来自茄病镰刀菌菌株KUFI-6N的环己酮1,2-单加氧酶的纯化与特性分析

Purification and characterization of cyclohexanone 1,2-monooxygenase from Exophiala jeanselmei strain KUFI-6N.

作者信息

Hasegawa Y, Nakai Y, Tokuyama T, Iwaki H

机构信息

Department of Biotechnology, Faculty of Engineering and High Technology Research Center, Kansai University, Suita, Japan.

出版信息

Biosci Biotechnol Biochem. 2000 Dec;64(12):2696-8. doi: 10.1271/bbb.64.2696.

DOI:10.1271/bbb.64.2696

PMID:11210139

Abstract

Baeyer-Villiger cyclohexanone 1,2-monooxygenase (CHMO) was purified 17.1-fold from cell extracts of the fungus Exophiala jeanselmei grown on cyclohexanol to electrophoretically homogeneity by serial chromatographies. The molecular mass of the native enzyme was approximately 74 kDa by gel filtration and SDS-PAGE. Some enzymic characterizations were studied. The NH2-terminal amino acid residues were Ala-Lys-Ser-Leu-Asp-Val-Leu-Ile-Val-Gly-Ala-Gly-Phe-Gly-Gly-Ile-Tyr-Gln-Leu-, with similarity to the bacterial CHMOs of FAD-binding and NADPH-dependent type Baeyer-Villiger monooxygenases.

摘要

通过连续色谱法，从在环己醇上生长的真菌耶氏外瓶霉的细胞提取物中纯化出1,2 - 环己酮单加氧酶（CHMO），纯化倍数达17.1倍，达到电泳纯。通过凝胶过滤和SDS - PAGE测定，天然酶的分子量约为74 kDa。对该酶的一些特性进行了研究。其氨基末端氨基酸残基序列为Ala - Lys - Ser - Leu - Asp - Val - Leu - Ile - Val - Gly - Ala - Gly - Phe - Gly - Gly - Ile - Tyr - Gln - Leu -，与FAD结合型和NADPH依赖型拜耳 - 维利格单加氧酶的细菌CHMO具有相似性。