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

Kredics, L. and Terecskei, Kata and Antal, Zsuzsanna and Szekeres, A. and Hatvani, L. and Manczinger, L. and Vágvölgyi, Cs. (2008) Purification and preliminary characterization of a cold-adapted extracellular proteinase from Trichoderma atroviride. Acta Biologica Hungarica, 59 (2). pp. 259-268. ISSN 0236-5383

Text abiol.59.2008.2.11.pdf Restricted to Repository staff only until 30 June 2028. Download (513kB)

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 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.

Actions (login required)

Edit Item