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Estimation of protease activity in soils at low temperatures by casein amendment and with substitution of buffer by demineralized water

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The aim of this work was to modify the method of Ladd and Buttler (1972), by substituting Tris–HCl buffer (pH 8.52) with demineralized water (DEMI H2O), in order to assess its suitability for measurement of casein-protease activity at pH levels close to those of real soil in H2O. Measurements were undertaken over a range of incubation temperatures from 3 to 49 °C. Testing was performed on one organic soil and two different mineral soils. The substitution of Tris–HCl buffer by DEMI H2O at 49 °C decreased casein-protease activity to 67.25% in mineral soil and to 53.76% in organic soil. With decreasing temperature casein-protease activity decreased the most in organic soil, i.e., 0.07% of original its value at 3 °C. The incubation period was extended to maximally 336 h at 3 °C to totally obtain >10.0% of L-tyrosine equivalents released at optimum or close to optimum temperature and pH conditions. The Q10 values of casein-protease activity measured after substituting Tris–HCl buffer with DEMI H2O were unexpectedly high. Between the temperatures of 3 and 49 °C Q10 ranged from 3.46 to 4.25, whereas between 3 and 25 °C Q10 ranged from 6.78 to 11.08. Therefore, the modified method of Ladd and Buttler (1972) presented can be used for measurement of soil casein-protease activity under pH conditions close to that of real soil pH and at an averaged soil temperatures measured in the field. This modification makes possible an expression of soil casein-protease activity potential – when being combined with measurements of casein-protease activity under optimum or close to optimum temperature and pH conditions, if high concentration of casein is present.

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Authors and Affiliations

  1. Department of Geology and Soil Science, Mendel University of Agriculture and Forestry, Brno, Czech Republic

    K. Rejsek & P. Formanek

  2. Laboratory of Ecological Physiology of Forest Trees, Institute of Systems Biology and Ecology, Brno, Czech Republic

    M. Pavelka

Authors
  1. K. Rejsek
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  2. P. Formanek
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  3. M. Pavelka
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Authors’ address: Klement Rejsek, Department of Geology and Soil Science, Mendel University of Agriculture and Forestry, Zemedelska 3, 613 00 Brno, Czech Republic

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Rejsek, K., Formanek, P. & Pavelka, M. Estimation of protease activity in soils at low temperatures by casein amendment and with substitution of buffer by demineralized water. Amino Acids 35, 411–417 (2008). https://doi.org/10.1007/s00726-007-0601-5

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  • DOI: https://doi.org/10.1007/s00726-007-0601-5

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