| Autor: |
Yamamoto, Taisei, Kobayashi, Kento, Hasegawa, Yoshie, Iwaki, Hiroaki |
| Zdroj: |
Bioscience, Biotechnology, and Biochemistry; July 2021, Vol. 85 Issue: 7 p1675-1685, 11p |
| Abstrakt: |
The fungus Exophiala jeanselmeistrain KUFI-6N produces a unique cycloalkanone monooxygenase (ExCAMO) that displays an uncommon substrate spectrum of Baeyer–Villiger oxidation of 4-10-membered ring ketones. In this study, we aimed to identify and sequence the gene encoding ExCAMO from KUFI-6N and overexpress the gene in Escherichia coli. We found that the primary structure of ExCAMO is most closely related to the cycloalkanone monooxygenase from Cylindrocarpon radicicolaATCC 11011, with 54.2% amino acid identity. ExCAMO was functionally expressed in E. coliand its substrate spectrum and kinetic parameters were investigated. Substrate profiling indicated that ExCAMO is unusual among known Baeyer–Villiger monooxygenases owing to its ability to accept a variety of substrates, including C4-C12 membered ring ketones. ExCAMO has high affinity and catalytic efficiency toward cycloalkanones, the highest being toward cyclohexanone. Five other genes encoding Baeyer–Villiger monooxygenases were also cloned and expressed in E. coli.Graphical AbstractThe kinetic parameters of ExCAMO toward C5-C9 membered ring cycloalkanones. The bars represent the Kmvalues. Solid line, kcat; dashed line, catalytic efficiency (kcat/Km). |
| Databáze: |
Supplemental Index |
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