Novel iron-sulfur containing NADPH-Reductase from Nocardia farcinica IFM10152 and fusion construction with CYP51 lanosterol demethylase

Title
Novel iron-sulfur containing NADPH-Reductase from Nocardia farcinica IFM10152 and fusion construction with CYP51 lanosterol demethylase
Author(s)
최권영[최권영]정은옥[정은옥]정다혜[정다혜]비쉬누 판데이[비쉬누 판데이]이나훔[이나훔]윤형돈박형연[박형연]김병기[김병기]
Keywords
SUFFICIENT P450 MONOOXYGENASE; 14-ALPHA-DEMETHYLASE CYP51; CYTOCHROME-P450 ENZYMES; EXPRESSION; CLONING; FLAVOCYTOCHROME; EVOLUTIONARY
Issue Date
201203
Publisher
WILEY-BLACKWELL
Citation
BIOTECHNOLOGY AND BIOENGINEERING, v.109, no.3, pp.630 - 636
Abstract
CYP51, a sterol 14a-demethylase, is one of the key enzymes involved in sterol biosynthesis and requires electrons transferred from its redox partners. A unique CYP51 from Nocardia farcinica IFM10152 forms a distinct cluster with ironsulfur containing NADPH-P450 reductase (FprD) downstream of CYP51. Previously, sequence alignment of nine reductases from N. farcinica revealed that FprC, FprD, and FprH have an additional sequence at their N-termini that has very high identity with ironsulfur clustered ferredoxin G (FdxG). To construct an artificial self-sufficient cytochrome P450 monooxygenase (CYP) with only FprD, CYP51, and ironsulfur containing FprD were fused together with designed linker sequences. CYP51FprD fusion enzymes showed distinct spectral properties of both flavoprotein and CYP. CYP51FprD F1 and F2 in recombinant Escherichia coli BL21(DE3) catalyzed demethylation of lanosterol more efficiently, with kcat/Km values of 96.91 and 105.79 nmol/min/nmol, respectively, which are about 35-fold higher compared to those of CYP51 and FprD alone. Biotechnol. Bioeng. 2012; 109:630636. (C) 2011 Wiley Periodicals, Inc.
URI
http://hdl.handle.net/YU.REPOSITORY/29563http://dx.doi.org/10.1002/bit.24359
ISSN
0006-3592
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생명공학부 > 생명공학부 > Articles
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