Yudiargo, Danu Risqi (2022) Optimasi Produksi Rekombinan Bst (Bacillus Stearothermophillus) Dna Polymerase Pada Sistem Ekspresi Escherichia Coli Dalam Skala Bioreaktor. Other thesis, Institut Teknologi Sepuluh Nopember.
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Abstract
Salah satu reagen yang digunakan pada metode Loop-Mediated Isothermal Amplification (LAMP) yaitu DNA polimerase. Enzim DNA polimerase yang dapat digunakan dalam LAMP yaitu Bst DNA polimerase yang diisolasi dari bakteri termofilik Bacillus stearothermophilus. Enzim Bst DNA polimerase dapat diproduksi di dalam sel bakteri Escherichia coli dengan teknik DNA rekombinan. Escherichia coli dapat dikultivasi pada bioreaktor dengan menerapkan teknik batch fermentation. Penelitian ini bertujuan untuk melakukan optimasi produksi rekombinan Bst DNA polimerase pada sistem ekspresi Escherichia coli dalam skala bioreaktor dengan parameter yang dioptimasi yaitu agitasi dan aerasi. Dalam penelitian ini dilakukan uji ekspresi rekombinan Bst DNA polimerase dengan menggunakan Sodium Dodecyl Sulphate–Polyacrylamide Gel Electrophoresis (SDS-PAGE) yang kemudian pita protein dikuantifikasi menggunakan software ImageJ untuk mengetahui jumlah protein rekombinan. Berdasarkan uji optimasi agitasi yang telah dilakukan, diperoleh nilai agitasi optimum sebesar 100 rpm dengan konsentrasi protein dalam kultur 84,00 (ug/mL). Sedangkan pada uji optimasi aerasi, diperoleh nilai aerasi optimum sebesar 3 lpm dengan konsentrasi protein dalam kultur 84,00 (ug/mL). =========================================================================================================================
One of the reagents used in the Loop-Mediated Isothermal Amplification (LAMP) method is DNA polymerase. The DNA polymerase enzyme that can be used in LAMP is Bst DNA polymerase isolated from the thermophilic bacterium Bacillus stearothermophilus. The Bst DNA polymerase enzyme can be produced in Escherichia coli bacterial cells using recombinant DNA techniques. Escherichia coli can be cultivated in a bioreactor by applying the batch fermentation technique. This study aims to optimize the production of recombinant Bst DNA polymerase on the Escherichia coli expression system in a bioreactor scale with optimized parameters, namely agitation and aeration. In this study, the expression of recombinant Bst DNA polymerase was tested using Sodium Dodecyl Sulphate–Polyacrylamide Gel Electrophoresis (SDS-PAGE) and then the protein bands were quantified using ImageJ software to determine the amount of recombinant protein. Based on the agitation optimization test that has been carried out, the optimum agitation value of 100 rpm with protein concentration in culture was 84.00 (ug/mL). While the aeration optimization test, obtained the optimum aeration value of 3 lpm with a protein concentration in culture of 84.00 (ug/mL).
| Item Type: | Thesis (Other) |
|---|---|
| Uncontrolled Keywords: | Bst DNA Polimerase, Bioreaktor, Escherichia coli, Protein Rekombinan, Bst DNA Polymerase, Bioreactor, Escherichia coli, Recombinant Protein. |
| Subjects: | Q Science > QH Biology Q Science > QH Biology > QH601 Cell membranes Q Science > QR Microbiology |
| Divisions: | Faculty of Mathematics and Science > Biology > 46201-(S1) Undergraduate Thesis |
| Depositing User: | Eko Sulistiono |
| Date Deposited: | 30 Sep 2025 03:27 |
| Last Modified: | 30 Sep 2025 03:27 |
| URI: | http://repository.its.ac.id/id/eprint/128478 |
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