7 research outputs found
Identifikasi Gelatin dalam Beberapa Obat Bentuk Sediaan Tablet Menggunakan Metode Spektrofotometri
Get PDFPurification and Characterization of Fibrinolytic Proteases from Mushroom Volvariela volvaceae Extract.
No full textPurification and Characterization of Fibrinolytic Proteases from Mushroom Volvariela volvaceae Extract. Ediblestraw mushroom (V. volvaceae) has been known used for improvement of blood circulation due to its fibrinolyticcontent. The objective of the study is to purify and characterize fibrinolytic protease from straw mushroom extract.Purification were performed through several steps, i.e. precipitation using ammonium sulphate 75%, dialyzed membran(cut-off 10 kDa), and ion-exchange chromatography using DEAE Sepharose. The active fraction of DEAE-Sepharosecontains two purified protein bands with molecular weight of 12.9 and 15.8 kDa. The active fraction has specificactivity of 0.383 U/mg with 2.7 fold higher purification compared to its crude extract. Both crude and purified enzymeshad optimum activity at temperature of 50 ºC and pH 7 in 10 minutes of incubation. Fibrin zymographic profiledemonstrated that the enzyme hydrolyzed fibrin, as well as casein, indicating their potent fibrinolytic activity. Theenzyme was strongly inhibited by phenilmethylsulphonyl fluoride and N-p-tosil-L-lysinchloromethyl keton. Thissuggested that it was a serine protease. In summary, these results showed that crude and purified protease of strawmushroom (V. volvaceae) has fibrinolytic activities that can be applied for alternative thrombolytic therapy
Purifikasi Dan Pencirian Enzim Protease Fibrinolitik Dari Ekstrak Jamur Merang
Get PDFPurification and Characterization of Fibrinolytic Proteases from Mushroom Volvariela volvaceae Extract. Edible straw mushroom (V. volvaceae) has been known used for improvement of blood circulation due to its fibrinolytic content. The objective of the study is to purify and characterize fibrinolytic protease from straw mushroom extract. Purification were performed through several steps, i.e. precipitation using ammonium sulphate 75%, dialyzed membran (cut-off 10 kDa), and ion-exchange chromatography using DEAE Sepharose. The active fraction of DEAE-Sepharose contains two purified protein bands with molecular weight of 12.9 and 15.8 kDa. The active fraction has specific activity of 0.383 U/mg with 2.7 fold higher purification compared to its crude extract. Both crude and purified enzymes had optimum activity at temperature of 50 ºC and pH 7 in 10 minutes of incubation. Fibrin zymographic profile demonstrated that the enzyme hydrolyzed fibrin, as well as casein, indicating their potent fibrinolytic activity. The enzyme was strongly inhibited by phenilmethylsulphonyl fluoride and N-p-tosil-L-lysinchloromethyl keton. This suggested that it was a serine protease. In summary, these results showed that crude and purified protease of straw mushroom (V. volvaceae) has fibrinolytic activities that can be applied for alternative thrombolytic therapy
Silkworm (Bombyx mori) hemolymph unable to substitute fetal bovine serum in insect cell culture
No full textPURIFIKASI DAN PENCIRIAN ENZIM PROTEASE FIBRINOLITIK DARI EKSTRAK JAMUR MERANG
Get PDFPurification and Characterization of Fibrinolytic Proteases from Mushroom Volvariela volvaceae Extract. Edible straw mushroom (V. volvaceae) has been known used for improvement of blood circulation due to its fibrinolytic content. The objective of the study is to purify and characterize fibrinolytic protease from straw mushroom extract. Purification were performed through several steps, i.e. precipitation using ammonium sulphate 75%, dialyzed membran (cut-off 10 kDa), and ion-exchange chromatography using DEAE Sepharose. The active fraction of DEAE-Sepharose contains two purified protein bands with molecular weight of 12.9 and 15.8 kDa. The active fraction has specific activity of 0.383 U/mg with 2.7 fold higher purification compared to its crude extract. Both crude and purified enzymes had optimum activity at temperature of 50 ºC and pH 7 in 10 minutes of incubation. Fibrin zymographic profile demonstrated that the enzyme hydrolyzed fibrin, as well as casein, indicating their potent fibrinolytic activity. The enzyme was strongly inhibited by phenilmethylsulphonyl fluoride and N-p-tosil-L-lysinchloromethyl keton. This suggested that it was a serine protease. In summary, these results showed that crude and purified protease of straw mushroom (V. volvaceae) has fibrinolytic activities that can be applied for alternative thrombolytic therap
PURIFIKASI DAN PENCIRIAN ENZIM PROTEASE FIBRINOLITIK DARI EKSTRAK JAMUR MERANG
No full textJamur merang (Volvariela volvaceae) merupakan jamur pangan yang secara empiris diduga dapat melancarkan peredaran darah sehubungan dengan kandungan enzim fibrinolitik yang dimilikinya. Penelitian ini bertujuan untuk mendapatkan enzim protease murni yang bersifat fibrinolitik dan penciriannya dari ekstrak jamur merang. Pemurnian dilakukan secara bertahap, yaitu presipitasi amonium sulfat 75%, dialisis dengan menggunakan membran dialisis (cutoff 10 kDa), dan kromatografi pertukaran ion dengan matriks DEAE Sepharose. Fraksi eluat DEAE-Sepharose aktif mempunyai dua subunit protein dengan bobot molekul 12,9 dan 15,8 kDa. Eluat memiliki aktivitas spesifik 0,383 U/mg dengan tingkat kemurniannya mencapai 2,7 kali lebih tinggi dibandingkan dengan ekstrak kasarnya. Analisis pencirian enzim dilakukan terhadap ekstrak kasar dan eluat murni. Aktivitas ekstrak kasar dan eluat mencapai optimum pada suhu 50 ºC dan pH 7 dengan waktu inkubasi 10 menit. Pada kondisi optimum, aktivitas protease eluat lebih besar daripada ekstrak kasarnya. Enzim dihambat kuat oleh inhibitor fenilmetilsulfonil fluorida dan N-p-tosil-L-lisinklorometil keton yang menunjukkan golongan protease serin. Profil zimografi fibrin memperlihatkan bahwa enzim mampu mendegradasi substrat fibrin. Hasil ini membuktikan bahwa ekstrak dan eluat protease murni dari jamur merang (V. volvaceae) mempunyai aktivitas fibrinolitik yang potensial dan dapat dimanfaatkan sebagai alternatif terapi trombolitik
