Usai Remigio, Kaluka Daniel, Mak Piotr J, Liu Yilin, Kincaid James R
Department of Chemistry, Marquette University, P.O. Box 1881, Milwaukee, WI 53201-1881, United States.
Department of Chemistry, Marquette University, P.O. Box 1881, Milwaukee, WI 53201-1881, United States.
J Inorg Biochem. 2020 Jul;208:111084. doi: 10.1016/j.jinorgbio.2020.111084. Epub 2020 May 5.
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.
细胞色素P450结合并裂解双氧以生成一种强效中间体化合物I，该化合物能够使惰性烃底物羟基化。细胞色素P450 119是一种细菌细胞色素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。尽管其天然底物和氧化还原伙伴仍然未知，但这种嗜热P450的一个潜在应用是将它们作为生物技术产业中的生物催化剂；例如，合成否则需要极端pH或温度等恶劣环境的有机化合物。在本工作中，通过冷冻辐解和共振拉曼光谱相结合的方法，研究了该酶与月桂酸（一种已知具有良好结合亲和力的替代底物）结合的氧化复合物，以捕获和表征关键瞬态酶中间体的活性位点结构，包括过氧和氢过氧物种。
