Home » Volume 1 Nomor 1 » PROTEIN BIOAKTIF DARI BAGIAN TANAMAN Coccinia grandis (L.) Voigh DAN AKTIVITASNYA TERHADAP GALUR SEL KANKER

PROTEIN BIOAKTIF DARI BAGIAN TANAMAN Coccinia grandis (L.) Voigh DAN AKTIVITASNYA TERHADAP GALUR SEL KANKER

Author: Churiyah dan Said Harran

Pusat Teknologi Farmasi dan Medika, Badan Pengkajian dan Penerapan Teknologi
Departemen Biologi,  FMIPA, Institut Pertanian Bogor
 

ABSTRACT

The research was conducted to isolate and activity analysis of bioactive protein from (Coccinia grandis (L.) Voigh,. Plants was extracted with phosphate buffer saline (PBS), than the protein was precipitated using 80% saturated ammonium sulphate, continued with the dialysis  using pH 7 phosphate buffer. The dialysate was fractionated thorough gel filtration chromatography and characterized using SDS-PAGE. The toxicity of protein was analyzed through brine shrimp lethality test (BSLT) and followed with cytotoxicity test using HeLa and K-562 cancer cell line. Three bioactive protein were isolated from the mature fruits, the  immature fruit and the root. The protein yield were respectively 0.023 % from the immature fruit, 0.012 % from mature fruit and 0.109 % from  the root. All of the protein were toxic on BSLT with LC50  within the range of  23 -32 µg/ml. Characterization of  protein using SDS-PAGE indicated the molecular masses of those protein were approximately  19-41kDa. The cytotoxicity test of the immature fruit protein fraction using cancer cell line showed that the addition of  this protein could inhibit proliferation of HeLa cell up to 34.4% and K-562 cell  up to 40.98% compared to that of non treated cell..

Keywords : Coccinia grandis (L.) Voigh, protein, toxicity

 

ABSTRAK

Penelitian ini bertujuan mengisolasi dan menganalisis aktivitas protein bioaktif dari (Coccinia grandis (L.) Voigh. Tanaman diekstraksi dengan buffer fosfat saline (PBS), proteinnya diendapkan dengan amonium sulfat kejenuhan 80%, dilanjutkan dengan dialisis menggunakan  buffer fosfat pada pH 7. Dialisat difraksinasi dengan kromatografi filtrasi gel dan dikarakterisasi menggunakan SDS-PAGE. Toksisitas protein dianalisis melalui uji brine shrimp lethality test (BSLT) dan dilanjutkan dengan uji sitotoksisitas menggunakan HeLa (human service carcinoma) dan sel K-562 (human erythro leukemia). Tiga protein bioaktif diisolasi dari buah matang, buah belum matang dan akar. Hasil protein yang dilakukan masing-masing 0,023% dari buah belum matang, 0,012% dari buah matang dan 0,109% dari akar. Semua protein yang dihasilkan beracun pada BSLT dengan LC50 dalam kisaran 23-32 ug/ml. Karakterisasi protein dengan SDS-PAGE menunjukkan massa molekul protein tersebut adalah sekitar 19-41 kDa. Uji sitotoksisitas dari fraksi protein buah yang belum matang menggunakan galur sel kanker menunjukkan bahwa  protein ini dapat menghambat proliferasi sel HeLa hingga 34,4% dan sel K-562 hingga 40,98%.

Kata kunci : Coccinia grandis (L.) Voigh, protein, toksisitas.

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