Recombinant destabilase from the medicinal leech: Preparation and properties
- Authors: Kurdyumov A.S.1, Manuvera V.A.1,2, Akhaev D.N.3, Baskova I.P.3, Lazarev V.N.1,2
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Affiliations:
- Federal Research and Clinical Centre of Physical-Chemical Medicine
- Moscow Institute of Physics and Technology (State University)
- Department of Biology
- Issue: Vol 42, No 1 (2016)
- Pages: 42-52
- Section: Article
- URL: https://journals.rcsi.science/1068-1620/article/view/227827
- DOI: https://doi.org/10.1134/S1068162016010088
- ID: 227827
Cite item
Abstract
The procedure for obtaining an active recombinant destabilase from the medicinal leech in Escherichia coli cells was developed. The plasmids encoding an analogue of native destabilase, as well as the protein forms carrying polyhistidine sequence at the Cand/or N-terminus of the polypeptide were obtained during the work. The producing strains of different forms of the protein were constructed, the cultivation process was optimized. The conditions of renaturation of destabilase recombinant forms by dialysis and using chromatographic absorbent were selected. The muramidase activity towards cell walls of Micrococcus lysodeikticus bacferia and lytic activity towards E. coli were investigated. The dependence of pH and ionic strength of the solution on the activities was determined. The total antibacterial activity of destabilase towards E. coli was shown.
Keywords
About the authors
A. S. Kurdyumov
Federal Research and Clinical Centre of Physical-Chemical Medicine
Author for correspondence.
Email: aleks-kuzmaland@bk.ru
Russian Federation, FMBA, ul. Malaya Pirogovskaya 1a, Moscow, 119435
V. A. Manuvera
Federal Research and Clinical Centre of Physical-Chemical Medicine; Moscow Institute of Physics and Technology (State University)
Email: aleks-kuzmaland@bk.ru
Russian Federation, FMBA, ul. Malaya Pirogovskaya 1a, Moscow, 119435; Institutskii per. 9, Dolgoprudny, Moscow region, 141700
D. N. Akhaev
Department of Biology
Email: aleks-kuzmaland@bk.ru
Russian Federation, Leninskie gory 1, Moscow, 119234
I. P. Baskova
Department of Biology
Email: aleks-kuzmaland@bk.ru
Russian Federation, Leninskie gory 1, Moscow, 119234
V. N. Lazarev
Federal Research and Clinical Centre of Physical-Chemical Medicine; Moscow Institute of Physics and Technology (State University)
Email: aleks-kuzmaland@bk.ru
Russian Federation, FMBA, ul. Malaya Pirogovskaya 1a, Moscow, 119435; Institutskii per. 9, Dolgoprudny, Moscow region, 141700