Purification and characterization of thermostable alpha-amylase from Geobacillus sp. DS3 from Sikidang Crater, Central Java, Indonesia
Amylases are considered the most essential enzymes in biotechnology since they are widely utilized in the textile, food processing, and detergent industries. It is necessary to explore extracellular enzymatic activity in several microorganisms to discover a new potential application from amylases. I...
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| Format: | Article |
| Language: | English |
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Universitas Gadjah Mada, Research Center for Biotechnology
2022
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| Online Access: | https://repository.ugm.ac.id/282739/1/46_Purification.pdf |
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| author | Widiana, Dea Rizki Phon, Sotharith Ningrum, Andriati Witasari, Lucia Dhiantika |
| author_facet | Widiana, Dea Rizki Phon, Sotharith Ningrum, Andriati Witasari, Lucia Dhiantika |
| author_sort | Widiana, Dea Rizki |
| collection | UGM |
| description | Amylases are considered the most essential enzymes in biotechnology since they are widely utilized in the textile, food processing, and detergent industries. It is necessary to explore extracellular enzymatic activity in several microorganisms to discover a new potential application from amylases. In a previous study, thermophilic bacteria Geobacillus sp. DS3 isolated from Sikidang Crater, Dieng Plateau, Central Java, Indonesia showed amylase activity in starch medium at 70 °C. This study aimed to purify and characterize the thermostable alpha-amylase from Geobacillus sp. DS3. The alpha-amylase was produced and purified using ammonium sulfate and DEAE Sephadex A-25 column. The enzyme activity was determined using the 3,5-dinitrosalicylic acid (DNS) method. Geobacillus sp. DS3 optimally produced the alpha-amylase at 60 °C for 15 h. The alpha-amylase exhibited high enzymatic activity in 40-60 saturated ammonium sulfate extract. The molecular weight of the enzyme was estimated to be 58 kDa. The thermostable alpha-amylase showed activity at the optimum temperature of 50 °C in 200 mM sodium phosphate buffer pH 7.0. The enzyme was inhibited by EDTA, PMSF, 2-ME, and mostly by HgCl2. The Km and Vmax of the pure enzyme were 235.43 mM and 1428.57 U/mL, respectively. The result suggested that the purified thermostable alpha-amylase from Geobacillus sp. DS3 offers potential application in areas of the food industry, such as the bakery industry. |
| first_indexed | 2024-03-14T00:05:41Z |
| format | Article |
| id | oai:generic.eprints.org:282739 |
| institution | Universiti Gadjah Mada |
| language | English |
| last_indexed | 2024-03-14T00:05:41Z |
| publishDate | 2022 |
| publisher | Universitas Gadjah Mada, Research Center for Biotechnology |
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| spelling | oai:generic.eprints.org:2827392023-11-16T07:07:23Z https://repository.ugm.ac.id/282739/ Purification and characterization of thermostable alpha-amylase from Geobacillus sp. DS3 from Sikidang Crater, Central Java, Indonesia Widiana, Dea Rizki Phon, Sotharith Ningrum, Andriati Witasari, Lucia Dhiantika Food Sciences Amylases are considered the most essential enzymes in biotechnology since they are widely utilized in the textile, food processing, and detergent industries. It is necessary to explore extracellular enzymatic activity in several microorganisms to discover a new potential application from amylases. In a previous study, thermophilic bacteria Geobacillus sp. DS3 isolated from Sikidang Crater, Dieng Plateau, Central Java, Indonesia showed amylase activity in starch medium at 70 °C. This study aimed to purify and characterize the thermostable alpha-amylase from Geobacillus sp. DS3. The alpha-amylase was produced and purified using ammonium sulfate and DEAE Sephadex A-25 column. The enzyme activity was determined using the 3,5-dinitrosalicylic acid (DNS) method. Geobacillus sp. DS3 optimally produced the alpha-amylase at 60 °C for 15 h. The alpha-amylase exhibited high enzymatic activity in 40-60 saturated ammonium sulfate extract. The molecular weight of the enzyme was estimated to be 58 kDa. The thermostable alpha-amylase showed activity at the optimum temperature of 50 °C in 200 mM sodium phosphate buffer pH 7.0. The enzyme was inhibited by EDTA, PMSF, 2-ME, and mostly by HgCl2. The Km and Vmax of the pure enzyme were 235.43 mM and 1428.57 U/mL, respectively. The result suggested that the purified thermostable alpha-amylase from Geobacillus sp. DS3 offers potential application in areas of the food industry, such as the bakery industry. Universitas Gadjah Mada, Research Center for Biotechnology 2022-12 Article PeerReviewed application/pdf en https://repository.ugm.ac.id/282739/1/46_Purification.pdf Widiana, Dea Rizki and Phon, Sotharith and Ningrum, Andriati and Witasari, Lucia Dhiantika (2022) Purification and characterization of thermostable alpha-amylase from Geobacillus sp. DS3 from Sikidang Crater, Central Java, Indonesia. Indonesian Journal of Biotechnology, 27 (4). pp. 212-218. ISSN 08538654 DOI 10.22146/ijbiotech.7164 |
| spellingShingle | Food Sciences Widiana, Dea Rizki Phon, Sotharith Ningrum, Andriati Witasari, Lucia Dhiantika Purification and characterization of thermostable alpha-amylase from Geobacillus sp. DS3 from Sikidang Crater, Central Java, Indonesia |
| title | Purification and characterization of thermostable alpha-amylase from Geobacillus sp. DS3 from Sikidang Crater, Central Java, Indonesia |
| title_full | Purification and characterization of thermostable alpha-amylase from Geobacillus sp. DS3 from Sikidang Crater, Central Java, Indonesia |
| title_fullStr | Purification and characterization of thermostable alpha-amylase from Geobacillus sp. DS3 from Sikidang Crater, Central Java, Indonesia |
| title_full_unstemmed | Purification and characterization of thermostable alpha-amylase from Geobacillus sp. DS3 from Sikidang Crater, Central Java, Indonesia |
| title_short | Purification and characterization of thermostable alpha-amylase from Geobacillus sp. DS3 from Sikidang Crater, Central Java, Indonesia |
| title_sort | purification and characterization of thermostable alpha amylase from geobacillus sp ds3 from sikidang crater central java indonesia |
| topic | Food Sciences |
| url | https://repository.ugm.ac.id/282739/1/46_Purification.pdf |
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