Purification and preliminary characterization of a cold-adapted extracellular proteinase from Trichoderma atroviride

L. Kredics, Kata Terecskei, Zsuzsanna Antal, A. Szekeres, L. Hatvani, L. Manczinger, Cs Vágvölgyi

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Eleven cold-tolerant Trichoderma isolates were screened for the production of proteolytic activities at 10°C. Based on the activity profiles determined with paranitroanilide substrates at 5 deg;C, strain T221 identified as Trichoderma atroviride was selected for further investigations. The culture broth of the strain grown at 10°C in casein-containing culture medium was concentrated by lyophilization and subjected to gel filtration, which was followed by chromatofocusing of the fraction showing the highest activity on N-benzoyl-Phe-Val-Arg-paranitroanilide. The purified enzyme had a molecular weight of 24 kDa, an isoelectric point of 7.3 and a pH optimum of 6.2. The temperature optimum of 25°C and the low thermal stability suggested that it is a true cold-adapted enzyme. Substrate specificity data indicate that the enzyme is a proteinase with a preference for Arg or Lys at the P1 position. The effect of proteinase inhibitors suggests that the enzyme has a binding pocket similar to the one present in trypsin.

Original languageEnglish
Pages (from-to)259-268
Number of pages10
JournalActa biologica Hungarica
Volume59
Issue number2
DOIs
Publication statusPublished - Jun 1 2008

Keywords

  • Cold tolerance
  • Psychrophilic enzymes
  • Trichoderma
  • Trypsin-like proteinase

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Environmental Science(all)
  • Neurology

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