RRC ID |
66227
|
著者 |
Yamamoto T, Kobayashi K, Hasegawa Y, Iwaki H.
|
タイトル |
Cloning, expression, and characterization of Baeyer-Villiger monooxygenases from eukaryotic Exophiala jeanselmei strain KUFI-6N.
|
ジャーナル |
Biosci Biotechnol Biochem
|
Abstract |
The fungus Exophiala jeanselmei strain 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 radicicola ATCC 11011, with 54.2% amino acid identity. ExCAMO was functionally expressed in E. coli and 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.
|
巻・号 |
85(7)
|
ページ |
1675-1685
|
公開日 |
2021-6-24
|
DOI |
10.1093/bbb/zbab079
|
PII |
6261182
|
PMID |
33930112
|
MeSH |
Cloning, Molecular
Enzyme Stability
Escherichia coli / genetics
Exophiala / enzymology*
Kinetics
Mixed Function Oxygenases / chemistry
Mixed Function Oxygenases / genetics*
Mixed Function Oxygenases / metabolism
Molecular Weight
Substrate Specificity
Temperature
|
IF |
1.516
|
リソース情報 |
原核生物(大腸菌) |
|