The endochitinase ChiA Btt ofBacillus thuringiensissubsp.tenebrionisDSM ‐2803 and its potential use to control the phytopathogenColletotrichum gloeosporioides
Autor: | Uriel E. Barboza-Perez, José E. Barboza-Corona, Dennis K. Bideshi, Norma M de la Fuente-Salcido, Ada P. García‐Pérez, Rubén Salcedo-Hernández, Luz E. Casados-Vázquez, Blanca E. García Almendárez |
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Rok vydání: | 2016 |
Předmět: |
0106 biological sciences
0301 basic medicine Antifungal Agents Endochitinase activity Bacillus thuringiensis medicine.disease_cause 01 natural sciences Microbiology Bacillus thuringiensis subsp. tenebrionis ChiA Btt phytopathogen 03 medical and health sciences Affinity chromatography 010608 biotechnology Colletotrichium gloeosporioides Colletotrichum Escherichia coli medicine Original Research Plant Diseases biology Chitinases biology.organism_classification Recombinant Proteins Enzyme assay Spore DSM‐2803 030104 developmental biology Biological Control Agents Chitinase biology.protein endochitinase |
Zdroj: | MicrobiologyOpen |
ISSN: | 2045-8827 |
DOI: | 10.1002/mbo3.372 |
Popis: | Bacillus thuringiensis subsp. tenebrionis DSM‐2803 has been studied extensively and spore/crystal mixtures of this strain are used widely in commercial products to control coleopteran pests. The endochitinase chiA Btt gene of B. thuringiensis subsp. tenebrionis DSM‐2803 was cloned and expressed in Escherichia coli. The recombinant 6x‐histidine tagged protein (rChiA Btt, ~74 kDa), was purified by a HiTrap Ni affinity column. The Km of rChiA Btt was 0.847 μmol L−1 and its optimal activity occurred at pH 7 and ~40°C. Most divalent cations reduced endochitinase activity but only Hg+2 abolished activity of the enzyme. We report for the first time the characterization of a chitinase synthesized by B. thuringiensis subsp. tenebrionis DSM‐2803, and show that the purified rChiA74 Btt reduced the radial growth and increased the hyphal density of Colletotrichium gloeosporioides, the etiological agent of “anthracnose” in plants. |
Databáze: | OpenAIRE |
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