Pengaruh Transglutaminase Terhadap Profil Protein Gelatin Kulit Ikan Patin (Pangasius hypophthalmus) Dengan Metode SDS-Page

Amalia, Yasinta Dwi (2022) Pengaruh Transglutaminase Terhadap Profil Protein Gelatin Kulit Ikan Patin (Pangasius hypophthalmus) Dengan Metode SDS-Page. Other thesis, Institut Teknologi Sepuluh Nopember.

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Abstract

Gelatin adalah protein yang diturunkan dari kolagen melalui hidrolisis parsial asam atau basa. Sumber utama gelatin berasal dari mamalia namun, gelatin dari mamalia memiliki masalah terkait larangan agama dan risiko zoonosis. Gelatin kulit ikan patin (GKIP) dapat dijadikan salah satu alternatif untuk mengatasi masalah ini namun, gelatin ikan memiliki kandungan hidroksiprolin dan prolin yang sedikit sehingga ikatan silang yang terbentuk sedikit dan berpengaruh pada sifat fisiknya. Penambahan enzim transglutaminase (TGA) dapat memperbaiki sifat fisik gelatin ikan dengan mengkatalis reaksi pembentukan ikatan silang antara gugus γ-karboksamida residu glutamin dengan gugus ε-amino residu lisin. Ikatan silang yang terbentuk menyebabkan perubahan berat molekul pada gelatin sehingga diduga dapat memengaruhi profil protein GKIP. Tujuan penelitian ini untuk mengetahui profil protein gelatin ikan patin yang diberi penambahan TGA. Kulit ikan patin diekstraksi dengan pretreatment asam dan basa. Gelatin di freeze dry lalu dibuat biofilm dan diberi perlakuan TGA 0,01 g dan TGA 0,05 g. Analisis profil protein gelatin dilakukan dengan SDS-PAGE. Hasil SDS-PAGE menemukan 11 pita protein yang diduga merupakan rantai γ (266 kDa), β (181 dan 215 kDa), subunit α1 (130 dan 140 kDa), subunit α2 (113 kDa) dan fragmen LMW (Low Molecular Weight) (96 kDa, 84 kDa, 70 kDa, 59 kDa dan 41 kDa) yang teramati pada profil protein GKIP kontrol maupun perlakuan TGA. Penambahan TGA menyebabkan terbentuknya agregat HMW (High Molecular Weight) pada bagian atas stacking gel serta intensitas komponen gelatin seperti rantai α, β dan γ yang lebih tinggi. GKIP perlakuan TGA 0,05 g memiliki intensitas pita rantai β, rantai α1 serta α2 yang lebih rendah jika dibandingkan dengan GKIP perlakuan TGA 0,01 g. Namun, intensitas rantai γ pada GKIP perlakuan TGA 0,05 g lebih tinggi dibandingkan dengan GKIP perlakuan TGA 0,01 g.
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Gelatin is a protein which is derived from collagen through partial hydrolysis with acids or bases. The main source of gelatin comes from mammals however, gelatin from mammals has problems related to religious prohibitions and zoonotic risks. Striped catfish skin gelatin (GKIP) can be used as an alternative to overcome this problem. However, fish gelatin is low in hydroxyproline and proline, so the crosslinks formed are small and affect its physical properties. The addition of the transglutaminase (TGA) enzyme can improve the physical properties of fish gelatin by catalyzing the cross-linking reaction between the γ-carboxyamide group of glutamine residue and the ε-amino group of lysine residue. The cross-links formed cause changes in the molecular weight of gelatin so that it is suspected that it can affect the protein profile of GKIP. The purpose of this study was to determine the protein profile of gelatin from striped catfish skin given the addition of TGA. The striped catfish skin was extracted by acid and base pretreatment. Gelatin was freeze dried and then made into a biofilm and treated with TGA 0.01 g and TGA 0.05 g. Gelatin protein profile analysis was performed by SDS-PAGE. The results of SDS-PAGE found 11 protein band which are suspected to be γ chains (266 kDa), β chains (181 and 215 kDa), α1 subunits (130 and 140 kDa), α2 subunits (113 kDa) and LMW (Low Molecular Weight) fragments (96 kDa, 84 kDa, 70 kDa, 59 kDa and 41 kDa) were observed in the protein profile of the control GKIP and the TGA. The addition of TGA causes the formation of HMW (High Molecular Weight) aggregates on the top of the stacking gel and the higher intensity of gelatin components such as α, β, and γ chains. GKIP TGA 0.05 g had a lower intensity of β, α1, and α2 chains when compared to GKIP TGA 0.01 g. However, the intensity of the γ chain in GKIP TGA 0.05 g was higher than in GKIP TGA 0.01 g.

Item Type:	Thesis (Other)
Additional Information:	RSBi 543 Ama p-1 2022
Uncontrolled Keywords:	Berat molekul, Gelatin, Ikan Patin, Protein, SDS-PAGE, Molecular Weight, Gelatin, Striped Catfish, Protein
Subjects:	R Medicine > RC Internal medicine > RC254 Transglutaminases. T Technology > TP Chemical technology > TP248.65.P76 Proteins--Biotechnology.
Divisions:	Faculty of Science and Data Analytics (SCIENTICS) > Biology > 46201-(S1) Undergraduate Thesis
Depositing User:	- Davi Wah
Date Deposited:	16 Jul 2024 04:48
Last Modified:	16 Jul 2024 04:48
URI:	http://repository.its.ac.id/id/eprint/108330

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