SKRINING RIBOSOME INACTIVATING PROTEIN (RIP) DARI BERBAGAI SAYUR SERTA KAJIAN STABILITAS PROTEIN DAN AKTIVITASNYA SELAMA PEREBUSAN

GABRIELA KASIH M, Dr. Rumiyati, S.Si., M.Si., Apt.; Prof. Dr. Sismindari, SU., Apt.

2014 | Skripsi | FARMASI

Ribosome Inactivating Protein (RIP) adalah protein yang mempunyai aktivitas enzimatik N-glikosidase sehingga mampu menghambat sintesis protein. RIP mempunyai sifat sitotoksik terhadap sel mamalia serta aktivitas lain seperti antivirus, antibakteri, antioksidan, fotoprotektif, dan kemopreventif. RIP ditemukan pada tanaman termasuk tanaman yang dapat dimakan seperti sayur yang merupakan sumber pangan sehari-hari. Oleh karena itu, perlu dilakukan identifikasi RIP pada sayuran yang sering dikonsumsi. Skrining keberadaan RIP pada ekstrak gubal protein sayur dilakukan dengan uji aktivitas pemotongan DNA superkoil untuk mengetahui tanaman yang menjadi bahan pangan di Indonesia yang mengandung RIP. Stabilitas dan aktivitas RIP dalam sayur diuji dengan perebusan selama 5 menit. Uji sitotoksisitas pada sel kanker payudara MCF-7 dengan metode MTT dilakukan untuk mengetahui aktivitas sitotoksiknya terhadap sel mamalia. Hasil skrining keberadaan RIP pada sayur buncis (Phaseolus vulgaris L.), kacang panjang (Vigna sinensis L.), kenikir (Cosmos caudatus Kunth.), bayam (Amaranthus hybridus L.) dan leunca (Solanum nigrum L.) menunjukkan bahwa kelima sayur tersebut mengandung RIP. RIP di dalam buncis cukup stabil oleh perlakuan perebusan selama 5 menit, sedangkan stabilitas RIP di dalam kacang panjang rebus berkurang. Ekstrak gubal kenikir mempunyai aktivitas sitotoksik terhadap sel MCF-7 dengan IC50 sebesar 4,44 mg/mL.RIP pada ekstrak gubal buncis, dan kacang panjang memiliki aktivitas sitotoksik yang rendah terhadap kultur sel MCF7 dan data IC50 belum dapat dihitung karena prosentase kematian tertingginya berturut-turut hanya mencapai 14,62% dan 17,78%.

Ribosome Inactivating Protein (RIP) is a kind of protein which has enzymatic activity named N-glycosidase that can inhibit the protein synthesis. RIP has cytotoxic characteristic against mammalian cells as well as other activities such as antiviral, antibacterial, antioxidant, photoprotective, and chemopreventive. RIP can be found in various plants, including edible plants like vegetables, which is a source of daily food. Therefore, it is necessary to identify RIP in ordinary consumed vegetables. Screening the presence of RIP in the crude extract protein of vegetable is performed by testing the DNA supercoiled-cleaving activity in order to determine which plants that are used as Indonesian daily food may contains RIP. Stability and activity of RIP in vegetables is tested by stewing it for 5 minutes. Cytotoxicity assay toward MCF-7 breast cancer cells using MTT assay is performed to determine its cytotoxic activity against mammalian cells. The results of screening the presence of RIP in green bean (Phaseolus vulgaris L.), long bean (Vigna sinensis L.), Kenikir leaves (Cosmos caudatus Kunth.), Chinese amaranth leaves (Amaranthus hybridus L.) and Leunca (Solanum nigrum L.) showed that all these vegetables have RIP. RIP in green beans is fairly stable after stew it for 5 minutes, while the stability of the RIP is reduced in stewed long beans. Crude extract of kenikir leaves have cytotoxic activity against MCF-7 cells with IC50 of 4.44 mg/mL. RIP in crude extracts of long beans and green beans have low cytotoxic activity against MCF-7 cell culture and IC50 value cannot be calculated because the highest death percentage only reached respectively in 14.62% and 17.78%.

Kata Kunci : Ribosome Inactivating Protein, sayuran, perebusan, DNA superkoil, sitotoksisitas, sel MCF-7