Bacteriophage enzymes for the prevention and treatment of bacterial infections: Stability and stabilization of the enzyme lysing Streptococcus pyogenes cells |
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Authors: | N. L. Klyachko N. F. Dmitrieva A. S. Eshchina O. V. Ignatenko L. Yu. Filatova E. I. Rainina A. K. Kazarov A. V. Levashov |
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Affiliation: | (1) Faculty of Chemistry, Moscow State University, Vorobyevy gory, Moscow, 119992, Russia;(2) Faculty of Preventive Medicine, Sechenov Moscow Medical Academy, Trubetskaya ul. 8-2, Moscow, 119992, Russia;(3) Pacific Northwest National Laboratory, P.O. Box 999, Richland, WA 99352, USA |
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Abstract: | The effect of various compounds on the activity and stability of a phage-associated enzyme lysing cells of streptococci of groups A and C (PlyC) was investigated. Substantial inhibition of the enzyme activity was revealed at an increased ionic strength (in the presence of NaCl) and upon the addition of carbohydrates (mono-, di-, and polysaccharides), i.e., agents stabilizing many enzymes. It was established that the enzyme activity was substantially reduced in the presence of positively charged polyelectrolytes and surfactants, whereas incubation with micelle-forming substances and negatively charged polyelectrolytes led to PlyC activation and stabilization. It was shown that, in the micellar polyelectrolyte composition M16, the enzyme retained its activity for 2 months; while in a buffer solution under the same conditions (pH 6.3, room temperature), ture), it practically completely lost its activity in 2 days. Characteristics of the enzyme thermal inactivation were found, in particular, its half-inactivation time at various temperatures; these allowed us to estimate its behavior at any temperature and to recommend conditions for its storage and use. |
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Keywords: | a phage-associated enzyme lysing streptococcal cells bacterial infections phage enzyme stability and stabilization phage enzymes streptococcal infections |
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