American Journal of Biomedical and Life Sciences

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Production Optimization and Characterization of Amylase Enzyme Isolated from Termofil Bacteria Bacillus sp RSAII-1b from Lejja Hot Spring South Sulawesi

Received: 22 November 2015    Accepted: 07 December 2015    Published: 30 December 2015
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Abstract

Thermostable amylase enzyme has a broad commercial value in its use in the processing of starch, sugar production, textile, paper, animal feed, pharmaceuticals and in the manufacture of detergents. This study aims to determine the optimum conditions of amylase production from the termofil bacteria Bacillus sp. RSII-1b isolated from a hot spring Lejja South Sulawesi and characterizing the amylase enzyme. The testing of amylase production was done with various concentration of starch and CaCl2 in the production medium, then fermented to obtain maximum amylase activity, amylase enzyme was produced in optimum condition, and its characteristic was tested using 2% starch substrates in various pH, temperature and determining the compound cofactor which can act as activators or inhibitors of the amylase activity, the enzyme activity was tested using DNS method. Crude extract enzyme has the highest enzyme activity of the protein content determined by the method of Lawry. The results showed that the amylase enzyme from Bacillus sp RSAII-1b isolates can be manufactured to a maximum at 33 hours of fermentation time with conditions: the concentration of substrate (starch) 1.5%, 0.08% CaCl2, 55°C temperature, medium pH 7.0 and aeration speed 200rpm with the activity of 0.1323U/mL, amylase crude extract protein content of 1.86mg/mL, with spesifik activity 0.0711 U/mg protein. Crude extract amylase work optimally at pH 6.0; 55°C -60°C the amylase activity of 0.165U/mL, the specific activity of 0.089U/mg protein. Amylase enzyme is an enzyme that depends on metal because its catalytic activity can be activated by metal ions Ca2+, Mg2+, Cu2+, Ni2+ and Co2+ as activators whereas Zn2+ ions decrease the activity of enzymes as inhibitors. Amylase activity in the crude extract optimum conditions with the addition of 10 mm ions Ca2+ can increase amylase activity up to 32,89%, while the addition of ions Zn2+ can inhibit amylase activity up to 25%.

DOI 10.11648/j.ajbls.20150306.13
Published in American Journal of Biomedical and Life Sciences (Volume 3, Issue 6, December 2015)
Page(s) 115-119
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), 2024. Published by Science Publishing Group

Keywords

Amylase, Hydrolytic, Bacillus sp, Isolate, Starch

References
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[2] Souza, P. M. De., and Magalhães, P. O. De., 2010. Application of Microbial α-amylase in Industry-A review. Braz. J. Microbiol. (Online), Vol 41, No.4 (http://dx.doi.org/10.1590/S1517-83822010000400004, accessed 19 Oktober 2011).
[3] Vieille C and Zeikus G. 2001. Hyperthermophilic Enzymes Source, Uses, and Molecular Mechanism for Thermostability. Microbiology and Molecular Biology Reviews. 65:1–.43.
[4] Vaseekaran, S., Balakumar, S., and Arasaratnam V., 2010, Isolation and Identification of a Bacterial Strain Producing Thermostable α-amylase. Tropical Agricultural Research, 22 (1): 1–11.
[5] Setiasih S., Wahyuntari B., Trimillah, Aprilliani D. 2006. Karakterisasi Enzim α-Amilase Ekstrasel dari Isolat Bakteri Termofil SW2. Jurnal Kimia Indonesia. 1 (1): 22-27.
[6] Rosmimik, Richana, N., Lestari, P., dan Damardjati, Dj. S., 2001. Studi Penambahan Ion Kalsium terdadap Aktifitas dan stabilitas α-amilase Bacillusstearo thermophilus TII12. J. Mikrobiologi Indonesia. 6 (1): 12–14.
[7] Raharjo, S. Ardiansyah, Endang, P. Tien, 2010. Isolation of Thermostable α-amylase from Local Thermophilic Bacteria for Liquefaction. Proceeding ICMNS, 342–354.
[8] Arfah, A. R., Patong R., Ahmad A., Djide N., 2014, Isolasi dan Identifikasi Bakteri Termofil Penghasil Amilase dari Sumber Air Panas Lejja Sulawesi Selatan, Al-kimia Jurnal Penelitian Sains Kimia, 2 (2): 36-46.
[9] Suman S., and Ramesh K., 2010. Production of A Thermostable Extracellular Amylase from Thermophilic Bacillus species. J. Pharm. Sci & Res. 2 (2): 149-154.
[10] Abdel-Fattah,Y. R., Soliman, N. A., El-Toukhy N. M., El-Gendi H., and Ahmed R.S., 2012. Production, Purification and Characterization of Termostable α-amilase Produced by Bacillus licheniformis Isolate AI20. Jurnal Chemistry. 2012: 1–11.
[11] Miller, G. L., 1959; Use of Dinitrosalicylic Acid Reagent for Determination of Reducing Sugar. Anal Chem. 31 (3): 426–428.
[12] Rasooli, I., Astaneh, S. D. A. A., Borna, H., and Barchini K. A., 2008. A Thermostable α-amilase Producing Natural Varient of Bacillus sp. Isolated from Soilin Iran. Am. J. Agri. & Biol. Sci. 3 (3): 591–296.
[13] Lowry, O.H., Rosebrough, N.J., Farr, A.L., and Randall, R.J., 1951, Protein Measurement whit the Folin Phenolreagent. J. Biol. Chem.193:265-275.
[14] Bollag, D. M. and Edelstein, S. J. 1991. Protein Methods. New York : Wiley-Less.
[15] Asad,W., Asif, M., and Rasool, A. S., 2011. Extracellular Enzyme Production by Indigenous thermophilic Bacteria: Partial Purification and Charaterization of α-amilase by Bacillus sp. WA21. Pak. J. Bot., 43 (2) : 1045-1052.
[16] Dali, S., Arfah, A. R, Karim, A., Patong, R., 2013, Karakterisasi Enzim Amilase dari Isolat Bakteri Termofilik Bacillus subtilis, Indonesia Chemica Acta, 6 (2) : 19–26.
[17] Sivaramakrishnan S., Dhanya G., Kesavan M.N., Carlos R.S., and Ashok P. 2006. α-Amylases from Microbial Sources, Food Technol. Biotechnol. 44 (2) : 173–184.
[18] Harris, E. L. V. 1998. Concentration of Protein Extract and Purification Methods: A Practical Approach. Oxford University IRL. Press. NewYork, 123–161.
Author Information
  • Department of Chemistry, Graduate School of Science, Hasanuddin University, Makassar, South Sulawesi, Indonesia

  • Department of Chemistry, Graduate School of Science, Hasanuddin University, Makassar, South Sulawesi, Indonesia

  • Faculty of Pharmacy, Hasanuddin University, Makassar, South Sulawesi, Indonesia

  • Department of Chemistry, Graduate School of Science, Hasanuddin University, Makassar, South Sulawesi, Indonesia

  • Department of Chemistry, Graduate School of Science, Hasanuddin University, Makassar, South Sulawesi, Indonesia

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    Rugaiyah A. Arfah, Ahyar Ahmad, M. Natsir Djide, Mahdalia Anis, Muhammad Zakir. (2015). Production Optimization and Characterization of Amylase Enzyme Isolated from Termofil Bacteria Bacillus sp RSAII-1b from Lejja Hot Spring South Sulawesi. American Journal of Biomedical and Life Sciences, 3(6), 115-119. https://doi.org/10.11648/j.ajbls.20150306.13

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    ACS Style

    Rugaiyah A. Arfah; Ahyar Ahmad; M. Natsir Djide; Mahdalia Anis; Muhammad Zakir. Production Optimization and Characterization of Amylase Enzyme Isolated from Termofil Bacteria Bacillus sp RSAII-1b from Lejja Hot Spring South Sulawesi. Am. J. Biomed. Life Sci. 2015, 3(6), 115-119. doi: 10.11648/j.ajbls.20150306.13

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    AMA Style

    Rugaiyah A. Arfah, Ahyar Ahmad, M. Natsir Djide, Mahdalia Anis, Muhammad Zakir. Production Optimization and Characterization of Amylase Enzyme Isolated from Termofil Bacteria Bacillus sp RSAII-1b from Lejja Hot Spring South Sulawesi. Am J Biomed Life Sci. 2015;3(6):115-119. doi: 10.11648/j.ajbls.20150306.13

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  • @article{10.11648/j.ajbls.20150306.13,
      author = {Rugaiyah A. Arfah and Ahyar Ahmad and M. Natsir Djide and Mahdalia Anis and Muhammad Zakir},
      title = {Production Optimization and Characterization of Amylase Enzyme Isolated from Termofil Bacteria Bacillus sp RSAII-1b from Lejja Hot Spring South Sulawesi},
      journal = {American Journal of Biomedical and Life Sciences},
      volume = {3},
      number = {6},
      pages = {115-119},
      doi = {10.11648/j.ajbls.20150306.13},
      url = {https://doi.org/10.11648/j.ajbls.20150306.13},
      eprint = {https://download.sciencepg.com/pdf/10.11648.j.ajbls.20150306.13},
      abstract = {Thermostable amylase enzyme has a broad commercial value in its use in the processing of starch, sugar production, textile, paper, animal feed, pharmaceuticals and in the manufacture of detergents. This study aims to determine the optimum conditions of amylase production from the termofil bacteria Bacillus sp. RSII-1b isolated from a hot spring Lejja South Sulawesi and characterizing the amylase enzyme. The testing of amylase production was done with various concentration of starch and CaCl2 in the production medium, then fermented to obtain maximum amylase activity, amylase enzyme was produced in optimum condition, and its characteristic was tested using 2% starch substrates in various pH, temperature and determining the compound cofactor which can act as activators or inhibitors of the amylase activity, the enzyme activity was tested using DNS method. Crude extract enzyme has the highest enzyme activity of the protein content determined by the method of Lawry. The results showed that the amylase enzyme from Bacillus sp RSAII-1b isolates can be manufactured to a maximum at 33 hours of fermentation time with conditions: the concentration of substrate (starch) 1.5%, 0.08% CaCl2, 55°C temperature, medium pH 7.0 and aeration speed 200rpm with the activity of 0.1323U/mL, amylase crude extract protein content of 1.86mg/mL, with spesifik activity 0.0711 U/mg protein. Crude extract amylase work optimally at pH 6.0; 55°C -60°C the amylase activity of 0.165U/mL, the specific activity of 0.089U/mg protein. Amylase enzyme is an enzyme that depends on metal because its catalytic activity can be activated by metal ions Ca2+, Mg2+, Cu2+, Ni2+ and Co2+ as activators whereas Zn2+ ions decrease the activity of enzymes as inhibitors. Amylase activity in the crude extract optimum conditions with the addition of 10 mm ions Ca2+ can increase amylase activity up to 32,89%, while the addition of ions Zn2+ can inhibit amylase activity up to 25%.},
     year = {2015}
    }
    

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  • TY  - JOUR
    T1  - Production Optimization and Characterization of Amylase Enzyme Isolated from Termofil Bacteria Bacillus sp RSAII-1b from Lejja Hot Spring South Sulawesi
    AU  - Rugaiyah A. Arfah
    AU  - Ahyar Ahmad
    AU  - M. Natsir Djide
    AU  - Mahdalia Anis
    AU  - Muhammad Zakir
    Y1  - 2015/12/30
    PY  - 2015
    N1  - https://doi.org/10.11648/j.ajbls.20150306.13
    DO  - 10.11648/j.ajbls.20150306.13
    T2  - American Journal of Biomedical and Life Sciences
    JF  - American Journal of Biomedical and Life Sciences
    JO  - American Journal of Biomedical and Life Sciences
    SP  - 115
    EP  - 119
    PB  - Science Publishing Group
    SN  - 2330-880X
    UR  - https://doi.org/10.11648/j.ajbls.20150306.13
    AB  - Thermostable amylase enzyme has a broad commercial value in its use in the processing of starch, sugar production, textile, paper, animal feed, pharmaceuticals and in the manufacture of detergents. This study aims to determine the optimum conditions of amylase production from the termofil bacteria Bacillus sp. RSII-1b isolated from a hot spring Lejja South Sulawesi and characterizing the amylase enzyme. The testing of amylase production was done with various concentration of starch and CaCl2 in the production medium, then fermented to obtain maximum amylase activity, amylase enzyme was produced in optimum condition, and its characteristic was tested using 2% starch substrates in various pH, temperature and determining the compound cofactor which can act as activators or inhibitors of the amylase activity, the enzyme activity was tested using DNS method. Crude extract enzyme has the highest enzyme activity of the protein content determined by the method of Lawry. The results showed that the amylase enzyme from Bacillus sp RSAII-1b isolates can be manufactured to a maximum at 33 hours of fermentation time with conditions: the concentration of substrate (starch) 1.5%, 0.08% CaCl2, 55°C temperature, medium pH 7.0 and aeration speed 200rpm with the activity of 0.1323U/mL, amylase crude extract protein content of 1.86mg/mL, with spesifik activity 0.0711 U/mg protein. Crude extract amylase work optimally at pH 6.0; 55°C -60°C the amylase activity of 0.165U/mL, the specific activity of 0.089U/mg protein. Amylase enzyme is an enzyme that depends on metal because its catalytic activity can be activated by metal ions Ca2+, Mg2+, Cu2+, Ni2+ and Co2+ as activators whereas Zn2+ ions decrease the activity of enzymes as inhibitors. Amylase activity in the crude extract optimum conditions with the addition of 10 mm ions Ca2+ can increase amylase activity up to 32,89%, while the addition of ions Zn2+ can inhibit amylase activity up to 25%.
    VL  - 3
    IS  - 6
    ER  - 

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