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

Rugaiyah A. Arfah; Ahyar Ahmad; M. Natsir Djide; Mahdalia Anis; Muhammad Zakir

doi:doi:10.11648/j.ajbls.20150306.13

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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

Rugaiyah A. Arfah, Ahyar Ahmad, M. Natsir Djide, Mahdalia Anis, Muhammad Zakir

Published in American Journal of Biomedical and Life Sciences (Volume 3, Issue 6)

Received: 22 November 2015 Accepted: 7 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%.

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

Keywords

Amylase, Hydrolytic, Bacillus sp, Isolate, Starch

References

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[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.
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[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.
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[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.
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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://article.sciencepublishinggroup.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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Author Information

Rugaiyah A. Arfah

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

Department of Chemistry, Graduate School of Science, Hasanuddin University, Makassar, South Sulawesi, Indonesia
M. Natsir Djide

Faculty of Pharmacy, Hasanuddin University, Makassar, South Sulawesi, Indonesia
Mahdalia Anis

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

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

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APA Style

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://article.sciencepublishinggroup.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}
}

Copy | Download

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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