title: Unusually broad substrate profile of self-sufficient cytochrome P450 monooxygenase CYP116B4 from Labrenzia aggregata.
abstract: A new member of the CYP116B subfamily-P450LaMO-was discovered in Labrenzia aggregata by genomic data mining. It was successfully overexpressed in Escherichia coli, purified, and subsequently characterized spectroscopically, and its catalytic properties were assessed. Substrate profiling of the P450LaMO revealed that it was a versatile catalyst, exhibiting hydroxylation and epoxidation activities as well as O-dealkylation and asymmetric sulfoxidation activities. Diverse compounds, including alkylbenzenes, aromatic bicyclic molecules, and terpenoids, were shown to be hydroxylated by P450LaMO. Such diverse catalytic activities are uncommon for the bacterial P450s, and the P450LaMO-mediated stereoselective hydroxylation of inactivated C-H bonds-ubiquitous and relatively unreactive in organic molecules-is particularly unusual. The self-sufficient nature of P450LaMO, coupled with its broad substrate range, highlights it as an ideal template for directed evolution towards various applications.
authors.affiliation: Laboratory of Biocatalysis and Synthetic Biotechnology, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai 200237 (China).
authors.forename: Yue-Cai
authors.initials: YC
authors.lastname: Yin
authors.forename: Hui-Lei
authors.initials: HL
authors.lastname: Yu
authors.forename: Zheng-Jiao
authors.initials: ZJ
authors.lastname: Luan
authors.forename: Ren-Jie
authors.initials: RJ
authors.lastname: Li
authors.forename: Peng-Fei
authors.initials: PF
authors.lastname: Ouyang
authors.forename: Jing
authors.initials: J
authors.lastname: Liu
authors.forename: Jian-He
authors.initials: JH
authors.lastname: Xu
mesh_terms: D000595:Amino Acid Sequence; D055162:Biocatalysis; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D006900:Hydroxylation; D008969:Molecular Sequence Data; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D041902:Rhodobacteraceae / Q000201:enzymology*; D016415:Sequence Alignment; D013379:Substrate Specificity
chemical_list: D011994:Recombinant Proteins; D003577:Cytochrome P-450 Enzyme System
keywords: cytochromes; data mining; hydroxylation; oxidoreductases; protein engineering; substrate promiscuity