Muhadzib, Hasna and Douglas J H Shyu, PhD and Aji Sutrisno, PhD (2023) Identification of Curdlan (Beta-1,3-Glucan) Binding Proteins from Lyophilized Gac Fruit Aril (Momordica cochinchinensis). Sarjana thesis, Universitas Brawijaya.
Abstract
Studi ini bertujuan untuk mengidentifikasi protein pengikat curdlan (beta-1,3- glukan) dari aril buah gac (Momordica cochinchinensis) yang diekstraksi melalui metode pengeringan beku. Pengeringan beku dievaluasi menjadi metode ekstraksi yang paling efektif dari metode ekstraksi aril buah lainnya berdasarkan konsentrasi protein dari sampel yang dihasilkan. Metode purifikasi untuk aril buah gac juga dikembangkan dengan metode purifikasi berdasarkan afinitas menggunakan curdlan beads. Konsentrasi protein dari ekstrak aril buah gac diidentifikasi cukup tinggi melalui metode ELISA yaitu sebesar 5277 mg/ml . Analisis SDS-PAGE memperlihatkan adanya aktivitas protein pada berat molekul 45, 32, 23, 21, dan 19 kDa. Protein tersebut adalah protein yang berhasil terikat dengan curdlan. Melalui uji zymography, terlihat pula aktivitas proteolitik pada berat molekul ~45 kDa. Berdasarkan hasil tersebut, telah diidentifikasi protein dengan berat molekul 45, 32, dan 21 kDa. Protein tersebut dipilih berdasarkan potensi manfaat dari protein dengan berat molekul tersebut. Analisis LC-MS mengidentifikasi pita 45 kDa (skor: 49) sebagai protein desiccation-related protein PCC13-62-like, yang dikenal karena perannya dalam mencegah dehidrasi saat kehilangan air yang ekstrem pada tumbuhan. Sedangkan pita 32 kDa (skor: 72) diidentifikasi sebagai chitinase, yang berfungsi sebagai agen antifungi pada tumbuhan. Identifikasi lebih lanjut dari pita 21 kDa sedang dilakukan melalui metode N-terminal Sequence Analysis untuk meningkatkan akurasi identifikasi.
English Abstract
This study aimed to identify curdlan (beta-1,3-glucan) binding proteins from lyophilized gac fruit aril (Momordica cochinchinensis). Different methods of fruit aril extraction, including freeze-drying, air drying, and spray-drying, were evaluated, with freeze-drying yielding the highest protein activity. The purification method of the fruit aril also developed using affinity purification with curdlan beads. The purified protein solution concentration was determined to be 5.277 mg/ml which considered high. SDS-PAGE analysis revealed protein bands of 45, 32, 23, 21, and 19 kDa that successfully bound to curdlan, while zymography indicated proteolytic activity around ~45 kDa. Due to budget constraints, only three bands were identified based on their potential benefits and molecular weights. LC-MS analysis identified the 45 kDa (score : 49) band as a desiccation-related protein PCC13-62-like, known for its role in desiccation tolerance, preventing dehydration during water loss in plants. The 32 kDa (score : 72) band was identified as chitinase, which serves as an antifungal agent in plants. Further identification of the 21 kDa band is ongoing, utilizing N-terminal sequence analysis and additional amino acid sequence databases for improved identification.
| Item Type: | Thesis (Sarjana) |
|---|---|
| Identification Number: | 052310 |
| Uncontrolled Keywords: | Chitinase, Curdlan Binding Proteins, Desiccation-Related Protein, Freeze-Drying, Gac Fruit (Momordica cochinchinensis) Aril, Aril Buah Gac (Momordica cochinchinensis), Chitinase, DesiccationRelated Protein, Pengeringan Beku, Protein Pengikat Curdlan, |
| Divisions: | Fakultas Teknologi Pertanian > Teknologi Hasil Pertanian |
| Depositing User: | Annisti Nurul F |
| Date Deposited: | 16 Jan 2024 02:02 |
| Last Modified: | 16 Jan 2024 02:02 |
| URI: | http://repository.ub.ac.id/id/eprint/210575 |
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