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, Yogyakarta
2022-12-01
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| Series: | Indonesian Journal of Biotechnology |
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| Online Access: | https://jurnal.ugm.ac.id/ijbiotech/article/view/71643 |
| _version_ | 1811171035929640960 |
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| author | Dea Rizki Widiana Sotharith Phon Andriati Ningrum Lucia Dhiantika Witasari |
| author_facet | Dea Rizki Widiana Sotharith Phon Andriati Ningrum Lucia Dhiantika Witasari |
| author_sort | Dea Rizki Widiana |
| collection | DOAJ |
| 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 | 2024-04-10T17:06:48Z |
| format | Article |
| id | doaj.art-ed3f225202e043519069d77a2590d06e |
| institution | Directory Open Access Journal |
| issn | 0853-8654 2089-2241 |
| language | English |
| last_indexed | 2024-04-10T17:06:48Z |
| publishDate | 2022-12-01 |
| publisher | Universitas Gadjah Mada, Yogyakarta |
| record_format | Article |
| series | Indonesian Journal of Biotechnology |
| spelling | doaj.art-ed3f225202e043519069d77a2590d06e2023-02-06T03:00:27ZengUniversitas Gadjah Mada, YogyakartaIndonesian Journal of Biotechnology0853-86542089-22412022-12-0127421221810.22146/ijbiotech.7164333052Purification and characterization of thermostable alpha‐amylase from Geobacillus sp. DS3 from Sikidang Crater, Central Java, IndonesiaDea Rizki Widiana0Sotharith Phon1Andriati Ningrum2Lucia Dhiantika Witasari3Department of Food and Agricultural Products Technology, Faculty of Agricultural Technology, Gadjah Mada University, St. Flora No. 1, Yogyakarta, 55281, IndonesiaDepartment of Food and Agricultural Products Technology, Faculty of Agricultural Technology, Gadjah Mada University, St. Flora No. 1, Yogyakarta, 55281, IndonesiaDepartment of Food and Agricultural Products Technology, Faculty of Agricultural Technology, Gadjah Mada University, St. Flora No. 1, Yogyakarta, 55281, IndonesiaDepartment of Food and Agricultural Products Technology, Faculty of Agricultural Technology, Gadjah Mada University, St. Flora No. 1, Yogyakarta, 55281, IndonesiaAmylases 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.https://jurnal.ugm.ac.id/ijbiotech/article/view/71643alpha‐amylasethermostable enzymegeobacillus sp. ds3purificationcharacterization |
| spellingShingle | Dea Rizki Widiana Sotharith Phon Andriati Ningrum Lucia Dhiantika Witasari Purification and characterization of thermostable alpha‐amylase from Geobacillus sp. DS3 from Sikidang Crater, Central Java, Indonesia Indonesian Journal of Biotechnology alpha‐amylase thermostable enzyme geobacillus sp. ds3 purification characterization |
| 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 | alpha‐amylase thermostable enzyme geobacillus sp. ds3 purification characterization |
| url | https://jurnal.ugm.ac.id/ijbiotech/article/view/71643 |
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