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Recombinant Extracellular Cholesterol Oxidase from Nocardioides simplexстатья
- Авторы: Fokina Victoria V., Karpov Mikhail V., Kollerov Vyacheslav V., Bragin Eugeny Yu, Epiktetov Dmitry O., Sviridov Alexey V., Kazantsev Alexey V., Shutov Andrey A., Donova Marina V.
- Журнал: Biochemistry (Moscow)
- Том: 87
- Номер: 9
- Год издания: 2022
- Издательство: Pleiades Publishing, Ltd
- Местоположение издательства: Road Town, United Kingdom
- Первая страница: 903
- Последняя страница: 915
- DOI: 10.1134/s0006297922090048
- Аннотация: Cholesterol oxidase is a highly demanded enzyme used in medicine, pharmacy, agriculture, chemistry, and bio-technology. It catalyzes oxidation of 3β-hydroxy-5-ene- to 3-keto-4-ene- steroids with the formation of hydrogen perox-ide. Here, we expressed 6xHis-tagged mature form of the extracellular cholesterol oxidase (ChO) from the actinobacterium Nocardioides simplex VKM Ac-2033D (55.6 kDa) in Escherichia coli cells. The recombinant enzyme (ChO Ns) was purified using affinity chromatography. ChONs proved to be functional towards cholesterol, cholestanol, phytosterol, pregnenolone,and dehydroepiandrosterone. Its activity depended on the structure and length of the aliphatic side chain at C17 atom ofthe steroid nucleus and was lower with pregnenolone and dehydroepiandrosterone. The enzyme was active in a pH range of 5.25÷6.5 with the pH optimum at 6.0. Kinetic assays and storage stability tests demonstrated that the characteristics of ChONs were generally comparable with or superior to those of commercial ChO from Streptomyces hygroscopicus (ChOSh).The results contribute to the knowledge on microbial ChOs and evidence that ChO from N. simplex VKM Ac-2033D is a promising agent for further applications
- Добавил в систему: Казанцев Алексей Витальевич
Прикрепленные файлы
| № | Имя | Описание | Имя файла | Размер | Добавлен |
|---|---|---|---|---|---|
| 1. | Полный текст | BCM0903_Fokina.pdf | 2,2 МБ | 17 сентября 2022 [KazantsevAV] |