ANALYSIS OF OSMOTIC STABILITY OF BACTERIAL PROTOPLASTS IN FREE AND IMMOBILIZED STATES

UDC 612.51:631.461:576.8

  • Ignatenko Arkadiy Vasil’yevich – PhD (Biology), Associate Professor, Assistant Professor, the Department of Biotechnology. Belarusian State Technological University (13a, Sverdlova str., 220006, Minsk, Republic of Belarus). E-mail: ignatenko_av @tut.by

DOI: https://doi.org/10.52065/2520-2669-2022-253-1-58-65

Key words: bacteria, protoplasts, immobilization, agar, osmotic resistance, light scattering, biocalorimetry, bioluminescence, biosensor analysis.

For citation: Ignatenko A. V. Analysis of osmotic stability of bacterial protoplasts in free and immobilized states. Proceedings of BSTU, issue 2, Chemical Engineering, Biotechnologies, Geoecology, 2022, no. 1 (253), pp. 58–65 (In Russian). DOI: https://doi.org/10.52065/2520-2669-2022-253-1-58-65.

Abstract

The paper considers the resistance of protoplasts to osmolysis in a free and immobilized agar state by light scattering, biocalorimetry and bioluminescence methods in order to determine the possibility of using protoplasts for biosensory analysis of aqueous media. The study of changes in the light scattering of protoplasts of В. subtilis bacteria in physiological medium and from the sucrose concentration in the range of 0–30% showed that it is associated with both an increase in the size of protoplasts and a decrease in their concentration. Three stages of light scattering changes during protoplasts formation of В. subtilis bacteria were noted and discussed. The analysis of the osmotic stability of protoplasts in the immobilized state, carried out by the method of biocalorimetry, showed a 2-stage character of the change in their heat release when placed in physiological medium. This is explained from the standpoint of Selye’s stress theory as the stress and distress states of protoplasts. The analysis of osmolysis of protoplasts by biocalorimetry and bioluminescence showed similar results. Immobilization of protoplasts in agar showed that their osmotic stability increases, however, 0.7% agar did not provide sufficient osmotic stability and only 2% agar allows replacing the hypertonic medium with a gel environment, which allows the use of protoplasts as part of biosensor devices.

Download

References

  1. Yurkova I. L. Bioanalitika [Bioanalytics]. Minsk, BGU Publ., 2017. 359 p. (In Russian).
  2. Terner A., Karube I., Uilson Dzh. Biosensory: osnovy i prilozheniya [Biosensors: Fundamentals and Applications]. Moscow, Mir Publ., 1992. 614 p. (In Russian).
  3. Biologicheskiy kontrol’ okruzhayushchey sredy: bioindikatsiya i biotestirovaniye [Biological control of the environment: bioindication and biotesting]. Edit. by O. P. Melekhov, E. I. Sarapul’cev. Moscow, Akademiya Publ., 2010. 288 p. (In Russian).
  4. Kitova A. E., Alferov V. A., Ponamoreva O. N., Kuzmichev A. V., Ezhkov A. A., Arsen’ev D. V., Reshetilov A. N. Enzyme biosensors for express analysis of glucose, ethanol and starch content in fermentation media. Mikrobnyye biokatalizatory i perspektivy razvitiya fermentnykh tekhnologiy v pererabatyvayushchikh otraslyakh APK [Microbial biocatalysts and prospects for the development of enzyme technologies in the processing industries of agriculture]. Moscow, Pishchepromizdat Publ., 2004, pp. 255–262 (In Russian).
  5. Farmatsevticheskaya biotekhnologiya [Pharmaceutical biotechnology]. Edit. by D. V. Moiseev. Vitebsk, VGMU Publ., 2019. 293 p. (In Russian).
  6. Fedorova G. I. Properties of bacterial protoplasts and spheroplasts. Bull. Exp. Biol. Med., 1969, no. 68. pp. 66–69.
  7. Eliseeva I. V., Babich E. M., Volyanskij Yu. L., Sklyar N. I., Belozerskij V. I. A role of latent, difficultly cultivated or non-cultivated persistent bacteria in human pathology. Annals of Mechnikov’s Institute, 2006, no. 1, pp. 12–46.
  8. Yakovenko K. N., Troickiy N. A. Protoplasty mikroorganizmov [Protoplasts of microorganisms]. Minsk, Nauka i tekhnika Publ., 1985. 160 p. (In Russian).
  9. Ignatenko A. V. Bio-calorimetric analysis of the safety of aquatic environments using bacterial protolayers. Biotekhnologiya: vzglyad v budushchee: materialy III Mezhdunarodnoy nauchno-prakticheskoy konferentsii [Biotechnology: a look into the future: materials of the III International scientific and practical conference]. Stavropol’, 2017, pp. 267–271 (In Russian).
  10. Immobilizovannyye kletki i fermenty. Metody [Immobilized cells and enzymes. Methods]. Edit. by J. Woodward. Moscow, Mir Publ., 1988. 215 p. (In Russian).
  11. Ignatenko A. V. Mikrobiologicheskiye metody kontrolya kachestva pishchevykh produktov [Microbiological methods of food quality control], Minsk, BSTU Publ., 2015. Part 2. 202 p. (In Russian).
  12. Frolov Yu. G. Kurs kolloidnoy khimii [Colloidal chemistry]. Мoscow, Аl’yans Publ., 2004. 424 p. (In Russian).
  13. Antonov V. F., Chernysh A. M., Pasechnik V. I., Voznesenskij S. A., Kozlova E. K. Biofizika [Biophysics]. Moscow, Vlados Publ., 1999. 288 p. (In Russian).
24.11.2021