期刊
BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY
卷 69, 期 4, 页码 1741-1756出版社
WILEY
DOI: 10.1002/bab.2243
关键词
biocatalysis; cytochrome P450; enzyme; monooxygenases; peroxidation; protein engineering
资金
- Scientific and Technological Research Council of Turkey [116Z380]
By studying the variant DM CYP119 of acidothermophilic P450, it was found to have higher stability towards organic peroxides and higher peroxidation activity for non-natural substrates compared to wild-type CYP119. It also showed higher affinity for progesterone and other bioactive potential substrates, indicating its wide range of potential applications in the pharmaceutical and chemical industry.
Biocatalysts are increasingly applied in the pharmaceutical and chemical industry. Cytochrome P450 enzymes (P450s) are valuable biocatalysts due to their ability to hydroxylate unactivated carbon atoms using molecular oxygen. P450s catalyze reactions using nicotinamide adenine dinucleotide phosphate (NAD(P)H) cofactor and electron transfer proteins. Alternatively, P450s can utilize hydrogen peroxide (H2O2) as an oxidant, but this pathway is inefficient. P450s that show higher efficiency with peroxides are sought after in industrial applications. P450s from thermophilic organisms have more potential applications as they are stable toward high temperature, high and low pH, and organic solvents. CYP119 is an acidothermophilic P450 from Sulfolobus acidocaldarius. In our previous study, a novel T213R/T214I (double mutant [DM]) variant of CYP119 was obtained by screening a mutant library for higher peroxidation activity utilizing H2O2. Here, we characterized the substrate scope; stability toward peroxides; and temperature and organic solvent tolerance of DM CYP119 to identify its potential as an industrial biocatalyst. DM CYP119 displayed higher stability than wild-type (WT) CYP119 toward organic peroxides. It shows higher peroxidation activity for non-natural substrates and higher affinity for progesterone and other bioactive potential substrates compared to WT CYP119. DM CYP119 emerges as a new biocatalyst with a wide range of potential applications in the pharmaceutical and chemical industry.
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