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Purification and characterization of antifungal chitinase from Bacillus safensis MBCU6 and its application for production of chito-oligosaccharides

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Abstract

The chitinase producing bacterial strain was isolated from the vermicompost amended site of Mehsana district of Gujarat, India, and identified as Bacillus safensis MBCU6 using 16S rDNA sequencing. The chitinase was purified by ammonium sulfate precipitation followed by diethylaminoethanol sepharose CL-6B column chromatography. The purified enzyme could be demonstrated as a single band on sodium dodecyl sulfate polyacryalamide gel electrophoresis analysis as well as clear zone on zymogram, with estimated molecular mass of 58 kDa. The optimum pH and temperature of chitinase were pH 7.0 and 60°C, respectively. The purified chitinase exhibited high degree of antifungal activity particularly against pathogenic Macrophomina phaseolina (60%) and Rhizoctonia solani (73%) by dissolving their cell wall components. The purified enzyme could hydrolyze colloidal chitin to its oligomers. It infers that the chitinase produced by Bacillus safensis may play a significant role in the activity as a biopesticide and bioactive material production.

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Abbreviations

DEAE:

diethylaminoethanol

NAG:

N-aetylglucosamine

SDS-PAGE:

sodium dodecyl sulfate poly-acryalamide gel electrophoresis

TLC:

thin-layer chromatography

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Acknowledgements

The financial support from Department of Science and Technology (DST), New Delhi, India, under Women Scientist Scheme (WOS-A) (Grant No. SR/WOS-A/LS-186/2010) is gratefully acknowledged. The authors thank Dr. Shailesh R. Waghmare for proof reading of the manuscript.

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  1. Department of Microbiology and Biotechnology, University School of Sciences, Gujarat University, Ahmedabad, 380013, Gujarat, India

    Urja Pandya & Meenu Saraf

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  1. Urja Pandya
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  2. Meenu Saraf
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Correspondence to Meenu Saraf.

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Pandya, U., Saraf, M. Purification and characterization of antifungal chitinase from Bacillus safensis MBCU6 and its application for production of chito-oligosaccharides. Biologia 70, 863–868 (2015). https://doi.org/10.1515/biolog-2015-0112

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  • DOI: https://doi.org/10.1515/biolog-2015-0112

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