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

B. S. Kim, S. Y. Kim, J. Park, W. Park, K. Y. Hwang, Y. J. Yoon, W. K. Oh, B. Y. Kim, J. S. Ahn

Research output: Contribution to journalArticle

20 Citations (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.

Original languageEnglish
Pages (from-to)1392-1400
Number of pages9
JournalJournal of Applied Microbiology
Volume102
Issue number5
DOIs
Publication statusPublished - 2007 May

Keywords

  • Fatty acid hydroxylase
  • Metagenome
  • Self-sufficient cytochrome P450 monooxygenase
  • Sequence-based screening

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology

Fingerprint Dive into the research topics of 'Sequence-based screening for self-sufficient P450 monooxygenase from a metagenome library'. Together they form a unique fingerprint.

  • Cite this