Chrestella, Dea Jolie (2019) Visualisasi dan Aktivitas Enzim Fosfolipase A2 Bisa Calloselasma rhodostoma Boie, 1827 dan Naja sputatrix Boie, 1827 pada Variasi Suhu dan Waktu Penyimpanan. Sarjana thesis, Universitas Brawijaya.
Abstract
Enzim Fosfolipase A2 (PLA2) merupakan salah satu komponen bisa ular yang tergolong dalam kelompok toksin mayor. PLA2 memiliki profil yang cenderung stabil terhadap suhu bila dibandingkan dengan komponen protein lainnya. Penelitian ini bertujuan untuk menganalisis hasil visualisasi elektroforesis dan aktivitas enzim PLA2 bisa Calloselasma rhodostoma dan Naja sputatrix pada variasi suhu dan waktu penyimpanan. Bisa ular yang digunakan disimpan dengan perlakuan suhu sebanyak 3 tingkat yaitu -80ºC, 4ºC dan 37 ºC; dan perlakuan waktu sebanyak 3 tingkat yaitu 7, 9 dan 14 hari; serta perlakuan kontrol (tanpa perlakuan penyimpanan). Sampel bisa ular segar disentrifugasi 4000 rpm selama 5 menit (4ºC). Bagian supernatan disimpan pada kondisi yang dikehendaki. Pengukuran kadar protein dilakukan menggunakan instrumen NanoDrop. Pengujian kualitatif terhadap profil PLA2 dilakukan dengan metode Sodium Dodecyl Sulfate – Polyacrylamide Gel Elctrophoresis (SDS-PAGE). Uji aktivitas PLA2 dilakukan secara asidimetri menggunakan substrat fosfolipid dari kuning telur. Masing-masing pengujian dilakukan dengan 3 kali pengulangan. Visualisasi protein enzim PLA2 (protein pada area 13 – 16 kDa) dapat ditemui pada seluruh perlakuan penyimpanan bisa C. rhodostoma dan N. sputatrix. Data perolehan menunjukkan laju aktivitas enzim PLA2 C. rhodostoma hanya dipengaruhi oleh suhu penyimpanan (p < 0,05). Faktor suhu tidak berinteraksi dengan faktor waktu penyimpanan. Laju aktivitas enzim PLA2 N. sputatrix dipengaruhi oleh interaksi faktor suhu dan waktu penyimpanan (p < 0,05).
English Abstract
Phospholipase A2 is a component of snake venom which is classified into major toxin group. PLA2 has better thermal stability profile than the other component of snake venom because of its size and structure. The aims of this research are to analyze electrophoretic visualization and to analyze the activity of PLA2 from the venom of Calloselasma rhodostoma and Naja sputatrix in the variation of storage temperature and time. Snake venom used is stored at 10 different condition which were combination of 3 different temperature conditions: -80ºC , 4ºC and 37 ºC; and 3 different time conditions: 7, 9 and 14 days. The control group was venom solution that were examined right after secreted by milking (day 0, room temperature). Venom sample used was sentrifuged on 4000 rpm for 5 minutes at 4ºC. Supernatant was stored as desired. Protein concentration was measured by NanoDrop instrument. Qualitative assay was completed by Sodium Dodecyl Sulfate – Polyacrylamide Gel Elctrophoresis (SDS-PAGE) method to visualize the electrophoretic profile of crude venom, specifically the range of 13 – 16 kDa protein. PLA2 activity was examined by acidimetric method with the requirement of phospholipid substrate from egg yolk. Each test was done on 3 repetitions. Visualization of PLA2 (13 – 16 kDa) could be found of every venom solution that had been stored as desired. PLA2 activity of C. rhodostoma was affected by the factor of storage temperature (p < 0,05), while PLA2 activity of N. sputatrix was affected by the interaction of storage temperature and time (p < 0,05). The storage condition suggested for snake venom short storage are at cold temperature (4ºC and -80ºC) that could be last for 14 days
Other obstract
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| Item Type: | Thesis (Sarjana) |
|---|---|
| Identification Number: | SKR/MIPA/2019/197/051910921 |
| Uncontrolled Keywords: | bisa ular, fosfolipase A2, suhu penyimpanan, waktu penyimpanan, phospholipase A2, snake venom, storage temperature, storage time |
| Subjects: | 500 Natural sciences and mathematics > 536 Heat > 536.5 Temperature |
| Divisions: | Fakultas Matematika dan Ilmu Pengetahuan Alam > Biologi |
| Depositing User: | Budi Wahyono Wahyono |
| Date Deposited: | 10 Aug 2020 07:35 |
| Last Modified: | 10 Aug 2020 07:35 |
| URI: | http://repository.ub.ac.id/id/eprint/178771 |
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