Antibiotic-dependent selection of E. coli clones with increased chaperone activity for highly efficient production of full-length soluble new delhi metallo-beta-lactamase

Autor: A V Kolesnikov, A V Kozyr, I G Shemiakin, O. N. Krasavtseva, A. E. Khlyntseva, N M Luneva
Rok vydání: 2013
Předmět:
Zdroj: Molecular Genetics, Microbiology and Virology. 28:147-155
ISSN: 1934-841X
0891-4168
DOI: 10.3103/s0891416813040046
Popis: The spread of the New Delhi metallo-beta-lactamase (NDM-1), a plasmid-borne enzyme conferring bacterial resistance to any known beta-lactam antibiotics, represents the global health threat. There is an urgent need to develop the efficient NDM-1 inhibitors of various mode of action thereby necessitating structural studies of the enzyme as well as analysis of the secretion pathway and localization of the protein. The recombinant full-length NDM-1 is produced in E. coli in the inactive form and is mostly accumulated in the inclusion bodies. The secreted recombinant NDM-1 forms are several N-terminally truncated species. The robust expression system capable of high-level production of the full-length NDM-1 and derivatives thereof is required to obtain NDM-1 in the quantities necessary for drug discovery, diagnostics, and research purposes. Therefore, we developed a new system that utilizes antibiotic pressure to select E. coli producing increased quantity of soluble NDM-1 and showed that an increase in the NDM-1 solubility occurs in the bacterial clones producing increased amounts in the chaperones.
Databáze: OpenAIRE
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