Abstract
Bacillus intermedius ribonuclease (binase), which is known to exert a growth-stimulating effect at low concentrations and a genotoxic effect at high concentrations, loses these abilities completely after exposure to 100°C for 10 min, but retains approximately 95% of its catalytic activity and structural integrity. Other types of modification, such as photoinactivation and site-specific mutagenesis, gave rise to enzyme forms with unaltered structure but reduced (sometimes to trace amounts) catalytic activity. Genotoxicity was always proportional to the catalytic activity of the native enzyme, while a notable growth-stimulating effect could be exerted by enzymes with low activity. The loss of biological activity of thermoinactivated binase was related to the increase in the number of negatively charged groups on the enzyme surface, which led to a substantial decline in the adhesive properties of the enzyme.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Kolpakov, A.I. and Kupriyanova, F.G., Effect of Exogenously Added RNases on the Reproduction of Candida tropicalis, Mikrobiologiya, 1992, vol. 61, no. 6, pp. 969–974.
Kipenskaya, L.V., Kupriyanova-Ashina, F.G., Il'inskaya, O.N., Kolpakov, A.I., and Leshchinskaya, I.B., Effect Of Bacillus intermedius RNase Preparations with Different Catalytic Activities on the Reproduction of Escherichia coli and Bacillus subtilis, Mikrobiologiya, 1998, vol. 67, no. 2, pp. 165–168.
Leshchinskaya, I.B., Kupriyanova, F.G., Yakovlev, G.I., Kipenskaya, L.V., and Ilinskaya, O.N., Molecular Mechanisms of Growth Stimulating Effects of Bacillus intermedius RNase, Karadeniz J. Med. Sci., 1995, vol. 8, no. 4, pp. 218–219.
Ilinskaya, O.N., Ivanchenko, O.N., Karamova, N.S., and Kipenskaya, L.V., SOS-inducing Ability of Native and Mutant Microbial Ribonucleases, Mutat. Res., 1996, vol. 354, pp. 203–209.
Kurinenko, B.M., Golubenko, I.A., Bulgakova, R.Sh., Davydov, R.E., Nekhoroshkova, Z.M., Valiullina, S.F., and Shlyapnikov, S.V., Photooxidation of Bacillus inter-medius RNase and Formation of Photoinactivated Product, Bioorg. Khimiya, 1986, vol. 12, no. 4, pp. 457–466.
Kupriyanova-Ashina, F.G., Dmitriev, V.V., Egorov, S.Yu., and Kolpakov, A.I., Penetration of Pancreatic DNase into Candida tropicalis Cells, Mikrobiologiya, 1992, vol. 61, no. 6, pp. 975–980.
Yakovlev, G.I., Moiseyev, G.P., Struminskaya, N.K., Borzykh, O.A., Kipenskaya, L.V., Znamenskaya, L.V., Leshchinskaya, I.B., Chernokalskaya, E.V., and Hartley, R.W., Mutation Analysis of Active Site of RNase Bacillus intermedius (Binase), FEBS Lett., 1995, vol. 354, pp. 305–306.
Randzhbar, B., Protasevich, I.I., Shul'ga, A.A., Kurbanov, F.T., Lobachev, V.M., Kirpichnikov, M.P., and Makarov, A.A., Barnase, Binase, and Their Hybrids: Differences in Conformation and Heat Denaturation Parameters, Mol. Biol. (Moscow), 1997, vol. 31, no. 3, pp. 492–499.
Balaban, N.P., Sharipova, F.R., Golubenko, I.A., et al., Physico chemical Characteristics of Ribonuclease Bacillus intermedius, Proc. 1st Int. Meeting “Structure and Chemistry of Ribonucleases,” Moscow, 1989, pp. 349–352.
Maron, M. and Ames, B.N., Revised Methods for the Salmonella Mutagenicity Test, Mutat. Res., 1983, vol. 113, pp. 173–215.
Qillardet, P. and Hofnung, M., The SOS Chromotest: A Review, Mutat. Res., 1993, vol. 297, pp. 235–279.
Kolpakov, A.I. and Il'inskaya, O.N., Optimized Assay of Ribonucleases with High-Polymeric RNA, Klin. Lab. Diagn., 1999, no. 5, pp. 14–16.
Protasevich, I.I., Platonov, A.L., Pavlovsky, A.G., and Esipova, N.G., Distribution of Charges in Bacillus intermedius 7P Ribonuclease Determines the Number of Cooperatively Melting Regions of the Globule, J. Biomol. Struct. Dyn., 1987, vol. 4, no. 5, pp. 885–893.
Faizullin, D.A. and Stupishina, E.A., Effect of Conformational Factors on the Reversibility of the Heat Denaturation of Binase, in Fermenty mikroorganizmov (Microbial Enzymes), Kazan, 1998, pp. 109–115.
Kupriyanova-Ashina, F.G., Lutskaya, A.Yu., Kolpakov, A.I., Kepecheva, R.M., and Leshchinskaya, I.B., Effect of Bacillus intermedius RNase on the Growth of Saccharomyces cerevisiae, Prikl. Biokhim. Mikrobiol., 1996, no. 2, pp. 254–259.
Joly, M., A Physico-Chemical Approach to the Denaturation of Proteins, London: Academic, 1965. Translated under the title Fizicheskaya khimiya denaturatsii belkov, Moscow: Mir, 1968.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kolpakov, A.I., Il'inskaya, O.N., Ivanchenko, O.B. et al. The Effect of Thermal Treatment on the Catalytic Activity and Biological Properties of Bacillus intermedius Ribonuclease. Microbiology 70, 18–21 (2001). https://doi.org/10.1023/A:1004828417874
Issue Date:
DOI: https://doi.org/10.1023/A:1004828417874