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Purification and characterization of a subtilisin-like proteinases secreted in the stationary growth phase of Bacillus amyloliquefaciens H2

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Abstract

Proteinases secreted during the early and late stationary phases have been isolated from the culture liquid of Bacillus amyloliquefaciens H2 using CM-cellulose ion-exchange chromatography with subsequent FPLC on a Mono S column. Considering the character of hydrolysis of specific chromogenic substrates and the type of inhibition, these enzymes were identified as subtilisin-like proteinases. The molecular weight of both proteinases is 29 kD. The proteolytic activity of the proteinases secreted during the early and late stationary phases towards the synthetic substrate Z-Ala-Ala-Leu-pNA was maximal at pH 8.5 and 9.0, respectively. The maximal activity of both proteinases was observed at 37°C, and the proteins were stable within the pH range of 7.2–9.5. The subtilisin-like proteinases from B. amyloliquefaciens were shown to catalyze synthesis of peptide bonds.

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Authors and Affiliations

  1. Biological and Soil Science Faculty, Kazan State University, ul. Kremlevskaya 18, 420008, Kazan, Russia

    N. P. Balaban, L. A. Malikova, A. M. Mardanova & M. R. Sharipova

  2. Chemical Faculty, Lomonosov Moscow State University, 119899, Moscow, Russia

    G. N. Rudenskaya

Authors
  1. N. P. Balaban
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  2. L. A. Malikova
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  3. A. M. Mardanova
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  4. G. N. Rudenskaya
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  5. M. R. Sharipova
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Corresponding author

Correspondence to A. M. Mardanova.

Additional information

Original Russian Text © N. P. Balaban, L. A. Malikova, A. M. Mardanova, G. N. Rudenskaya, M. R. Sharipova, 2007, published in Biokhimiya, 2007, Vol. 72, No. 4, pp. 568–575.

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Balaban, N.P., Malikova, L.A., Mardanova, A.M. et al. Purification and characterization of a subtilisin-like proteinases secreted in the stationary growth phase of Bacillus amyloliquefaciens H2. Biochemistry Moscow 72, 459–465 (2007). https://doi.org/10.1134/S0006297907040141

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  • DOI: https://doi.org/10.1134/S0006297907040141

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