Cloning and expression of NDM-1 metallo-β-lactamase gene and study of the catalytic properties of the recombinant enzyme


Дәйексөз келтіру

Толық мәтін

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Рұқсат жабық Тек жазылушылар үшін

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.

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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