Rice P450 reductases differentially affect P450-mediated metabolism in bacterial expression systems.
We describe cloning and characterization of three rice (Oryza sativa) NADPH-cytochrome P450 reductases (OsCPRs; E.C.1.6.2.4) that are potential donors to plant P450s, including tryptamine 5-hydroxylase (T5H) in serotonin synthesis and cinnamate 4-hydroxylase (C4H) in phenylpropanoid synthesis. All three OsCPR transcripts are induced to varying degrees by stresses. Co-expression of full-length OsCPR1, OsCPR2 and OsCPR3 with either T5H or C4H in E. coli indicated that the OsCPR2/T5H and OsCPR2/C4H constructs displayed the highest T5H and C4H catalytic activities. The N-terminal residues of OsCPR2 were required for peak electron transfer activity to P450 even though deletion mutants with short N-terminal deletions were capable of reducing cytochrome c.
Department of Biotechnology, Interdisciplinary Program of Bioenergy and Biomaterials, Bioenergy Research Center, Chonnam National University, Gwangju, 500-757, Republic of Korea.
Sangkyu
S
Park
Young-Soon
YS
Kim
Sanjeewa G
SG
Rupasinghe
Mary A
MA
Schuler
Kyoungwhan
K
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D001419:Bacteria / Q000378:metabolism*; D001709:Biotechnology / Q000379:methods; D003001:Cloning, Molecular; D003373:Coumaric Acids / Q000737:chemistry; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism*; D045304:Cytochromes c / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D017353:Gene Deletion; D005786:Gene Expression Regulation; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D012275:Oryza / Q000201:enzymology* / Q000235:genetics; D011422:Propionates; D011994:Recombinant Proteins / Q000378:metabolism; D012701:Serotonin / Q000378:metabolism; D050564:Trans-Cinnamate 4-Monooxygenase / Q000378:metabolism
D003373:Coumaric Acids; D007527:Isoenzymes; D011422:Propionates; D011994:Recombinant Proteins; D012701:Serotonin; D045304:Cytochromes c; D003577:Cytochrome P-450 Enzyme System; D050564:Trans-Cinnamate 4-Monooxygenase; C407366:tryptamine 5-hydroxylase; C495469:p-coumaric acid