Email this article GENETIC DETERMINANTS OF SECRETED LYSOZYME INHIBITORS AND ENTEROBACTERIAANTI-LYSOZYME PHENOTYPE
- Authors: Perunova NB1, Andryushchenko SV1, Bukharin OV1, Perunova NB2, Andryuschenko SV2, Bukharin OV2
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Affiliations:
- Research Institute of Cellular and Intracellular Symbiosis, Orenburg, Russia
- Issue: Vol 89, No 6 (2012)
- Pages: 8-12
- Section: Articles
- URL: https://microbiol.crie.ru/jour/article/view/13691
- ID: 13691
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Abstract
anti-lysozyme phenotype. Materials and methods. 50 cultures of non-hemolytic Escherichia
coli, Klebsiella pneumoniae isolated from feces of patients with stage I - II intestine dysbiosis and Salmonella
enterica isolated from patients with acute intestinal infections (AII) and reconvalescent salmonella carriers
served as material for the study. Isolation of matrix DNA of the studied strains for PCR was carried out by
DNA-Express kit (Lytex Co., Russia). PCR screening of secreted lysozyme inhibitors genes ivyC and pliC
was performed based on Syntol Co. (Russia) primers and reagents. Amplicon detection was carried out by
agarose gel electrophoresis method. Detection of general anti-lysozyme activity (ALA) was performed by
plate method, secreted and sorption ALA components - photometrically by O.V.Bukharin et al. (1999) and
V.Yu.Sokolov and A.P.Luda (1987) methods, respectively. Results. Screening of chromosome localization
genes - ivyC in escherichiae and klebsiellae and pliC in salmonellae by PCR method showed their ubiquitous
spread among the studied cultures. The search for homologues of pliC gene that are characteristic for K.
pneumoniae virulence plasmid revealed positive result in 19±.5% of cases. The lowest level of ALA was
noted in carriers of only ivyC gene accounting for an average of 2.8±.6 g/ml.OD, while all enterobacteria
with pliC gene had average ALA value of 4±.3 g/ml.OD. Conclusion. Anti-lysozyme activity of enterobacteria
is part of the general mechanism of microorganism lysozyme resistance and is determined by the presence
of secreted lysozyme inhibitor genes. Application of molecular-genetic approach in the form of the
developed experimental test-system based on PCR method allowed to detect secreted lysozyme inhibitor
genes in screening mode. High level of secretory ALA is coupled with the presence of pliC gene, while in
ivyC gene carriers lower values of the attribute were detected.
About the authors
N B Perunova
S V Andryushchenko
O V Bukharin
N B Perunova
Research Institute of Cellular and Intracellular Symbiosis, Orenburg, RussiaResearch Institute of Cellular and Intracellular Symbiosis, Orenburg, Russia
S V Andryuschenko
Research Institute of Cellular and Intracellular Symbiosis, Orenburg, RussiaResearch Institute of Cellular and Intracellular Symbiosis, Orenburg, Russia
O V Bukharin
Research Institute of Cellular and Intracellular Symbiosis, Orenburg, RussiaResearch Institute of Cellular and Intracellular Symbiosis, Orenburg, Russia
References
- Андрющенко С.В. , Бухарин О.В., Перунова Н.Б., Иванова Е.В. Увеличение копийности плазмидной ДНК как возможный механизм усиления антилизоцимного признака бактерий под действием хлорамфеникола. Журн. микробиол. 2011, 6: 8-13.
- Бондаренко В.М., Петровская В.Г., Яблочков А.Л. Антилизоцимный фактор Klebsiella pneumoniae: природа, биологические функции и генетический контроль. Журн. микробиол. 1994, 1: 22-28.
- Бухарин О.В. Персистенция патогенных бактерий. М., Медицина, 1999.
- Бухарин О.В., Кириллов Д.А., Кириллов В.А. Скрининг плазмид у бактерий рода Bacillus, обладающих антилизоцимной активностью. Журн. микробиол. 2006, 1: 3-6.
- Маниатис Т., Фрич Э., Сэмбрук Дж. Методы генетической инженерии. Молекулярное клонирование. М., Мир, 1984.
- Abergel C., Monchois V., Byrne D. et al. Structure and evolution of the Ivy protein family, unexpected lysozyme inhibitors in gram-negative bacteria. Proc. Natl. Acad. Sci. USA. 2007, 104: 6394-6399.
- Callewaert L., Aertsen A., Deckers D. et al. A new family of lysozyme inhibitors contributing to lysozyme tolerance in gram-negative bacteria. PLoS Pathog. 2008, 4: e1000019.
- Chen Y.T., Chang H.Y., Lai Y.C. et al. Sequencing and analysis of the large virulence plasmid pLVPK of Klebsiella pneumoniae CG43. Gene. 2004, 337: 189-198.
- Deckers D., Masschalck B., Aertsen A. et al. Periplasmic lysozyme inhibitor contributes to lysozyme resistance in Escherichia coli. Cell. Mol. Life Sci. 2004, 61 (10): 1229-1237.
- Hiroshi N. Molecular basis of bacterial outer membrane permeability revisited. Microbiol. Mol. Biol. Rev. 2003,67 (4): 593-656.
- Monchois V., Abergel C., Sturgis J. et al. Escherichia coli ykfE ORFan gene encodes a potent inhibitor of C-type lysozyme. J. Biol. Chem. 2001, 276: 18437-18441.
- Wu K.M., Li L.H., Yan J.J. et al. Genome sequencing and comparative analysis of Klebsiella pneumoniae NTUH-K2044, a strain causing liver abscess and meningitis. J. Bacteriol. 2009, 191 (14): 4492-4501.
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