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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
Article

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 °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.

Keywords

Cold tolerance psychrophilic enzymes Trichoderma trypsin-like proteinase 

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Notes

Acknowledgements

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 (http://creativecommons.org/licenses/by/4.0/), 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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