Cloning and expression of NDM-1 metallo-β-lactamase gene and study of the catalytic properties of the recombinant enzyme
- Авторлар: Grigorenko V.G.1, Rubtsova M.Y.1, Filatova E.V.1, Andreeva I.P.1, Mistryukova E.A.1, Egorov A.M.1
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Мекемелер:
- Department of Chemistry
- Шығарылым: Том 71, № 2 (2016)
- Беттер: 104-109
- Бөлім: Article
- URL: https://bakhtiniada.ru/0027-1314/article/view/163191
- DOI: https://doi.org/10.3103/S0027131416020048
- ID: 163191
Дәйексөз келтіру
Аннотация
A gene-expression system has been designed to express the NDM-1 metallo-β-lactamase gene in E. coli cells. This system enables the synthesis of the recombinant protein in a soluble and active form. A method for the isolation and purification of the recombinant enzyme has been developed. The yield of the homogeneous protein preparation was 10–15 mg per liter of E. coli culture medium. The catalytic parameters of the recombinant NDM-1 β-lactamase were measured for ampicillin (Km = 185 μM and kcat = 585 s–1) and meropenem (Km = 85 μM and kcat = 160 s–1). These values correlate well with the literature data. The catalytic parameters for the chromogenic CENTA substrate (Km = 14 μM and kcat = 290 s–1) were obtained for the first time.
Негізгі сөздер
Авторлар туралы
V. Grigorenko
Department of Chemistry
Хат алмасуға жауапты Автор.
Email: vitaly.grigorenko@gmail.com
Ресей, Moscow, 119991
M. Rubtsova
Department of Chemistry
Email: vitaly.grigorenko@gmail.com
Ресей, Moscow, 119991
E. Filatova
Department of Chemistry
Email: vitaly.grigorenko@gmail.com
Ресей, Moscow, 119991
I. Andreeva
Department of Chemistry
Email: vitaly.grigorenko@gmail.com
Ресей, Moscow, 119991
E. Mistryukova
Department of Chemistry
Email: vitaly.grigorenko@gmail.com
Ресей, Moscow, 119991
A. Egorov
Department of Chemistry
Email: vitaly.grigorenko@gmail.com
Ресей, Moscow, 119991
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