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Resonance Raman spectroscopic studies of peroxo and hydroperoxo intermediates in lauric acid (LA)-bound cytochrome P450 119.
Journal of Inorganic Biochemistry ( IF 3.8 ) Pub Date : 2020-05-05 , DOI: 10.1016/j.jinorgbio.2020.111084
Remigio Usai 1 , Daniel Kaluka 1 , Piotr J Mak 1 , Yilin Liu 1 , James R Kincaid 1
Affiliation  

Cytochromes P450 bind and cleave dioxygen to generate a potent intermediate compound I, capable of hydroxylating inert hydrocarbon substrates. Cytochrome P450 119, a bacterial cytochrome P450 that serves as a good model system for the study of the intermediate states in the P450 catalytic cycle. CYP119 is found in high temperature and sulfur rich environments. Though the natural substrate and redox partner are still unknown, a potential application of such thermophilic P450s is utilizing them as biocatalysts in biotechnological industry; e.g., the synthesis of organic compounds otherwise requiring hostile environments like extremes of pH or temperature. In the present work the oxygenated complex of this enzyme bound to lauric acid, a surrogate substrate known to have a good binding affinity, was studied by a combination of cryoradiolysis and resonance Raman spectroscopy, to trap and characterize active site structures of the key fleeting enzymatic intermediates, including the peroxo and hydroperoxo species.



中文翻译:

月桂酸(LA)结合的细胞色素P450 119中过氧和氢过氧中间体的共振拉曼光谱研究。

细胞色素P450结合并裂解双氧以生成有效的中间体化合物I,该化合物能够羟基化惰性烃底物。细胞色素P450 119,一种细菌细胞色素P450,可作为研究P450催化循环中间状态的良好模型系统。CYP119存在于高温和富硫环境中。尽管仍不清楚天然底物和氧化还原伙伴,但这种嗜热P450的潜在应用是将其用作生物技术工业中的生物催化剂。例如,有机化合物的合成,否则需要恶劣的环境,例如pH或温度的极限。在目前的工作中,该酶的氧化复合物与月桂酸结合,月桂酸是一种已知具有良好结合亲和力的替代底物,

更新日期:2020-05-05
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