SSN 2410-7751 (Print)
ISSN 2410-776X (Online)
iotechnologia Acta Т. 15, No. 1, 2022
P. 72-80. Bibliography 22, Engl.
UDC: 579.6:604
https://doi.org/10.15407/biotech15.01.072
T. S. Todosiichuk 1, O. V. Pokas 2, S. О. Soloviov 3, T. S. Ryzhkova 1
1Igor Sikorsky Kyiv Polytechnic Institute, Kyiv
2SI “L.V. Gromashevsky Institute of Epidemiology and Infectious Diseases of NAMS of Ukraine”, Kyiv
3 Shupyk National Healthcare University of Ukraine, Kyiv
Aim. Evaluation of biotechnological aspects of the production of a liquid formulation of the multifunctional antiseptic preparation of microbial origin, which can be typified as an enzybiotic; characterization of the enzyme specificity of the studied formulation, stabilization methods, its ability to affect microbial biofilms.
Methods. Gel-filtration and electrophoresis were used to study the component composition and the specificity of the enzyme complex of the Streptomyces albus UN 44 producer strain. Proteolytic and staphylolytic activities of individual fractions were determined. The Pseudomonas aeruginosa biofilm and its formation under the effect of various drug formulations were quantitatively evaluated by spectrophotometry.
Results. The stability of the liquid formulation of the enzybiotic biosynthesized by S. albus UN 44 was demonstrated. Activity of the formulation could be prolonged and additionally stabilized by adding of 0.5% polyvinyl alcohol. Fractionation of the formulation enzyme complex using gel-filtration revealed the presence of at least three proteinases of different molecular weights (80-100, 24-35 and 20 kDa) and lysoenzymes (18-22 kDa). The effectiveness of the developed liquid antibiotic formulation for the destruction and inhibition (8-10 folds) of the biofilm formation by clinical strains of P. aeruginosa was shown.
Conclusions. The broad spectrum, multidirectional mechanisms of antimicrobial and regenerative action of enzybiotic drug, and the possibility of its production directly from the biotechnological process determine the prospects of its manufacturing and use as a multifunctional surface antiseptic.
Key words: antibiotic, antimicrobial action, Streptomyces albus, liquid formulation, biofilms, pathogens.
© Palladin Institute of Biochemistry of the National Academy of Sciences of Ukraine, 2022
References
1. P?rv?nescu H., B?l??oiu M., Ciurea M. E., B?l??oiu A. T., M?nescu R. Wound infections with multi-drug resistant bacteria. Chirurgia (Bucur). 2014, 109 (1), 73–79. https://www.revistachirurgia.ro/pdfs/2014-1-73.pdf
2. Valencia I. C., Kirsner R. S., Kerdel F. A. Microbiologic evaluation of skin wounds: alarmingtrendtowardantibioticresistanceinaninpatientdermatologyserviceduring a 10-year period. J. Am. Acad. Dermatol. 2004, 50 (6), 845–849. https://doi.org/10.1016/j.jaad.2003.11.064
3. Donlan R. M., Costerton J. W. Biofilms: survival mechanisms of clinically relevant microorganisms. Clin. Microbiol. Rev. 2002, 15 (2), 167–193. https://doi.org/10.1128/CMR.15.2.167-193.2002
4. Negut I., Grumezescu V., Grumezescu A. M. Treatment Strategies for Infected Wounds. Molecules. 2018, 23 (9), 2392. https://doi.org/10.3390/molecules23092392
5. Pilar R. F., Emaneini M., Beigverdi R., Banar M., van Leeuwen B. W., Jabalameli F. Combinatorial effects of antibiotics and enzymes against dual-species Staphylococcus aureus and Pseudomonasae ruginosa biofilms in the wound-like medium. PloS One. 2020, 15 (6). https://doi.org/10.1371/journal.pone.0235093
6. Nordmann P., Naas T., Fortineau N., Poirel L. Superbugs in the coming new decade; multidrug resistance and prospects for treatment of Staphylococcusaureus, Enterococcus spp. And Pseudomonasae ruginosa in 2010. Curr. Opin. Microbiol. 2007, 10 (5), 436–440. https://doi.org/10.1016/j.mib.2007.07.004
7. Xu G., Zhao Y., Du L. Hfq regulates antibacterial antibiotic biosynthesis and extra cellular lytic-enzyme production in Lysobacter enzymogenes OH11. Microbiol. Biotechnol. 2015, 8 (3), 499–509. https://doi.org/10.1111/1751-7915.12246
8. Genilloud O. Current approaches to exploit actinomycetes as a source of novel natural products. J. Ind. Microbiol. Biotechnol. 2011, 38 (3), 375–389. https://doi.org/10.1007/s10295-010-0882-7
9. Gurung N., Ray S., Bose S., Vivek R. A Broader View: Microbial Enzymes and Their Relevance in Industries, Medicine, and Beyond. Biomed. Res. Int. 2013, V. 2013, P. 1–18.https://doi.org/10.1155/2013/329121
10. Chakraborty A. K. Enzybiotics, A New Class of Enzyme Antimicrobials Targeted against Multidrug-Resistant Superbugs. Nov. Appro. Drug. Des. Dev. 2017, 2 (4), 1–4. https://doi.org/10.19080/NAPDD.2017.01.555576
11. Tiwari R., Dhama K., Chakraborty S., Kapoor S. Enzybiotics: New Weapon in the Army of Antimicrobials: A Review. Asian. J. Anim. Vet. Adv. 2014, 9 (3), 144–163. https://doi.org/10.3923/ajava.2014.144.163
12. S?o-Jos? C. Engineering of Phage-Derived Lytic Enzymes: Improving Their Potentialas Antimicrobials. Antibiotics. 2018, 7 (2), 29. https://doi.org/10.3390/antibiotics7020029
13. Sudhakar G. K., Kamath V., Pai A. Enzybiotics – A Review. Int. J. Pharmacol. Res. 2013, 3 (14), 69–71. https://www.researchgate.net/publication/307681509_Enzybiotics-_A_Review
14. Zhernosekova I. V., Sokolova I. E., Kilochek T. P. Characteristic of bacteriolytic enzyme complex of Streptomyces recifensisvar. lyticus 2Р-15. Bulletin of the Institute of Agricultural Microbiology. 2000, V. 7, P. 31–32. (In Russian).
15. Todosiichuk Т. S., Pokas О. V. Specificity analysis of finished forms of antimicrobial drugs from Streptomyce salbus. Eastern European Journal of Advanced Technology. 2015, 4/6 (76), 58–61. (In Ukrainian). https://doi.org/10.15587/1729-4061.2015.47730
16. Todosiichuk T. S., Klochko V. V., Savchuk Ya. I., Kobzysta O. P. New antibiotic substances of the Streptomyce salbus enzybiotic complex. Microbiol. J. 2019, 81 (5), 62–72.https://doi.org/10.15407/microbiolj81.05.062
17. Todosiichuk T. S. The development of technology of hydrolytic enzyme preparation Cytorecifen. Ph. D. dissertation. National technical university “KSh”, National University of food technologies. Kyiv, 2000.
18. Pavlova I. N., Zholner L. G., Zakharova I. Ya. Serine proteinase with lytic properties. Microbiology. 1988, 57 (3), 398 – 404.
19. Petrova I. S., Vintsyunaite M. N. Determination of the lytic and proteolytic activity of enzyme preparations of microbialorigin. Prikl. Biochem. Microbiol.1966, V. 2, P. 322 – 327.
20. Romanova Yu. M., Alekseeva N. V., Smirnova T. A. Ability to form biofilms in artificial systems in various strains of Salmonella typhymurium. Zhurn. Microbiol. 2006, V. 4, P. 38–42.
21. Rios Colombo N. S., Chalon M. C., Navarro S. A., Bellomio A. Pediocin-like bacteriocins: new perspectives on mechanism of action and immunity. Curr. Genet. 2017, 64 (2), 27–34. https://doi.org/10.1007/s00294-017-0757-9
22. Pokas O. V., Polishchuk O. I., Todosiichuk T. S. Influence of the enzyme preparation "Cytorecifen-M" for the ability of formation the biofilms by Pseudomonasae ruginosa strains. Preventive medicine. 2011, 2 (14), 81–85.