Biochemical characterization of a 27 kDa 1,3-β-d-glucanase from Trichoderma asperellum induced by cell wall of Rhizoctonia solani |
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Authors: | Raquel da Silva AiresAndrei Stecca Steindorff Marcelo Henrique Soller RamadaSaulo José Linhares de Siqueira Cirano José Ulhoa |
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Institution: | a Instituto de Ensino Superior de Porto Nacional, 77500-000, Porto Nacional, TO, Brazil b Laboratório de Enzimologia, Instituto de Ciências Biológicas II, Universidade Federal de Goiás, 74.690-900 Goiânia, GO, Brazil |
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Abstract: | Trichoderma asperellum produces two extracellular 1,3-β-d-glucanase upon induction with cell walls from Rhizoctonia solani. A minor 1,3-β-d-glucanase was purified to homogeneity by ion exchange chromatography on Q-Sepharose and gel filtration on Sephacryl S-100. A typical procedure provided 13.8-fold purification with 70% yield. SDS-PAGE of the purified enzyme showed a single protein band of molecular weight 27 kDa. The enzyme exhibited optimum catalytic activity at pH 3.6 and 45 °C. It was thermostable at 40 °C, and retained 75% activity after 60 min at 45 °C. The Km and Vmax values for 1,3-β-d-glucanase, using laminarin as substrate, were 0.323 mg ml−1 and 0.315 U min−1, respectively. The enzyme was strongly inhibited by Hg2+ and SDS. The enzyme was only active toward glucans containing β-1,3-linkages. Peptide sequences showed similarity with two endo-1,3(4)-β-d-glucanases from Aspergillus fumigatus Af293when compared against GenBank non-redundant database. |
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Keywords: | Trichoderma asperellum 1 d-glucanase" target="_blank">3-β-d-glucanase Purification Characterization |
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