PRODUKSI DAN PEMURNIAN ENZIM AMILASE DARI BAKTERI Bacillus licheniformis TS-10 MENGGUNAKAN FILTRASI GEL

PRATIWI, CITRA and Kusumawati, Heni Yohandini and Desnelli, Desnelli (2024) PRODUKSI DAN PEMURNIAN ENZIM AMILASE DARI BAKTERI Bacillus licheniformis TS-10 MENGGUNAKAN FILTRASI GEL. Undergraduate thesis, Sriwijaya University.

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Abstract

Amylase enzyme is an extracellular enzyme that can hydrolyze the internal bond of α 1,4-glucan in starch and is also one of the enzymes widely used in the industrial field. In industries that use enzymes, thermostable amylases with low substrate costs are the main choice because they can work at 60-120ºC and are usually produced by thermophilic bacteria. For certain purposes enzymes are often required in their pure state. Therefore, a pure thermostable amylase enzyme with low production cost is needed. This study aims to compare the amylase enzyme produced with soluble starch substrate and cassava peel starch from Bacillus licheniformis TS-10 bacteria and see the improvement of enzyme purity using gel filtration and protein profile analysis produced. In this study, the production of amylase enzyme from Bacillus licheniformis TS-10 bacteria using soluble starch substrate and cassava peel starch was carried out, which was then tested for enzyme activity by DNS method and protein content by Bradford method. Furthermore, the enzyme produced was purified using Sephacryl S-200 HR resin filtration gel and analyzed for protein profiles using SDS-PAGE electrophoresis and Zymography. The results showed that amylase enzyme produced using cassava peel starch substrate has specific enzyme activity that is not much different from soluble starch substrate amylase enzyme of 3.950 U/mg and has more protein bands than soluble starch substrate amylase enzyme. The amylase enzyme purified by gel filtration, which has the greatest enzyme specific activity, is found in fraction 6, which is 9.478 U/mg and has an increase in purity by 2.399 times from the crude extract of the enzyme. The protein content of fraction 6 obtained is small so that the protein band is not observed in the electrophoregram, but because the enzyme activity of the 6th fraction obtained is high in the zymogram, the clear zone is observed. The amylase enzyme purified by gel filtration which has the highest protein content is fraction 2 and 3 so that in the electrophoregram there is a protein band located around the size of BSA, namely 65 to 67 kDa.

Item Type:	Thesis (Undergraduate)
Uncontrolled Keywords:	Amylase, Starch, Gel filtration, Protein profile, Enzyme activity
Subjects:	Q Science > QD Chemistry > QD415-436 Biochemistry
Divisions:	08-Faculty of Mathematics and Natural Science > 47201-Chemistry (S1)
Depositing User:	Citra Pratiwi
Date Deposited:	23 Dec 2024 13:02
Last Modified:	23 Dec 2024 13:02
URI:	http://repository.unsri.ac.id/id/eprint/161229

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