The endochitinase ChiA Btt of Bacillus thuringiensis subsp. tenebrionis DSM‐2803 and its potential use to control the phytopathogen Colletotrichum gloeosporioides

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 E...

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Bibliographic Details
Published in:	MicrobiologyOpen (Weinheim) Vol. 5; no. 5; pp. 819 - 829
Main Authors:	Fuente‐Salcido, Norma M., Casados‐Vázquez, Luz E., García‐Pérez, Ada P., Barboza‐Pérez, Uriel E., Bideshi, Dennis K., Salcedo‐Hernández, Rubén, García‐ Almendarez, Blanca E., Barboza‐Corona, José E.
Format:	Journal Article
Language:	English
Published:	England John Wiley & Sons, Inc 01-10-2016 John Wiley and Sons Inc
Subjects:	Anthracnose Antifungal Agents - metabolism Antifungal Agents - pharmacology Bacillus thuringiensis Bacillus thuringiensis - enzymology Bacillus thuringiensis subsp. tenebrionis Biological Control Agents Cations ChiA Btt Chitinase Chitinases - genetics Chitinases - metabolism Chitinases - pharmacology Cloning Colletotrichium gloeosporioides Colletotrichum - drug effects Colletotrichum gloeosporioides Divalent cations DSM‐2803 E coli Endochitinase Enzymes Escherichia coli Escherichia coli - genetics Escherichia coli - metabolism Etiology Fungi Gene amplification Histidine Mercury Metabolites Original Research Pest control Pests phytopathogen Plant Diseases - microbiology Plasmids Recombinant Proteins - metabolism Recombinant Proteins - pharmacology ChiA Btt Bacillus thuringiensis subsp. tenebrionis DSM-2803 endochitinase phytopathogen Colletotrichium gloeosporioides
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Summary:	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. It is reported for the first time the cloning and characterization of a chitinase synthesized by the commercial strain of Bacillus thuringiensis subsp. tenebrionis DSM‐2803.We show that purified recombinant enzyme from B. thuringiensis is active against the fungus Colletotrichium gloeosporioides.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 N.M de la F‐S. and L.E.C‐V. contributed equally to this article.
ISSN:	2045-8827 2045-8827
DOI:	10.1002/mbo3.372