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2022 Volume 13 Issue 2

Production and Partial Purification of Thermostable Chitinase from Bacillus cereus BSH-4 and its Antifungal Activity


Basavaraj Shrishailappa Hungund, Gururaj Basavaraj Tennalli, Sharanappa Achappa, Manisha Nagaraj Divate
Abstract

The study highlights the production of chitinase from Bacillus cereus BSH-4, which was isolated from a marine soil source and identified by 16S rRNA gene sequencing. After 96 hours of incubation in shake flask conditions profiling studies showed maximum enzyme activity and growth. The ammonium sulfate precipitation method was used at 80% saturation for partial enzyme purification with further purification done by dialysis wherein the purification fold increased to 1.75. The sodium dodecyl sulfate gel electrophoresis (SDS-PAGE) and silver staining methods were employed to assess the enzyme molecular weight observed to be 60 kDa. The enzyme activity and stability process parameters effects like temperature and pH were studied. At a pH value of 7.0, the greatest enzyme activity was noted which was relatively retained at 80% till pH 9.0. Similarly, higher enzyme activity was observed at a temperature of 30oC and retained 70% relative activity till 60oC, indicating the thermostable nature of the enzyme. Studies on the influence of metal ions showed that Zn2+, Cu2+, Ca2+, Mg2+, Mn2+, and Fe2+ inhibited the enzyme by 20% to 80%. An investigation was also done on the influence of organic solvents like methanol, ethanol, acetone, and isopropyl alcohol on enzyme stability. The results showed that the enzyme was stable in all the solvents, exhibiting 43% to 70% relative activity. The enzyme kinetic studies indicated Km and Vmax values of 4.54 mg ml-1 and 76.92 UmL-1, respectively. The enzyme also demonstrated antifungal activity against Aspergillus niger NCIM 1207 using the agar well diffusion method.


How to cite this article
Vancouver
Hungund BS, Tennalli GB, Achappa S, Divate MN. Production and Partial Purification of Thermostable Chitinase from Bacillus cereus BSH-4 and its Antifungal Activity. J Biochem Technol. 2022;13(2):46-53. https://doi.org/10.51847/RAlF1H5BCe
APA
Hungund, B. S., Tennalli, G. B., Achappa, S., & Divate, M. N. (2022). Production and Partial Purification of Thermostable Chitinase from Bacillus cereus BSH-4 and its Antifungal Activity. Journal Of Biochemical Technology, 13(2),46-53. https://doi.org/10.51847/RAlF1H5BCe
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JOURNAL OF BIOCHEMICAL TECHNOLOGY
JOURNAL OF BIOCHEMICAL TECHNOLOGY
Journal of Biochemical Technology is a peer reviewed International Journal published by the Sevas Publishing on behalf of the Biochemical Technology Society, a Registered Charity Organization from India

AREA OF INTEREST
AREA OF INTEREST
Areas of high interest, cover new advances in enzymatic and protein mechanims; applied molecular genetics and biotechnology; genomics and proteomics; metabolic; medical, environmental, food and agro biotechnology.

FOCUS AND SCOPE
FOCUS AND SCOPE
Journal Of Biochemical Technology” Provides A Medium For The Rapid Publication Of Full-Length Articles, Mini-Reviews Of New And Emerging Products And Short Communications On All Aspects Of ...

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This journal provides immediate open access to its content on the principle that making research freely available to the public supports a greater global exchange of knowledge. Keywords include, Biochemical Research: Endo/exocytosis, Trafficking, Membrane Biology, Cell Migration, Cell-Matrix Organelle Biogenesis, Cytoskeleton Proteolysis, Cell Death, Cell Cycle, Cancer, Cell Growth/Death, Differentiation, Drug Targets, Gene Therapy, Models of Disease, Proteomics, Stem Cells, Bioenergetics, Mitochondria, Free Radicals, Redox Signaling, Ion Transport/Channels, Oxidative