Purification and characterization of thermostable serine alkaline protease from Geobacillus sp. DS3 isolated from Sikidang crater, Dieng plateau, Central Java, Indonesia
Thermostable proteases that optimally withstand the high‐temperature conditions of thermophilic bacteria could be produced and purified, which would be highly beneficial for use in industry. Geobacillus sp. is a thermophilic bacterium that can be found in various environmental conditions. The goal o...
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| Format: | Article |
| Language: | English |
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Universitas Gadjah Mada, Yogyakarta
2022-06-01
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| Series: | Indonesian Journal of Biotechnology |
| Subjects: | |
| Online Access: | https://jurnal.ugm.ac.id/ijbiotech/article/view/65822 |
| _version_ | 1828041232852451328 |
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| author | Sotharith Phon Andriati Ningrum Lucia Dhiantika Witasari |
| author_facet | Sotharith Phon Andriati Ningrum Lucia Dhiantika Witasari |
| author_sort | Sotharith Phon |
| collection | DOAJ |
| description | Thermostable proteases that optimally withstand the high‐temperature conditions of thermophilic bacteria could be produced and purified, which would be highly beneficial for use in industry. Geobacillus sp. is a thermophilic bacterium that can be found in various environmental conditions. The goal of this study was to isolate and characterize thermostable serine protease that had been produced by thermophilic Geobacillus sp. strain DS3. The proteolytic index was measured in a solid medium. The expression of protease was optimized by Geobacillus sp. DS3 at 50 °C for 18 h. Targeted protease was purified using ammonium sulfate (40‐60%) and DEAE Sephadex A‐25 resin. Using SDS‐PAGE, the molecular weight of the enzyme was predicted to be around 32 kDa. Purified thermostable protease was highly activated at 70 °C, pH 9.6 stable for 1 h, and inhibited by PMSF. Therefore, this enzyme is classified as a thermostable alkaline serine protease. Its kinetic study revealed specific activity of 0.41 U/mg (Vmax) and 0.25 mg/mL (KM). Overall, a thermostable alkaline serine protease from Geobacillus sp. DS3 showed high activity at high temperatures and alkaline pH, which is vital for application in industries such as leather processing and detergent formulation. |
| first_indexed | 2024-04-10T17:06:14Z |
| format | Article |
| id | doaj.art-696c4ebc0ed14a26b5b8559f01b51961 |
| institution | Directory Open Access Journal |
| issn | 0853-8654 2089-2241 |
| language | English |
| last_indexed | 2024-04-10T17:06:14Z |
| publishDate | 2022-06-01 |
| publisher | Universitas Gadjah Mada, Yogyakarta |
| record_format | Article |
| series | Indonesian Journal of Biotechnology |
| spelling | doaj.art-696c4ebc0ed14a26b5b8559f01b519612023-02-06T03:00:18ZengUniversitas Gadjah Mada, YogyakartaIndonesian Journal of Biotechnology0853-86542089-22412022-06-01272737910.22146/ijbiotech.6582232026Purification and characterization of thermostable serine alkaline protease from Geobacillus sp. DS3 isolated from Sikidang crater, Dieng plateau, Central Java, IndonesiaSotharith Phon0Andriati Ningrum1Lucia Dhiantika Witasari2Department of Food and Agricultural Products Technology, Faculty of Agricultural Technology, Universitas Gadjah Mada, Jl. Flora No. 1, Yogyakarta 55281, IndonesiaDepartment of Food and Agricultural Products Technology, Faculty of Agricultural Technology, Universitas Gadjah Mada, Jl. Flora No. 1, Yogyakarta 55281, IndonesiaDepartment of Food and Agricultural Products Technology, Faculty of Agricultural Technology, Universitas Gadjah Mada, Jl. Flora No. 1, Yogyakarta 55281, IndonesiaThermostable proteases that optimally withstand the high‐temperature conditions of thermophilic bacteria could be produced and purified, which would be highly beneficial for use in industry. Geobacillus sp. is a thermophilic bacterium that can be found in various environmental conditions. The goal of this study was to isolate and characterize thermostable serine protease that had been produced by thermophilic Geobacillus sp. strain DS3. The proteolytic index was measured in a solid medium. The expression of protease was optimized by Geobacillus sp. DS3 at 50 °C for 18 h. Targeted protease was purified using ammonium sulfate (40‐60%) and DEAE Sephadex A‐25 resin. Using SDS‐PAGE, the molecular weight of the enzyme was predicted to be around 32 kDa. Purified thermostable protease was highly activated at 70 °C, pH 9.6 stable for 1 h, and inhibited by PMSF. Therefore, this enzyme is classified as a thermostable alkaline serine protease. Its kinetic study revealed specific activity of 0.41 U/mg (Vmax) and 0.25 mg/mL (KM). Overall, a thermostable alkaline serine protease from Geobacillus sp. DS3 showed high activity at high temperatures and alkaline pH, which is vital for application in industries such as leather processing and detergent formulation.https://jurnal.ugm.ac.id/ijbiotech/article/view/65822thermostable serine alkaline proteasegeobacillus sp. ds3enzyme purification, deae sephadex a‐25 |
| spellingShingle | Sotharith Phon Andriati Ningrum Lucia Dhiantika Witasari Purification and characterization of thermostable serine alkaline protease from Geobacillus sp. DS3 isolated from Sikidang crater, Dieng plateau, Central Java, Indonesia Indonesian Journal of Biotechnology thermostable serine alkaline protease geobacillus sp. ds3 enzyme purification, deae sephadex a‐25 |
| title | Purification and characterization of thermostable serine alkaline protease from Geobacillus sp. DS3 isolated from Sikidang crater, Dieng plateau, Central Java, Indonesia |
| title_full | Purification and characterization of thermostable serine alkaline protease from Geobacillus sp. DS3 isolated from Sikidang crater, Dieng plateau, Central Java, Indonesia |
| title_fullStr | Purification and characterization of thermostable serine alkaline protease from Geobacillus sp. DS3 isolated from Sikidang crater, Dieng plateau, Central Java, Indonesia |
| title_full_unstemmed | Purification and characterization of thermostable serine alkaline protease from Geobacillus sp. DS3 isolated from Sikidang crater, Dieng plateau, Central Java, Indonesia |
| title_short | Purification and characterization of thermostable serine alkaline protease from Geobacillus sp. DS3 isolated from Sikidang crater, Dieng plateau, Central Java, Indonesia |
| title_sort | purification and characterization of thermostable serine alkaline protease from geobacillus sp ds3 isolated from sikidang crater dieng plateau central java indonesia |
| topic | thermostable serine alkaline protease geobacillus sp. ds3 enzyme purification, deae sephadex a‐25 |
| url | https://jurnal.ugm.ac.id/ijbiotech/article/view/65822 |
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