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Extraction, Purification and Characterization of Protease from Aspergillus Niger Isolated from Yam Peels

Received: 30 October 2014     Accepted: 12 November 2014     Published: 16 February 2015
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Abstract

Protease was obtained from Aspergillus niger isolated from yam peels; a food waste, purified and characterized. Purification was achieved using ion exchange DEAE column and gel filtration (Sephadex G-200) chromatography. Effects of temperature; pH and production time on protease production were investigated. Also, physicochemical characteristics of the purified enzyme were investigated. The optimum production of protease was at temperature, pH and time of 37oC, 7.0 and 42 hrs respectively. The results showed that purified protease had more specific enzymatic activity than crude samples from Aspergillus Niger. whereas the specific activity of crude enzyme was 0.51 (U/mg), while the purified enzyme had an improved specific activity of 8.51 (U/mg).Optimum temperature and pH values of the purified protease were found to be 50°C and 10.0, respectively. pH stability of the enzyme ranged from 3.0- 12.0. At 3.0 and 10.0 it retained 70% and 60% of its activity after 5 hrs of incubation. Temperature stability ranged between 30oC and 90oC but most stable at 50oC retaining 94% of its activity after 1 h of incubation. The enzyme exhibited maximum activity on casein, among other protein substrates. EDTA, Cu2+, Fe2+, Mg 2+, and Ca2+ inhibited its activity while Na+ enhanced it. The enzyme was purified 16.60-fold, had a yield of 10.96 and the apparent molecular weight was 46.90 kDa. The study revealed that protease from A. niger can be exploited for protein conversion biotechnologies.

Published in International Journal of Nutrition and Food Sciences (Volume 4, Issue 2)
DOI 10.11648/j.ijnfs.20150402.11
Page(s) 125-131
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), 2015. Published by Science Publishing Group

Keywords

Protease, Aspergillus Niger, Yam Peels, Fermentation, Purification, Characterisation

References
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    Oludumila Omolara Racheal, Abu Temitope Folagbade Ahmed, Enujiugha Victor Ndigwe, Sanni David Morakinyo. (2015). Extraction, Purification and Characterization of Protease from Aspergillus Niger Isolated from Yam Peels. International Journal of Nutrition and Food Sciences, 4(2), 125-131. https://doi.org/10.11648/j.ijnfs.20150402.11

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    ACS Style

    Oludumila Omolara Racheal; Abu Temitope Folagbade Ahmed; Enujiugha Victor Ndigwe; Sanni David Morakinyo. Extraction, Purification and Characterization of Protease from Aspergillus Niger Isolated from Yam Peels. Int. J. Nutr. Food Sci. 2015, 4(2), 125-131. doi: 10.11648/j.ijnfs.20150402.11

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    AMA Style

    Oludumila Omolara Racheal, Abu Temitope Folagbade Ahmed, Enujiugha Victor Ndigwe, Sanni David Morakinyo. Extraction, Purification and Characterization of Protease from Aspergillus Niger Isolated from Yam Peels. Int J Nutr Food Sci. 2015;4(2):125-131. doi: 10.11648/j.ijnfs.20150402.11

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  • @article{10.11648/j.ijnfs.20150402.11,
      author = {Oludumila Omolara Racheal and Abu Temitope Folagbade Ahmed and Enujiugha Victor Ndigwe and Sanni David Morakinyo},
      title = {Extraction, Purification and Characterization of Protease from Aspergillus Niger Isolated from Yam Peels},
      journal = {International Journal of Nutrition and Food Sciences},
      volume = {4},
      number = {2},
      pages = {125-131},
      doi = {10.11648/j.ijnfs.20150402.11},
      url = {https://doi.org/10.11648/j.ijnfs.20150402.11},
      eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ijnfs.20150402.11},
      abstract = {Protease was obtained from Aspergillus niger isolated from yam peels; a food waste, purified and characterized. Purification was achieved using ion exchange DEAE column and gel filtration (Sephadex G-200) chromatography. Effects of temperature; pH and production time on protease production were investigated. Also, physicochemical characteristics of the purified enzyme were investigated. The optimum production of protease was at temperature, pH and time of 37oC, 7.0 and 42 hrs respectively. The results showed that purified protease had more specific enzymatic activity than crude samples from Aspergillus Niger. whereas the specific activity of crude enzyme was 0.51 (U/mg), while the purified enzyme had an improved specific activity of 8.51 (U/mg).Optimum temperature and pH values of the purified protease were found to be 50°C and 10.0, respectively. pH stability of the enzyme ranged from 3.0- 12.0. At 3.0 and 10.0 it retained 70% and 60% of its activity after 5 hrs of incubation. Temperature stability ranged between 30oC and 90oC but most stable at 50oC retaining 94% of its activity after 1 h of incubation. The enzyme exhibited maximum activity on casein, among other protein substrates. EDTA, Cu2+, Fe2+, Mg 2+, and Ca2+ inhibited its activity while Na+ enhanced it. The enzyme was purified 16.60-fold, had a yield of 10.96 and the apparent molecular weight was 46.90 kDa. The study revealed that protease from A. niger can be exploited for protein conversion biotechnologies.},
     year = {2015}
    }
    

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  • TY  - JOUR
    T1  - Extraction, Purification and Characterization of Protease from Aspergillus Niger Isolated from Yam Peels
    AU  - Oludumila Omolara Racheal
    AU  - Abu Temitope Folagbade Ahmed
    AU  - Enujiugha Victor Ndigwe
    AU  - Sanni David Morakinyo
    Y1  - 2015/02/16
    PY  - 2015
    N1  - https://doi.org/10.11648/j.ijnfs.20150402.11
    DO  - 10.11648/j.ijnfs.20150402.11
    T2  - International Journal of Nutrition and Food Sciences
    JF  - International Journal of Nutrition and Food Sciences
    JO  - International Journal of Nutrition and Food Sciences
    SP  - 125
    EP  - 131
    PB  - Science Publishing Group
    SN  - 2327-2716
    UR  - https://doi.org/10.11648/j.ijnfs.20150402.11
    AB  - Protease was obtained from Aspergillus niger isolated from yam peels; a food waste, purified and characterized. Purification was achieved using ion exchange DEAE column and gel filtration (Sephadex G-200) chromatography. Effects of temperature; pH and production time on protease production were investigated. Also, physicochemical characteristics of the purified enzyme were investigated. The optimum production of protease was at temperature, pH and time of 37oC, 7.0 and 42 hrs respectively. The results showed that purified protease had more specific enzymatic activity than crude samples from Aspergillus Niger. whereas the specific activity of crude enzyme was 0.51 (U/mg), while the purified enzyme had an improved specific activity of 8.51 (U/mg).Optimum temperature and pH values of the purified protease were found to be 50°C and 10.0, respectively. pH stability of the enzyme ranged from 3.0- 12.0. At 3.0 and 10.0 it retained 70% and 60% of its activity after 5 hrs of incubation. Temperature stability ranged between 30oC and 90oC but most stable at 50oC retaining 94% of its activity after 1 h of incubation. The enzyme exhibited maximum activity on casein, among other protein substrates. EDTA, Cu2+, Fe2+, Mg 2+, and Ca2+ inhibited its activity while Na+ enhanced it. The enzyme was purified 16.60-fold, had a yield of 10.96 and the apparent molecular weight was 46.90 kDa. The study revealed that protease from A. niger can be exploited for protein conversion biotechnologies.
    VL  - 4
    IS  - 2
    ER  - 

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Author Information
  • Department of Food Science and Technology

  • Department of Food Science and Technology

  • Department of Food Science and Technology

  • Department of Biochemistry

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