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  • 1 University of Szeged Department of Microbiology, Faculty of Sciences and Informatics Szeged Hungary
  • | 2 Cereal Research Non-profit Company Szeged Hungary
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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.

  • Antal, Z., Kredics, L., Manczinger, L., Ferenczy, L. (2001) Extracellular enzyme profiles of mycoparasitic Trichoderma strains. IOBC/WPRS Bull. 24 , 337–340.

    Ferenczy L. , 'Extracellular enzyme profiles of mycoparasitic Trichoderma strains ' (2001 ) 24 IOBC/WPRS Bull. : 337 -340.

    • Search Google Scholar
  • Antal, Z., Manczinger, L., Szakács, G., Tengerdy, R. P., Ferenczy, L. (2000) Colony growth, in vitro antagonism and secretion of extracellular enzymes in cold-tolerant strains of Trichoderma species. Mycol. Res. 104 , 545–549.

    Ferenczy L. , 'Colony growth, in vitro antagonism and secretion of extracellular enzymes in cold-tolerant strains of Trichoderma species ' (2000 ) 104 Mycol. Res. : 545 -549.

    • Search Google Scholar
  • Benítez, T., Rincón, A. M., Limón, M. C., Codón A. C. (2004) Biocontrol mechanisms of Trichoderma strains. Int. Microbiol. 7 , 249–260.

    Codón A. C. , 'Biocontrol mechanisms of Trichoderma strains ' (2004 ) 7 Int. Microbiol. : 249 -260.

    • Search Google Scholar
  • Clarkson, J. P., Mead, A., Payne, T., Whipps, J. M. (2004) Effect of environmental factors and Sclerotium cepivorum isolate on sclerotial degradation and biological control of white rot by Trichoderma. Plant Pathol. 53 , 353–362.

    Whipps J. M. , 'Effect of environmental factors and Sclerotium cepivorum isolate on sclerotial degradation and biological control of white rot by Trichoderma ' (2004 ) 53 Plant Pathol. : 353 -362.

    • Search Google Scholar
  • Druzhinina, I., Kopchinskiy, A. G., Komon, M., Bissett, J., Szakacs, G., Kubicek, C. P. (2005) An oligonucleotide barcode for species identification in Trichoderma and Hypocrea . Fungal Genet. Biol. 42 , 813–828.

    Kubicek C. P. , 'An oligonucleotide barcode for species identification in Trichoderma and Hypocrea ' (2005 ) 42 Fungal Genet. Biol. : 813 -828.

    • Search Google Scholar
  • Druzhinina, I., Kubicek, C. P. (2005) Species concepts and biodiversity in Trichoderma and Hypocrea : from aggregate species to species clusters? J. Zhejiang Univ. Sci. 6B , 100–112.

    Kubicek C. P. , 'Species concepts and biodiversity in Trichoderma and Hypocrea: from aggregate species to species clusters? ' (2005 ) 6B J. Zhejiang Univ. Sci. : 100 -112.

    • Search Google Scholar
  • Feller, G. (2003) Molecular adaptations to cold in psychrophilic enzymes. Cell. Mol. Life Sci. 60 , 648–662.

    Feller G. , 'Molecular adaptations to cold in psychrophilic enzymes ' (2003 ) 60 Cell. Mol. Life Sci. : 648 -662.

    • Search Google Scholar
  • Howell, C. R. (2003) Mechanisms employed by Trichoderma species in the biological control of plant diseases: the history and evolution of current concepts. Plant Dis. 87 , 4–10.

    Howell C. R. , 'Mechanisms employed by Trichoderma species in the biological control of plant diseases: the history and evolution of current concepts ' (2003 ) 87 Plant Dis. : 4 -10.

    • Search Google Scholar
  • Kredics, L., Antal, Z., Szekeres, A., Hatvani, L., Manczinger, L., Vágvölgyi, C., Nagy, E. (2005) Extracellular proteases of Trichoderma species — a review. Acta Microbiol. Immunol. Hung. 52 , 169–184.

    Nagy E. , 'Extracellular proteases of Trichoderma species — a review ' (2005 ) 52 Acta Microbiol. Immunol. Hung. : 169 -184.

    • Search Google Scholar
  • Kubicek, C. P., Mach, R. L., Peterbauer, C. K., Lorito, M. (2001) Trichoderma: from genes to biocontrol. J. Plant Pathol. 83(S2) , 11–23.

    Lorito M. , 'Trichoderma: from genes to biocontrol ' (2001 ) 83 J. Plant Pathol. : 11 -23.

  • Lieckfeldt, E., Kullnig, C. M., Kubicek, C. P., Samuels, G. J., Börner, T. (2001) Trichoderma aureoviride : phylogenetic position and characterization. Mycol. Res. 105 , 313–322.

    Börner T. , 'Trichoderma aureoviride: phylogenetic position and characterization ' (2001 ) 105 Mycol. Res. : 313 -322.

    • Search Google Scholar
  • Loveland, J., Gutshall, K., Kasmir, J., Prema, P., Brenchley, J. E. (1994) Characterisation of psychrotropic microorganisms producing β-galactosidase activities. Appl. Environ. Microbiol. 60 , 12–28.

    Brenchley J. E. , 'Characterisation of psychrotropic microorganisms producing β-galactosidase activities ' (1994 ) 60 Appl. Environ. Microbiol. : 12 -28.

    • Search Google Scholar
  • Naár, Z., Kecskés, M. (1998) Factors influencing the competitive saprophytic ability of Trichoderma species. Microbiol. Res. 153 , 119–129.

    Kecskés M. , 'Factors influencing the competitive saprophytic ability of Trichoderma species ' (1998 ) 153 Microbiol. Res. : 119 -129.

    • Search Google Scholar
  • Payne, D., Gregory, P. J. (1988) The temperature of the soil. In: Wild, E. (ed.) Russel’s Soil Conditions and Plant Growth , 11th edn. Longman Scientific and Technical, Harlow, U.K., pp. 282–297.

    Gregory P. J. , '', in Russel’s Soil Conditions and Plant Growth , (1988 ) -.

  • Rifai, M. A. (1969) A revision of the genus Trichoderma . Mycol. Pap. 116 , 1–56.

    Rifai M. A. , 'A revision of the genus Trichoderma ' (1969 ) 116 Mycol. Pap. : 1 -56.

  • Rigó, K., Varga, J., Tóth, B., Mesterházy, Á., Kozakiewicz, Z. (2002) Phylogenetic analysis of Aspergillus section Flavi based on sequences of the internal transcribed spacer regions and the 5.8 S rRNA gene. J. Gen. Appl. Microbiol. 48 , 9–16.

    Kozakiewicz Z. , 'Phylogenetic analysis of Aspergillus section Flavi based on sequences of the internal transcribed spacer regions and the 5.8 S rRNA gene ' (2002 ) 48 J. Gen. Appl. Microbiol. : 9 -16.

    • Search Google Scholar
  • Schmoll, M., Kubicek, C. P. (2003) Regulation of Trichoderma cellulase formation: lessons in molecular biology from an industrial fungus. Acta Microbiol. Immunol. Hung. 50 , 125–140.

    Kubicek C. P. , 'Regulation of Trichoderma cellulase formation: lessons in molecular biology from an industrial fungus ' (2003 ) 50 Acta Microbiol. Immunol. Hung. : 125 -140.

    • Search Google Scholar
  • Suarez, B., Rey, M., Castillo, P., Monte, E., Llobell, A. (2004) Isolation and characterization of PRA1, a trypsin-like protease from the biocontrol agent Trichoderma harzianum CECT 2413 displaying nematicidal activity. Appl. Microbiol. Biotechnol. 65 , 46–55.

    Llobell A. , 'Isolation and characterization of PRA1, a trypsin-like protease from the biocontrol agent Trichoderma harzianum CECT 2413 displaying nematicidal activity ' (2004 ) 65 Appl. Microbiol. Biotechnol. : 46 -55.

    • Search Google Scholar
  • Szekeres, A., Kredics, L., Antal, Z., Manczinger, L., Kevei, F. (2004) Isolation and characterization of protease overproducing mutants of Trichoderma harzianum . FEMS Microbiol. Lett. 233 , 215–222.

    Kevei F. , 'Isolation and characterization of protease overproducing mutants of Trichoderma harzianum ' (2004 ) 233 FEMS Microbiol. Lett. : 215 -222.

    • Search Google Scholar
  • Szekeres, A., Láday, M., Kredics, L., Antal, Z., Hatvani, L., Manczinger, L. (2004) Investigation of Trichoderma strains isolated from winter wheat rhizosphere. IOBC/WPRS Bull. 27 , 155–158.

    Manczinger L. , 'Investigation of Trichoderma strains isolated from winter wheat rhizosphere ' (2004 ) 27 IOBC/WPRS Bull. : 155 -158.

    • Search Google Scholar
  • Uchikoba, T., Mase, T., Arima, K., Yonezawa, H., Kaneda, M. (2001) Isolation and characterization of a trypsin-like protease from Trichoderma viride . Biol. Chem. 382 , 1509–1513.

    Kaneda M. , 'Isolation and characterization of a trypsin-like protease from Trichoderma viride ' (2001 ) 382 Biol. Chem. : 1509 -1513.

    • Search Google Scholar
  • Vazquez-Garciduenas, S., Leal-Morales, C. A., Herrera-Estrella, A. (1998) Analysis of the β-1,3-glucanolytic system of the biocontrol agent Trichoderma harzianum . Appl. Environ. Microbiol. 64 , 1442–1446.

    Herrera-Estrella A. , 'Analysis of the β-1,3-glucanolytic system of the biocontrol agent Trichoderma harzianum ' (1998 ) 64 Appl. Environ. Microbiol. : 1442 -1446.

    • Search Google Scholar