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

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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; Bo Yeon KimJong Seog Ahn
Bibliographic Citation
Journal of Applied Microbiology, vol. 102, no. 5, pp. 1392-1400
Publication Year
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.
fatty acid hydroxylasemetagenomeself-sufficient cytochrome P450 monooxygenasesequence-based screening
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Ochang Branch Institute > Chemical Biology Research Center > 1. Journal Articles
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