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Chitin-degrading enzymes from an actinomycete ectosymbiont of Acromyrmex subterraneus brunneus (Hymenoptera: Formicidae)

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Abstract

Microbes have many mechanisms to exert their inhibitory activity against target pests. One such mechanism involves the production and secretion of hydrolytic enzymes, such as chitinases, which are produced naturally by plants in response to attack by insect herbivores and phytopathogens and have been sought as an additional factor to enhance pest management. Thus, our main aim was to screen the diverse actinomycete community associated with the integument of Acromyrmex subterraneus brunneus for a chitinase-producing strain and to characterize its chitinases. We identified isolate ENT-21—a Streptomyces sp.—as a chitinase-producer and our data indicate that this isolate produces a chitinolytic complex that contains a chitinase and a high-molecular-weight β-N-acetylglucosaminidase (>100 kDa) when cultured in Chitin-Czapek broth. The presence of chitinases in the genome of this isolate was checked by diagnostic PCR, and two chitinase genes belonging to family 18 group A and family 19 were verified. The chitinolytic activity of the crude extract was observed at pH values ranging from 3.8 to 11.0, with the highest chitinase activities recorded at pH 9.0 and 9.5, whereas optimum β-N-acetylglucosaminidase activity was observed over a narrow pH range, between pH 4.7 and 5.1. We describe some biochemical and molecular properties of the chitinase and β-N-acetylglucosaminidase produced by ENT-21, and discuss the potential for exploitation of these enzymes for pest control.

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Acknowledgments

The authors appreciate the financial support provided by FAPESP to G.D.R. (PhD fellowship—process 2012/50021-1), T.D.Z. (PostDoctoral fellowship—processes 2007/58712-5 and 2011/14333-6), A.S.G. (PhD fellowship—process 2012/04287-0) and A.P. (Masters fellowship—process 2010/13675-8). F.L.C. also acknowledges FAPESP for funding this research (grants—processes 2007/59019-1; 2011/50877-0).

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Authors and Affiliations

  1. Lab de Interações em Insetos, Depto de Entomologia & Acarologia, Escola Superior de Agricultura “Luiz de Queiroz”, Univ de São Paulo (ESALQ/USP), Piracicaba, SP, Brazil

    Guilherme Duarte Rossi, Tiago Domingues Zucchi, Aline Sartori Guidolin, Aline Peruchi & Fernando Luis Cônsoli

  2. Depto de Fitossanidade, Faculdade de Ciências Agrárias e Veterinárias, FCAV/UNESP, Jaboticabal, SP, Brazil

    Guilherme Duarte Rossi

  3. Lab Microbiologia Ambiental, EMBRAPA Meio Ambiente, Jaguariúna, SP, Brazil

    Tiago Domingues Zucchi

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  1. Guilherme Duarte Rossi
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  2. Tiago Domingues Zucchi
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Correspondence to Guilherme Duarte Rossi or Fernando Luis Cônsoli.

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Rossi, G.D., Zucchi, T.D., Guidolin, A.S. et al. Chitin-degrading enzymes from an actinomycete ectosymbiont of Acromyrmex subterraneus brunneus (Hymenoptera: Formicidae). Ann Microbiol 65, 565–574 (2015). https://doi.org/10.1007/s13213-014-0892-1

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  • DOI: https://doi.org/10.1007/s13213-014-0892-1

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