2 was isolated from Streptomyces peucetius that showed a high degree of amino acid identity with hydroxylases. Previously performed homology modeling, and subsequent docking of the model with flavone, displayed a reasonable docked structure. Therefore, in this study, in a pursuit to hydroxylate the flavone ring, CYP105P2 was co-expressed in a two-vector system with putidaredoxin reductase (camA) and putidaredoxin (camB) from Pseudomonas putida for efficient electron transport. HPLC analysis of the isolated product, together with LCMS analysis, showed a monohydroxylated flavone, which was further established by subsequent ESI/MS-MS. A successful 10.35% yield was achieved with the whole-cell bioconversion reaction in Escherichia coli. We verified that CYP105P2 is a potential bacterial hydroxylase.
Institute of Biomolecule Reconstruction (iBR), Department of Pharmaceutical Engineering, SunMoon University, Asansi 336-708, Korea.
Narayan Prasad
NP
Niraula
Saurabh
S
Bhattarai
Na-Rae
NR
Lee
Jae Kyung
JK
Sohng
Tae-Jin
TJ
Oh
D001711:Biotransformation; D002851:Chromatography, High Pressure Liquid; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005288:Ferredoxins / Q000235:genetics / Q000378:metabolism; D047309:Flavones / Q000378:metabolism*; D006900:Hydroxylation; D013058:Mass Spectrometry; D060847:Metabolic Engineering; D009247:NADH, NADPH Oxidoreductases / Q000235:genetics / Q000378:metabolism; D016958:Pseudomonas putida / Q000201:enzymology; D013302:Streptomyces / Q000201:enzymology* / Q000235:genetics
D005288:Ferredoxins; D047309:Flavones; C100137:putidaredoxin; D003577:Cytochrome P-450 Enzyme System; D009247:NADH, NADPH Oxidoreductases; C022486:putidaredoxin reductase; C043562:flavone