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Sequence-based screening for self-sufficient P450 monooxygenase from a metagenome library

Title
Sequence-based screening for self-sufficient P450 monooxygenase from a metagenome library
Authors
Kim B.S.Kim S.Y.Park J.Park W.Hwang K.Y.Yoon Y.J.Oh W.K.Kim B.Y.Ahn J.S.
Ewha Authors
윤여준
SCOPUS Author ID
윤여준scopus
Issue Date
2007
Journal Title
Journal of Applied Microbiology
ISSN
1364-5072JCR Link
Citation
vol. 102, no. 5, pp. 1392 - 1400
Indexed
SCI; SCIE; SCOPUS WOS scopus
Abstract
Aims: Cytochrome P450 monooxygenases (CYPs) are useful catalysts for oxidation reactions. Self-sufficient CYPs harbour a reductive domain covalently connected to a P450 domain and are known for their robust catalytic activity with great potential as biocatalysts. In an effort to expand genetic sources of self-sufficient CYPs, we devised a sequence-based screening system to identify them in a soil metagenome. Methods and Results: We constructed a soil metagenome library and performed sequence-based screening for self-sufficient CYP genes. A new CYP gene, syk181, was identified from the metagenome library. Phylogenetic analysis revealed that SYK181 formed a distinct phylogenic line with 46% amino-acid-sequence identity to CYP102A1 which has been extensively studied as a fatty acid hydroxylase. The heterologously expressed SYK181 showed significant hydroxylase activity towards naphthalene and phenanthrene as well as towards fatty acids. Conclusions: Sequence-based screening of metagenome libraries is expected to be a useful approach for searching self-sufficient CYP genes. The translated product of syk181 shows self-sufficient hydroxylase activity towards fatty acids and aromatic compounds. Significance and Impact of the Study: SYK181 is the first self-sufficient CYP obtained directly from a metagenome library. The genetic and biochemical information on SYK181 are expected to be helpful for engineering self-sufficient CYPs with broader catalytic activities towards various substrates, which would be useful for bioconversion of natural products and biodegradation of organic chemicals. © 2007 The Authors.
DOI
10.1111/j.1365-2672.2006.03169.x
Appears in Collections:
자연과학대학 > 화학·나노과학전공 > Journal papers
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