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A novel strain of Brevibacillus laterosporus produces chitinases that contribute to its biocontrol potential

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Abstract

A novel strain exhibiting entomopathogenic and chitinolytic activity was isolated from mangrove marsh soil in India. The isolate was identified as Brevibacillus laterosporus by phenotypic characterization and 16S rRNA sequencing and designated Lak1210. When grown in the presence of colloidal chitin as the sole carbon source, the isolate produced extracellular chitinases. Chitinase activity was inhibited by allosamidin indicating that the enzymes belong to the family 18 chitinases. The chitinases were purified by ammonium sulfate precipitation followed by chitin affinity chromatography yielding chitinases and chitinase fragments with 90, 75, 70, 55, 45, and 25 kDa masses. Mass spectrometric analyses of tryptic fragments showed that these fragments belong to two distinct chitinases that are almost identical to two putative chitinases, a 89.6-kDa four-domain chitodextrinase and a 69.4-kDa two-domain enzyme called ChiA1, that are encoded on the recently sequenced genome of B. laterosporus LMG15441. The chitinase mixture showed two pH optima, at 6.0 and 8.0, and an optimum temperature of 70 °C. The enzymes exhibited antifungal activity against the phytopathogenic fungus Fusarium equiseti. Insect toxicity bioassays with larvae of diamondback moths (Plutella xylostella), showed that addition of chitinases reduced the time to reach 50 % mortality upon infection with non-induced B. laterosporus from 3.3 to 2.1 days. This study provides evidence for the presence of inducible, extracellular chitinolytic enzymes in B. laterosporus that contribute to the strain’s antifungal activity and insecticidal activity.

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Acknowledgments

This work was funded by grant 178428 from the Norwegian Research Council. We thank Morten Skaugen and Magnus Arntzen at The NorProteomics Consortium for valuable help with the protein identification, Anne Cathrine Bunæs for technical assistance and Torfinn Torp at the Norwegian Institute for Agricultural and Environmental Research for help with the statistical analysis.

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Authors and Affiliations

  1. Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, P.O. Box 5003, 1432, Ås, Norway

    Lakshmi Prasanna, Vincent G. H. Eijsink & Sigrid Gåseidnes

  2. Norwegian Institute for Agricultural and Environmental Research, Plant Health and Plant Protection Division, Høgskoleveien 7, 1432, Ås, Norway

    Richard Meadow

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  1. Lakshmi Prasanna
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  2. Vincent G. H. Eijsink
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  3. Richard Meadow
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  4. Sigrid Gåseidnes
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Correspondence to Vincent G. H. Eijsink.

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Prasanna, L., Eijsink, V.G.H., Meadow, R. et al. A novel strain of Brevibacillus laterosporus produces chitinases that contribute to its biocontrol potential. Appl Microbiol Biotechnol 97, 1601–1611 (2013). https://doi.org/10.1007/s00253-012-4019-y

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  • DOI: https://doi.org/10.1007/s00253-012-4019-y

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