Proteolytic Activity of Fungus Pleurotus Ostreatus Mycelial Culture in Deep-Liquid Cultivation

Authors

  • А.Д. Кульгавеня (A.D. Kulgavenya), аспирант Polessky State University, Pinsk, Republic of Belarus
  • В.Н. Никандров (V.N. Nikandrov), д-р биол. наук, профессор Polessky State University, Pinsk, Republic of Belarus

Keywords:

proteases, mycelial culture, extracellular proteases, protein substrates, pH-dependence of proteolysis, class-specific protease inhibitors

Abstract

It was stated that proteinases of Pleurotus mycelium homogenate cleaved gelatin at pH 2.8–11.0 (proteolysis maximums at 5.0–8.5 and 10.6), casein at pH 2.8–9.0 (maximums at 2.8, 4.0–5.0, and 6.2–8.2), and hemoglobin at pH 2.8–11.0 (maximums at 4.6–5.6 and 7.0–8.2). At 10–3 M concentration of group-specific inhibitors, gelatinolytic activity was moderately (38–61%) suppressed by diisopropylfluorophosphate and metal-binding reagents. The activity of ‘alkaline’ gelatinases was completely suppressed by diisopropylfluorophosphate, p-chloromercuribenzoate, and o-phenanthroline. Caseinolytic activity at pH 2.8 and 5.2 was completely inhibited by pepstatin and at pH 5.2 – by EDTA. At pH 7.6 the caseinolytic activity was completely suppressed by diisopropylfluorophosphate and p-chloromercuribenzoate. Gelatinolytic activity of extracellular proteinases appeared at pH 3–11 (proteolysis maximum at 5.8–10.6), their caseinolytic activity – only at pH 7.6–8.2, and hemoglobinolytic activity – in the range of pH 7.6–10.0 (maximum at 8.6–9.4). However, judging by the protein concentration, the activity of extracellular proteinases exceeds that of intracellular enzymes in some cases by an order of magnitude. According to the results of the inhibitory analysis, gelatinolytic and caseinolytic activities of the culture fluid are provided mainly by serine proteinases. However, gelatinolytic activity of intracellular proteinases at pH 9.2 and extracellular proteinases at pH 7.6 was weakly sensitive to the class-specific inhibitors used in this study. This suggests the existence of some other type of proteinases.

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Published

2020-06-23

Issue

Section

Biological sciences