Rice P450 reductases differentially affect P450-mediated metabolism in bacterial expression systems

Sangkyu Park, Young Soon Kim, Sanjeewa G. Rupasinghe, Mary A. Schuler, Kyoungwhan Back

Research output: Contribution to journalArticlepeer-review

Abstract

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.

Original languageEnglish (US)
Pages (from-to)325-331
Number of pages7
JournalBioprocess and Biosystems Engineering
Volume36
Issue number3
DOIs
StatePublished - Mar 2013

Keywords

  • 4-Coumaric acid
  • Cinnamate 4-hydroxylase
  • E. coli expression
  • NADPH-dependent cytochrome P450 reductases
  • Oryza sativa; serotonin synthesis
  • Tryptamine 5-hydroxylase

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering

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