TY - JOUR
T1 - Rice P450 reductases differentially affect P450-mediated metabolism in bacterial expression systems
AU - Park, Sangkyu
AU - Kim, Young Soon
AU - Rupasinghe, Sanjeewa G.
AU - Schuler, Mary A.
AU - Back, Kyoungwhan
N1 - Funding Information:
Acknowledgments This research was supported by grants from the Basic Science Research Program (2009-0087207), the Priority Research Centers Program (2012-0005857) as well as the Chonnam National University (2010).
PY - 2013/3
Y1 - 2013/3
N2 - 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.
AB - 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.
KW - 4-Coumaric acid
KW - Cinnamate 4-hydroxylase
KW - E. coli expression
KW - NADPH-dependent cytochrome P450 reductases
KW - Oryza sativa; serotonin synthesis
KW - Tryptamine 5-hydroxylase
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U2 - 10.1007/s00449-012-0787-0
DO - 10.1007/s00449-012-0787-0
M3 - Article
C2 - 23053415
AN - SCOPUS:84880570691
SN - 1615-7591
VL - 36
SP - 325
EP - 331
JO - Bioprocess and Biosystems Engineering
JF - Bioprocess and Biosystems Engineering
IS - 3
ER -