PRODUKSI ENZIM PROTEASE DARI ISOLAT BAKTERI Bacillus licheniformis TS-17 DAN PEMURNIAN MENGGUNAKAN FRAKSINASI AMONIUM SULFAT DAN KROMATOGRAFI GEL FILTRASI

ANNISA, SITI FATH and Kusumawati, Heni Yohandini (2024) PRODUKSI ENZIM PROTEASE DARI ISOLAT BAKTERI Bacillus licheniformis TS-17 DAN PEMURNIAN MENGGUNAKAN FRAKSINASI AMONIUM SULFAT DAN KROMATOGRAFI GEL FILTRASI. Undergraduate thesis, Sriwijaya University.

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Abstract

Protease is an enzyme that can catalyze hydrolysis reactions to break down protein molecules into peptides and amino acids. Microorganisms, such as fungi, yeast, and bacteria can produce proteases. One of the bacteria that can produce protease is Bacillus licheniformis TS-17 isolated from the Tanjung Sakti Lahat Hot Springs. The protease produced was a thermostable protease resistant to high temperatures and pH so it is useful for the leather industry. This study carried out purifying enzymes using ammonium sulfate fractionation and gel filtration chromatography, expressing the extracellular protein profile using SDS-PAGE, and determining the enzyme activity towards substrates using zymography. B. licheniformis TS-17 was grown on chicken feather waste and skim milk. After the enzymes from both media were produced, activity tests with the Folin-Ciocalteu reagent and determination of protein concentration with the Bradford reagent were carried out to compare the specific activities of the enzymes from both media. Chicken feather waste media enzymes with higher specific activity go through the first purification stage in the form of ammonium sulfate fractionation. After the fraction with the highest specific activity was identified as a result of ammonium sulfate fractionation, the fraction was subjected to a second stage of purification in the form of gel filtration chromatography. Crude extracts of chicken feather waste media enzymes, crude extracts of skim milk enzymes, enzymes resulting from ammonium sulfate fractionation, and enzymes resulting from gel filtration chromatography purification were characterized for their protein profile using SDS-PAGE and their activity towards casein using Zymography. This study showed that the crude extract of chicken feather waste media enzymes had a higher specific activity of 2.5541 U/mg compared to the crude extract of skim milk media enzymes of 2.1326 U/mg. Fraction 3 from ammonium sulfate had the highest specific activity of 6.0264 U/mg. Fraction 6 from gel filtration chromatography had the highest specific activity of 6.7137 U/mg. The purity of fraction 3 by ammonium sulfate fractionation increased by 1.88 times from its crude extract and fraction 6 by gel filtration chromatography increased by 2.09 times from its crude extract. Extracellular protein profile expressed by B. licheniformis TS-17 in chicken feather waste media is not the same as in skim milk media. The proteins expressed by B. licheniformis TS-17 in the crude enzyme extract, the enzyme after ammonium sulfate fractionation, and the enzyme after gel filtration chromatography have protease activity against casein, but the proteins’ sizes differ.

Item Type:	Thesis (Undergraduate)
Uncontrolled Keywords:	Bacillus licheniformis TS-17, Chicken Feather Medium, Skim Milk Medium, Gel Filtration Chromatography, SDS-PAGE, Zymography
Subjects:	Q Science > QD Chemistry > QD415-436 Biochemistry
Divisions:	08-Faculty of Mathematics and Natural Science > 47201-Chemistry (S1)
Depositing User:	Siti Fath Annisa
Date Deposited:	23 Dec 2024 13:03
Last Modified:	23 Dec 2024 13:03
URI:	http://repository.unsri.ac.id/id/eprint/161253

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