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

机译:水稻P450还原酶差异影响细菌表达系统中P450介导的代谢

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摘要

We describe cloning and characterization of three rice (Oryza sativa) NADPH-cytochrome P450 reductases (OsCPRs; E.C.I.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.
机译:我们描述了三种水稻(Oryza sativa)NADPH-细胞色素P450还原酶(OsCPRs; ECI6.2.4)的克隆和鉴定,它们是植物P450的潜在供体,包括血清素合成中的色胺5-羟化酶(T5H)和肉桂酸4-羟化酶( C4H)的合成。所有三个OsCPR转录本都在不同程度上受到压力的诱导。全长OsCPR1,OsCPR2和OsCPR3与T5H或C4H在大肠杆菌中的共表达表明OsCPR2 / T5H和OsCPR2 / C4H构建体显示出最高的T5H和C4H催化活性。即使具有短N末端缺失的缺失突变体能够还原细胞色素c,OsCPR2的N末端残基也是向P450传递电子的最大活性所必需的。

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