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Production and Biochemical Characterization of Insecticidal Enzymes from Aspergillus fumigatus Toward Callosobruchus maculatus

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Abstract

In the present work, Aspergillus fumigatus is described as a higher producer of hydrolytic enzymes secreted in response to the presence of the Callosobruchus maculatus bruchid pest. This fungus was able to grow over cowpea weevil shells as a unique carbon source, secreting alkaline proteolytic and chitinolytic enzymes. Enzyme secretion in A. fumigatus was induced by both C. maculatus exoskeleton as well as commercial chitin, and alkaline proteolytic and chitinolytic activities were detected after 48 hours of growth. Furthermore, sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) analysis showed the production of specific proteins. Among them, two extracellular alkaline proteinases from culture enriched with C. maculatus exoskeleton were purified after chromatographic procedures using ion exchange and affinity columns. These proteins, named AP15 and AP30, had apparent molecular masses of 15,500 and 30,000 Da, respectively, as estimated by SDS-PAGE electrophoresis and mass spectrometry. AP30 was classified as a serine proteinase because it was inhibited by 5 mM phenylmethylsulfonyl fluoride (100%) and 50 μM leupeptin (67.94%).

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Acknowledgments

This work was supported by a grant research from the Universidade Católica de Brasília. J. Pereira acknowledges C. Bloch Jr. for spectrometric analysis.

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  1. Centro de Análises Proteômicas e Bioquímicas. Programa de Pós-Graduação em Ciências Genômicas e Biotecnologia, Universidade Católica de Brasília, Campus II, 70790-160, Distrito Federal, Brazil

    Jackeline L. Pereira, Octávio L. Franco & Eliane F. Noronha

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  1. Jackeline L. Pereira
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  2. Octávio L. Franco
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  3. Eliane F. Noronha
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Correspondence to Eliane F. Noronha.

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Pereira, J.L., Franco, O.L. & Noronha, E.F. Production and Biochemical Characterization of Insecticidal Enzymes from Aspergillus fumigatus Toward Callosobruchus maculatus . Curr Microbiol 52, 430–434 (2006). https://doi.org/10.1007/s00284-005-0192-x

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