title: Streptomyces coelicolor A3(2) CYP102 protein, a novel fatty acid hydroxylase encoded as a heme domain without an N-terminal redox partner.
abstract: The gene from Streptomyces coelicolor A3(2) encoding CYP102B1, a recently discovered CYP102 subfamily which exists solely as a single P450 heme domain, has been cloned, expressed in Escherichia coli, purified, characterized, and compared to its fusion protein family members. Purified reconstitution metabolism experiments with spinach ferredoxin, ferredoxin reductase, and NADPH revealed differences in the regio- and stereoselective metabolism of arachidonic acid compared to that of CYP102A1, exclusively producing 11,12-epoxyeicosa-5,8,14-trienoic acid in addition to the shared metabolites 18-hydroxy arachidonic acid and 14,15-epoxyeicosa-5,8,11-trienoic acid. Consequently, in order to elucidate the physiological function of CYP102B1, transposon mutagenesis was used to generate an S. coelicolor A3(2) strain lacking CYP102B1 activity and the phenotype was assessed.
authors.affiliation: Institute of Life Science and Swansea Medical School, Grove Building, Swansea University, Swansea SA2 8PP, United Kingdom. d.c.lamb@swansea.ac.uk
authors.forename: David C
authors.initials: DC
authors.lastname: Lamb
authors.forename: Li
authors.initials: L
authors.lastname: Lei
authors.forename: Bin
authors.initials: B
authors.lastname: Zhao
authors.forename: Hang
authors.initials: H
authors.lastname: Yuan
authors.forename: Colin J
authors.initials: CJ
authors.lastname: Jackson
authors.forename: Andrew G S
authors.initials: AG
authors.lastname: Warrilow
authors.forename: Tove
authors.initials: T
authors.lastname: Skaug
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Dyson
authors.forename: Eric S
authors.initials: ES
authors.lastname: Dawson
authors.forename: Steven L
authors.initials: SL
authors.lastname: Kelly
authors.forename: David L
authors.initials: DL
authors.lastname: Hachey
authors.forename: Michael R
authors.initials: MR
authors.lastname: Waterman
mesh_terms: D015126:8,11,14-Eicosatrienoic Acid / Q000031:analogs & derivatives / Q000378:metabolism; D016718:Arachidonic Acid / Q000378:metabolism; D003001:Cloning, Molecular; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D004251:DNA Transposable Elements; D004926:Escherichia coli / Q000235:genetics; D005287:Ferredoxin-NADP Reductase / Q000378:metabolism; D005288:Ferredoxins / Q000378:metabolism; D015870:Gene Expression; D006899:Mixed Function Oxygenases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D016254:Mutagenesis, Insertional; D009249:NADP / Q000378:metabolism; D048372:Streptomyces coelicolor / Q000201:enzymology* / Q000235:genetics; D013379:Substrate Specificity
chemical_list: D004251:DNA Transposable Elements; D005288:Ferredoxins; D016718:Arachidonic Acid; D009249:NADP; C022948:10-hydroxy-11,12-epoxyeicosa-5,8,14-trienoic acid; C046782:14,15-epoxy-5,8,11-eicosatrienoic acid; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; D005287:Ferredoxin-NADP Reductase; D015126:8,11,14-Eicosatrienoic Acid