Cytochrome P450 CYP199A4 from Rhodopseudomonas palustris Catalyzes Heteroatom Dealkylations, Sulfoxidation, and Amide and Cyclic Hemiacetal Formation 机翻标题: 暂无翻译,请尝试点击翻译按钮。

ACS catalysis
2018 / 8 / 7
Univ Adelaide, Dept Chem, Adelaide, SA 5005, Australia;Univ Adelaide, Sch Biol Sci, Adelaide, SA 5005, Australia;Univ Queensland, Sch Chem & Mol Biosci, Brisbane, Qld 4072, Australia;Univ Adelaide, Dept Chem, Adelaide, SA 5005, Australia;Univ Adelaide, Dept Chem, Adelaide, SA 5005, Australia;Univ Queensland, Sch Chem & Mol Biosci, Brisbane, Qld 4072, Australia;
Podgorski, Matthew N.;Bruning, John B.;De Voss, James J.;Bell, Stephen G.;Coleman, Tom;Wong, Siew Hoon;
The cytochrome P450 enzymes execute a range of selective oxidative biotransformations across many biological systems. The bacterial enzyme CYP199A4 catalyzes the oxidative demethylation of 4-methoxybenzoic acid. The benzoic acid moiety of the molecule binds in the active site of the enzyme such that the functional group at the para-position is held close to the heme iron. Therefore, CYP199A4 has the potential to catalyze alternative monooxygenase reactions with different para-substituted benzoic acid substrates such as thioethers and alkylamines. The oxidation of 4-methyl- and 4-ethyl-thiobenzoic acids by CYP199A4 resulted in sulfur oxidation. 4-Ethylthiobenzoic acid sulfoxidation and 4-ethylbenzoic acid hydroxylation by CYP199A4 occurred with high enantioselectivity (>74% enantiomeric excess). By way of contrast, CYP199A4 catalyzed exclusive oxidative N-demethylation over N-oxide formation with 4-methyl- and 4-dimethylaminobenzoic acids. Unexpectedly acetamide formation by CYP199A4 competes with dealkylation in the turnover of 4-ethyl- and diethyl-aminobenzoic acids. No oxidative dealkylation was observed with 3,4-ethylenedioxybenzoic with only hydroxylation to form a cyclic hemiacetal being detected. The X-ray crystal structures of four substrate-bound forms of the enzyme were solved and revealed subtle changes in the location of the para substituent which, when combined with the reactivity of the substituents, provided a basis for understanding the changes in selectivity. Furthermore, in the 4-ethylthiobenzoic acid-bound structure, the active site residue Phe298 moves to accommodate the substituent which points away from the heme iron. As such, the CYP199A4 enzyme provides ready access to a combination of structural, binding, and activity data with which to study a variety of reactions which are catalyzed by the P450 superfamily of enzymes.
biocatalysis;cytochrome P450 enzymes;dealkylation;heteroatom oxidation;crystal structures;C-H bond oxidation;enzyme mechanism;