Binding modes of CYP106A2 redox partners determine differences in progesterone hydroxylation product patterns.
Binding modes of CYP106A2 redox partners determine differences in progesterone hydroxylation product patterns.
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DOI：
10.1038/s42003-018-0104-9
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发表时间：
2018
影响因子：
5.9
通讯作者：
Bernhardt R
中科院分区：
文献类型：
--
作者：
Sagadin T;Riehm JL;Milhim M;Hutter MC;Bernhardt R
细菌细胞色素P450（P450）的天然氧化还原伙伴大多是未知的。因此，用异源氧化还原配偶体进行底物转化;在来自巨大芽孢杆菌ATCC 13368的CYP 106 A2的情况下，牛肾上腺素还蛋白（Adx）和肾上腺素还蛋白还原酶（AdR）。我们的目的是通过测试11种不同的氧化还原伴侣组合来优化CYP 106 A2的氧化还原系统，以改善产物形成。我们发现，电子转移蛋白1（516-618）显示出主产物15β-羟孕酮的最高产率，并且此外，产生减少量的不需要的多羟基化副产物。分子蛋白质-蛋白质对接表明，这是由微妙的结构变化导致的两种氧化还原酶的替代结合模式。停流测量分析了CYP 106 A2的降低，并显示表观速率常数存在显著差异，支持这一结论。这项研究提供了第一次，我们的知识合理的解释不同的产品模式的细胞色素P450所造成的氧化还原伙伴的结合模式的差异。Tanja Sagadin等人表明，可以使用不同的氧化还原系统来调节细胞色素P450 CYP 106 A2转化孕酮的速率选择性和产率。他们筛选了11种氧化还原伴侣组合，并确定了可用于改善单羟基和多羟基产品生物技术生产的特定组合。
Natural redox partners of bacterial cytochrome P450s (P450s) are mostly unknown. Therefore, substrate conversions are performed with heterologous redox partners; in the case of CYP106A2 from Bacillus megaterium ATCC 13368, bovine adrenodoxin (Adx) and adrenodoxin reductase (AdR). Our aim was to optimize the redox system for CYP106A2 for improved product formation by testing 11 different combinations of redox partners. We found that electron transfer protein 1(516–618) showed the highest yield of the main product, 15β-hydroxyprogesterone, and, furthermore, produced a reduced amount of unwanted polyhydroxylated side products. Molecular protein–protein docking indicated that this is caused by subtle structural changes leading to alternative binding modes of both redox enzymes. Stopped-flow measurements analyzing the CYP106A2 reduction and showing substantial differences in the apparent rate constants supported this conclusion. The study provides for the first time to our knowledge rational explanations for differences in product patterns of a cytochrome P450 caused by difference in the binding mode of the redox partners. Tanja Sagadin et al. show that different redox systems can be used to tune the rate selectivity and yield of progesterone conversion by the cytochrome P450 CYP106A2. They screen 11 redox partner combinations and identify specific combinations that may be used to improve biotechnological production of mono- and polyhydroxylated products.
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影响因子：
14.9
作者：
Comeau, SR;Gatchell, DW;Camacho, CJ
通讯作者：
Camacho, CJ
影响因子：
2.8
作者：
Brixius-Anderko, Simone;Hannemann, Frank;Bernhardt, Rita
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影响因子：
5.5
作者：
Hess, Berk
通讯作者：
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影响因子：
4.1
作者：
BERG, A;RAFTER, JJ
通讯作者：
RAFTER, JJ
影响因子：
4.1
作者：
Bernhardt, Rita
通讯作者：
Bernhardt, Rita