Investigating the roles of T224 and T232 in the oxidation of cinnamaldehyde catalyzed by myxobacterial CYP260B1
Investigating the roles of T224 and T232 in the oxidation of cinnamaldehyde catalyzed by myxobacterial CYP260B1
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DOI：
10.1002/1873-3468.12519
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发表时间：
2017-01-01
期刊：
影响因子：
3.5
通讯作者：
Khatri, Yogan
中科院分区：
文献类型：
--
作者：
Litzenburger, Martin;Lo Izzo, Roberta;Khatri, Yogan
虽然醛氧化成羧酸在自然界中主要由醛脱氢酶催化，但细胞色素P450也能够进行这样的反应。在这项研究中，我们证明了氧化肉桂醛肉桂酸的粘细菌CYP260 B1。在我们对醛的对接研究之后，我们通过定点诱变分别产生了CYP 260 B1的T224A和T234A突变体，以破坏底物定位和质子递送。此外，我们使用的动力学溶剂同位素效应的底物的稳态营业额调查的活性中间体能够进行催化。我们的研究结果表明，醛氧化发生通过亲核攻击的铁过氧阴离子。
Although the oxidation of aldehydes to carboxylic acids is mainly catalyzed by aldehyde dehydrogenases in nature, cytochromes P450 are also able to perform such reactions. In this study, we demonstrate the oxidation of cinnamaldehyde to cinnamic acid by the myxobacterial CYP260B1. Following our docking studies of the aldehyde, we generated T224A and T234A mutants of CYP260B1 by site-directed mutagenesis to disrupt the substrate positioning and proton delivery, respectively. Furthermore, we used the kinetic solvent isotope effect on the steady-state turnover of the substrate to investigate the reactive intermediate capable of performing the catalysis. Our results suggest that the aldehyde oxidation occurs via a nucleophilic attack of the ferric peroxoanion.