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Production, Purification, and Characterization of an Industrially Important Enzyme Alkaline Protease Produced from Locally Isolated Bacillus Bacteria

Received: 9 November 2021     Accepted: 9 December 2021     Published: 24 December 2021
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Abstract

The worldwide demand for industrially essential enzymes is growing due to their eco-friendly and sustainable applications in broad industrial sectors. The main objective of this investigation is intended to isolate potent alkalophilic native bacteria from local habitats for the cost-effective production of an enzyme alkaline protease. For screening of alkalophilic bacteria, bacterial samples were taken from various natural alkaline habitats, and bacterial cultures were screened on agar plate having skimmed milk as protein substrate by using the protein hydrolysis method. The bacteria showing maximum proteolytic potential were identified by microscopic, biochemical, and 16S rDNA analysis. Additionally, the culture components and other medium parameters have been optimized for higher enzyme production. High yield alkaline protease was obtained with conditions of 1% inoculum, pH 9.0, and at 37°C temperature, with 1% sugarcane molasses as the best suitable carbon substrate. Partial pur¬ification of the enzyme was performed and characterized for their optimum activity on various parameters (temperatures and pH range). The approx. molecular weight of the enzyme was estimated to be ~35 kDa. This study demonstrates the production of an industrially important enzyme (alkaline protease) from a newly isolated native Bacillus spp. Economical production of this enzyme can be commercially applied in vast industrial sectors like agriculture, textile, food, detergent, and leather industries.

Published in International Journal of Microbiology and Biotechnology (Volume 6, Issue 4)
DOI 10.11648/j.ijmb.20210604.15
Page(s) 137-146
Creative Commons

This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.

Copyright

Copyright © The Author(s), 2021. Published by Science Publishing Group

Keywords

Agro-industrial Residues, Alkaline Protease, Bacillus Spp, Cost-effective, Sugarcane Molasses

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    Ranjeet Singh, Ravindra Meena, Harsh Kumar, Banwari Lal. (2021). Production, Purification, and Characterization of an Industrially Important Enzyme Alkaline Protease Produced from Locally Isolated Bacillus Bacteria. International Journal of Microbiology and Biotechnology, 6(4), 137-146. https://doi.org/10.11648/j.ijmb.20210604.15

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    ACS Style

    Ranjeet Singh; Ravindra Meena; Harsh Kumar; Banwari Lal. Production, Purification, and Characterization of an Industrially Important Enzyme Alkaline Protease Produced from Locally Isolated Bacillus Bacteria. Int. J. Microbiol. Biotechnol. 2021, 6(4), 137-146. doi: 10.11648/j.ijmb.20210604.15

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    AMA Style

    Ranjeet Singh, Ravindra Meena, Harsh Kumar, Banwari Lal. Production, Purification, and Characterization of an Industrially Important Enzyme Alkaline Protease Produced from Locally Isolated Bacillus Bacteria. Int J Microbiol Biotechnol. 2021;6(4):137-146. doi: 10.11648/j.ijmb.20210604.15

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  • @article{10.11648/j.ijmb.20210604.15,
      author = {Ranjeet Singh and Ravindra Meena and Harsh Kumar and Banwari Lal},
      title = {Production, Purification, and Characterization of an Industrially Important Enzyme Alkaline Protease Produced from Locally Isolated Bacillus Bacteria},
      journal = {International Journal of Microbiology and Biotechnology},
      volume = {6},
      number = {4},
      pages = {137-146},
      doi = {10.11648/j.ijmb.20210604.15},
      url = {https://doi.org/10.11648/j.ijmb.20210604.15},
      eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ijmb.20210604.15},
      abstract = {The worldwide demand for industrially essential enzymes is growing due to their eco-friendly and sustainable applications in broad industrial sectors. The main objective of this investigation is intended to isolate potent alkalophilic native bacteria from local habitats for the cost-effective production of an enzyme alkaline protease. For screening of alkalophilic bacteria, bacterial samples were taken from various natural alkaline habitats, and bacterial cultures were screened on agar plate having skimmed milk as protein substrate by using the protein hydrolysis method. The bacteria showing maximum proteolytic potential were identified by microscopic, biochemical, and 16S rDNA analysis. Additionally, the culture components and other medium parameters have been optimized for higher enzyme production. High yield alkaline protease was obtained with conditions of 1% inoculum, pH 9.0, and at 37°C temperature, with 1% sugarcane molasses as the best suitable carbon substrate. Partial pur¬ification of the enzyme was performed and characterized for their optimum activity on various parameters (temperatures and pH range). The approx. molecular weight of the enzyme was estimated to be ~35 kDa. This study demonstrates the production of an industrially important enzyme (alkaline protease) from a newly isolated native Bacillus spp. Economical production of this enzyme can be commercially applied in vast industrial sectors like agriculture, textile, food, detergent, and leather industries.},
     year = {2021}
    }
    

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  • TY  - JOUR
    T1  - Production, Purification, and Characterization of an Industrially Important Enzyme Alkaline Protease Produced from Locally Isolated Bacillus Bacteria
    AU  - Ranjeet Singh
    AU  - Ravindra Meena
    AU  - Harsh Kumar
    AU  - Banwari Lal
    Y1  - 2021/12/24
    PY  - 2021
    N1  - https://doi.org/10.11648/j.ijmb.20210604.15
    DO  - 10.11648/j.ijmb.20210604.15
    T2  - International Journal of Microbiology and Biotechnology
    JF  - International Journal of Microbiology and Biotechnology
    JO  - International Journal of Microbiology and Biotechnology
    SP  - 137
    EP  - 146
    PB  - Science Publishing Group
    SN  - 2578-9686
    UR  - https://doi.org/10.11648/j.ijmb.20210604.15
    AB  - The worldwide demand for industrially essential enzymes is growing due to their eco-friendly and sustainable applications in broad industrial sectors. The main objective of this investigation is intended to isolate potent alkalophilic native bacteria from local habitats for the cost-effective production of an enzyme alkaline protease. For screening of alkalophilic bacteria, bacterial samples were taken from various natural alkaline habitats, and bacterial cultures were screened on agar plate having skimmed milk as protein substrate by using the protein hydrolysis method. The bacteria showing maximum proteolytic potential were identified by microscopic, biochemical, and 16S rDNA analysis. Additionally, the culture components and other medium parameters have been optimized for higher enzyme production. High yield alkaline protease was obtained with conditions of 1% inoculum, pH 9.0, and at 37°C temperature, with 1% sugarcane molasses as the best suitable carbon substrate. Partial pur¬ification of the enzyme was performed and characterized for their optimum activity on various parameters (temperatures and pH range). The approx. molecular weight of the enzyme was estimated to be ~35 kDa. This study demonstrates the production of an industrially important enzyme (alkaline protease) from a newly isolated native Bacillus spp. Economical production of this enzyme can be commercially applied in vast industrial sectors like agriculture, textile, food, detergent, and leather industries.
    VL  - 6
    IS  - 4
    ER  - 

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Author Information
  • Environmental and Industrial Biotechnology Division, The Energy and Resources Institute, New Delhi, India

  • Environmental and Industrial Biotechnology Division, The Energy and Resources Institute, New Delhi, India

  • Environmental and Industrial Biotechnology Division, The Energy and Resources Institute, New Delhi, India

  • Environmental and Industrial Biotechnology Division, The Energy and Resources Institute, New Delhi, India

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