Deteksi Aktivitas Fibrinolitik Isolat Bakteri WU 021055* Asal Perairan Pantai Papuma Jember Menggunakan Zimografi

Abstract

Bakteri merupakan sumber penting berbagai enzim termasuk enzim fibrinolitik. Enzim ini diperlukan untuk mendegradasi bekuan darah pada orang yang mengalami penyakit trombosis. Isolat bakteri WU 021055* asal Pantai Papuma Jember terbukti menghasilkan enzim fibrinolitik ekstraseluler. Penelitian ini bertujuan mengetahui ukuran protein yang memiliki aktivitas fibrinolitik dan mengidentifikasi karakteristik morfologi isolat WU bakteri WU 021055*. Penelitian ini dilakukan di Laboratorium Mikrobiologi, Fakultas Matematika dan Ilmu Pengetahuan Alam, Universitas Jember pada April–Agustus 2014. Aktivitas fibrinolitik presipitat protein (PP) ditentukan menggunakan metode fibrin plate agar dan zimografi fibrin. Ekstrak protein kasar (EPK) dipanen pada jam ke-12 dan dipresipitasi menggunakan amonium sulfat 80%. Hasil uji aktivitas fibrinolitik menggunakan fibrin plate agar menunjukkan presipitat memiliki aktivitas fibrinolitik lebih besar dibanding dengan EPK. Dari hasil karakterisasi PP menggunakan sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) diperoleh 11 pita protein dengan ukuran 12–41 kDa. Berdasar atas hasil zimografi fibrin, pita protein dengan berat molekul 24 kDa yang memberikan aktivitas fibrinolitik. Protein dengan ukuran 24 kDa ini mampu mendegradasi substrat fibrin. Simpulan, isolat bakteri WU 021055* mengandung berbagai protein ekstraseluler, memiliki bentuk koloni bulat berwarna putih dan termasuk bakteri gram prositif berbentuk batang.

DETECTION OF FIBRINOLYTIC ACTIVITY OF WU 021055* BACTERIAL ISOLATE FROM PAPUMA BEACH COASTAL JEMBER USING ZYMOGRAPHY

Bacteria were important resources for various enzymes including fibrinolytic enzymes. This enzyme is  capable of degrading fibrin clot in patient with thrombotic diseases. Bacterial isolate of WU 021055* from Papuma Beach Coastal Jember could secrete extracellular fibrinolytic enzymes. The objective of this reasearch was to determine the molecular weight of protein responsible for fibrinolytic activity and to identify morphologycal characterization of bacterial isolate of WU 021055*. This study was conducted at Laboratory of Microbiology, Faculty of Mathematics and Natural Sciences, Universitas Jember in April–August 2014. Fibrinolytic activity of precipitate protein (PP) was determined by using fibrin plate agar and fibrin zymography. Crude protein extract (CPE) was harvested at 12 hours and precipitated by 80% ammonium sulphates. The result of fibrinolityc activity determination showed that fibrinolytic activity of PP was higher than CPE. Protein characterization of PP by using sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) obtained 11 different protein bands corresponds to value 12–42 kDa. Based on fibrin zymography, the 24 kDa protein might contribute to fibrinolytic activity due to degraded fibrin substrates. In conclusion, bacterial isolate of WU 021055* contained extracellular fibrin protein was white colony and gram positives bacilli able to degraded.