ISSN Print 2500–1094
ISSN Online 2542–1204
BIOMEDICAL JOURNAL OF PIROGOV UNIVERSITY (MOSCOW, RUSSIA)
Copyright: © 2024 by the authors. Licensee: Pirogov University.
This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (CC BY).
This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (CC BY).
ORIGINAL RESEARCH
Complex antibacterial action of enzymes acting on Staphylococcus aureus biofilms
About authors
1 Sirius University of Science and Technology, Sirius, Sochi, Russia
2 Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, Pushchino, Russia
Correspondence should be addressed: Maxim O. Nagornykh
Prospekt Nauki, 5, Pushchino, 142290, Russia; moc.liamg@rennabred
About paper
Funding: the study was supported by the program of the Ministry of Science and Higher Education of the Russian Federation (agreement No. 075-10-2021-113, unique project ID: RF----193021X0001).
Author contribution: Zagoskin AA, Mirzoyan RA — creating genetic constructs, сhromatographic purification of recombinant proteins; Rezvykh LF — creating genetic constructs; Zakharova MV, Mubarakshina EK — selection of condition for recombinant protein production, experiments on production involving various E. coli strains; Nagornykh MO — study concept, genetic construct design, manuscript writing; Ivanov RA — general management.
Received: 2024-11-25
Accepted: 2024-12-15
Published online: 2024-12-23
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Fig. 1. Scheme of the experimental study involving production of enzybiotic drugs and testing their antibacterial activity
Fig. 2. Strategy of obtaining recombinant proteins prone to aggregation via production in the form of the chimeric molecule, in which there is an auxiliary polypeptide increasing the target protein solubility at the N-terminus. In particular, PAGE electropherogram demonstrates the examples of the TIG auxiliary polypeptide effects on the DNAse I solubility (А) and MBP auxiliary polypeptide effects on the endilysin PM9 solubility (B)
Fig. 3. Catalytic activity of two endolysins against the Staphylococcus aureus bacterial cell wall. Both recombinant enzymes possess antibacterial activity disrupting the cell wall. Endolysin LysK having a full-fledged catalytic domain possess significantly higher antibacterial activity against the gram-positive Staphylococcus aureus pathogen compared to endolysin PM9 having a truncated catalytic domain
Fig. 4. Destruction of the Staphylococcus aureus bacterial biofilms with the recombinant nuclease drugs and the combination drugs combining different catalytic activities (endolysin + nuclease). Nonspecific nucleases show higher activity in terms of bacterial biofilm disruption compared to DNAse I. Combining the enzymes showing different specificity in the same drug enhances antibacterial effects
Table 1. Estimation of the recombinant endolysin and nuclease production in E. coli strains
Note: S — soluble, I — insoluble, “+” — extent of soluble protein production.
Table 2. Estimation of the recombinant endolysin and nuclease production in E. coli strains
Note: S — soluble, I — insoluble, “+” — extent of soluble protein production.
