Isolation and characterization of PRA1, a trypsin-like protease from the biocontrol agent Trichoderma harzianum CECT 2413 displaying nematicidal activity

Applied Microbiology and Biotechnology
Belen SuarezA Llobell

Abstract

Mycoparasitic Trichoderma strains secrete a complex set of hydrolytic enzymes under conditions related to antagonism. Several proteins with proteolytic activity were detected in culture filtrates from T. harzianum CECT 2413 grown in fungal cell walls or chitin and the protein responsible for the main activity (PRA1) was purified to homogeneity. The enzyme was monomeric, its estimated molecular mass was 28 kDa (SDS-PAGE), and its isoelectric point 4.7-4.9. The substrate specificity and inhibition profile of the enzyme correspond to a serine-protease with trypsin activity. Synthetic oligonucleotide primers based on N-terminal and internal sequences of the protein were designed to clone a full cDNA corresponding to PRA1. The protein sequence showed <43% identity to mammal trypsins and 47-57% to other fungal trypsin-like proteins described thus far. Northern analysis indicated that PRA1 is induced by conditions simulating antagonism, is subject to nitrogen and carbon derepression, and is affected by pH in the culture media. The number of hatched eggs of the root-knot nematode Meloidogyne incognita was significantly reduced after incubation with pure PRA1 preparations. This nematicidal effect was improved using fungal culture filtra...Continue Reading

Citations

Jul 9, 2013·Applied Microbiology and Biotechnology·Jinkui YangKe-Qin Zhang
May 17, 2013·World Journal of Microbiology & Biotechnology·Xiaoxue YangJunzheng Zhang
May 3, 2011·Applied and Environmental Microbiology·Barbara ReithnerAlfredo Herrera-Estrella
Jul 29, 2006·BMC Genomics·Juan Antonio VizcaínoManuel Rey
Mar 29, 2011·Journal of Environmental Management·Petri SusiTimo Korpela
Jun 20, 2007·Letters in Applied Microbiology·J L PereiraO L Franco
Jun 30, 2015·Environmental Microbiology·Jian ZhangIrina S Druzhinina
Apr 6, 2004·FEMS Microbiology Letters·András SzekeresLászló Manczinger
Sep 21, 2007·Fungal Genetics and Biology : FG & B·Miguel A Moreno-MateosTahía Benítez
Sep 26, 2013·Journal of Molecular Microbiology and Biotechnology·Yan Liu, Qian Yang
Jul 12, 2005·Acta Microbiologica Et Immunologica Hungarica·L KredicsErzsébet Nagy
Sep 17, 2011·Nature Reviews. Microbiology·Irina S DruzhininaChristian P Kubicek
Mar 3, 2007·Applied Microbiology and Biotechnology·Juan Antonio VizcaínoEnrique Monte

❮ Previous
Next ❯