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Purification and Characterization of Alkaline Protease from Bacillus sp. HD292

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Proceedings of the National Academy of Sciences, India Section B: Biological Sciences Aims and scope Submit manuscript

Abstract

Proteolytic enzymes form a very significant group of enzymes which are used for a variety of purposes including as a detergent additive. In the present investigation, the protein hydrolyzing enzyme secreted by Bacillus sp. HD292 was purified and characterized. The purification of the proteolytic enzyme was undertaken by ammonium sulphate fractionation (20–50%) and gel filtration chromatography using Sephadex G-75 which resulted in 17.78 fold purification. On performing SDS-PAGE analysis, a single band of approx. 30 kD was observed. The purified protease displayed peak activity at pH 9.5 and 70 °C. The Km value of the enzyme was found to be 2.7 mg ml−1 with casein as the substrate. pBLAST analysis of the translated sequence resulting from the PCR product obtained from the conserved region of the gene, showed it to be similar to various intracellular and extracellular serine proteases. Also, the purified bacterial protease was found to retain its proteolytic activity at high pH as well as high temperature and was also observed to be quite compatible with the detergent Paras (R3 Organics).

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Acknowledgements

The authors express gratitude to Council of Scientific & Industrial Research, India for financial support in the form of JRF and SRF (F. NO. 10-2(5)/2005(ii)- E.U.II).

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Author notes

  1. Chayanika Putatunda

    Present address: Department of Microbiology, DAV University, Jalandhar, Punjab, India

Authors and Affiliations

  1. Department of Microbiology, C.C.S. Haryana Agricultural University, Hisar, India

    Chayanika Putatunda, B. S. Kundu & Ranjana Bhatia

Authors
  1. Chayanika Putatunda
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  2. B. S. Kundu
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  3. Ranjana Bhatia
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Correspondence to Chayanika Putatunda.

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Significance statement

Alkaline proteases are used in a host of industries. Bacteria are significant producer of such enzymes. So, the present investigation was done to assess whether the protease produced by Bacillus sp. HD292 may be used as a detergent additive.

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Putatunda, C., Kundu, B.S. & Bhatia, R. Purification and Characterization of Alkaline Protease from Bacillus sp. HD292. Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci. 89, 957–965 (2019). https://doi.org/10.1007/s40011-018-1011-z

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  • DOI: https://doi.org/10.1007/s40011-018-1011-z

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