Acta Biologica Hungarica

, Volume 59, Issue 2, pp 259–268 | Cite as

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. VágvölgyiEmail author


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 °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 JV-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.


Cold tolerance psychrophilic enzymes Trichoderma trypsin-like proteinase 


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We thank Miss Mária Lele for her technical help. L. Kredics is a grantee of the János Bolyai Research Scholarship.


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© Akadémiai Kiadó, Budapest 2008

This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (, which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

Authors and Affiliations

  • L. Kredics
    • 1
  • Kata Terecskei
    • 1
  • Zsuzsanna Antal
    • 1
  • A. Szekeres
    • 2
  • L. Hatvani
    • 1
  • L. Manczinger
    • 1
  • Cs. Vágvölgyi
    • 1
    Email author
  1. 1.Department of Microbiology, Faculty of Sciences and InformaticsUniversity of SzegedSzegedHungary
  2. 2.Cereal Research Non-profit CompanySzegedHungary

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