Streptomyces coelicolor A3(2) CYP102 protein, a novel fatty acid hydroxylase encoded as a heme domain without an N-terminal redox partner.
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.
Lamb
David C
DC
Institute of Life Science and Swansea Medical School, Grove Building, Swansea University, Swansea SA2 8PP, United Kingdom. d.c.lamb@swansea.ac.uk
Lei
Li
L
Zhao
Bin
B
Yuan
Hang
H
Jackson
Colin J
CJ
Warrilow
Andrew G S
AG
Skaug
Tove
T
Dyson
Paul J
PJ
Dawson
Eric S
ES
Kelly
Steven L
SL
Hachey
David L
DL
Waterman
Michael R
MR
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
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