Riani, Catur
Sekolah Farmasi, Institut Teknologi Bandung, Jalan Ganesha 10 Bandung 40132

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Optimasi Isolasi dan Karakterisasi Jakalin dari Biji Nangka Suciati, Tri; Widanengsih, Niknik; Riani, Catur; Gusdinar, Tutus
Acta Pharmaceutica Indonesia Vol 37, No 4 (2012)
Publisher : School of Pharmacy Institut Teknologi Bandung

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (606.272 KB)

Abstract

Telah diisolasi jakalin dari biji nangka (Artocarpus heterophyllus) menggunakan metode kromatografi afinitas dengan matriks guar gum yang telah dipaut silang dengan epiklorohidrin, isolasi dilakukan dengan menggunakan pengelusi D-galaktosa. Hasil karakterisasi jakalin menggunakan SDS-PAGE menunjukkan bahwa jakalin memiliki dua pita dengan bobot molekul 14,1 dan 15,5 kDa, antar subunitnya tidak dihubungkan dengan ikatan disulfida. Jakalin yang diisolasi memiliki kemampuan mengaglutinasi eritrosit, kemampuan hemaglutinasi jakalin tidak berkurang setelah diinkubasi pada suhu 20oC dan 30oC, tetapi setelah inkubasi pada suhu 40oC, 50oC, 60oC, 70oC, aktivitasnya turun masing-masing 75%, 87,5%, 93,75% dan 98,4%. Kemampuan hemaglutinasi jakalin tidak berkurang setelah diinkubasi pada pH 5, 6, dan 7, tetapi aktivitasnya turun 75% setelah inkubasi pada pH 2, 3, 4, dan 8, serta turun 96,9% pada pH 9 dan 10. Kemampuan hemaglutinasi jakalin dihambat oleh D-galaktosa dengan kemampuan inhibisi hemaglutinasi sebesar 6,25 mM, aktivitas hemaglutinasi jakalin tidak dihambat oleh D-manosa, D-glukosa, fruktosa, laktosa, arabinosa, maltosa, dan manitol. Diperoleh jakalin dengan perolehan rata-rata sebesar 0,32% b/b dari serbuk kering biji nangka.Kata kunci: Jakalin, lektin, Artocarpus heterophyllus, D-galaktosa.Jacalin from jackfruit (Artocarpus heterophyllus) had been isolated using affinity chromatography method with epichlorohydrin crosslinked guar gum as the matrix, isolation carried out using D-galactose as the eluen. Characterizationjacalin using SDS-PAGE showed that jacalin has two bands with molecular weights 14.1 and 15.5 kDa, intersubunit not connected with disulfide bonds. Jacalin isolates have the ability to haemagglutination erythrocytes, haemagglutination activity of jacalin maintained after incubation at 20oC and 30oC, but the activity decreased at 40, 50, 60, and 70 oC of incubation, which were 75%, 87.5%, 93.75% and 98.4% , respectively. Jacalin hemagglutination ability is not reduced after incubation at pH 5, 6, and 7, but the activity down 75% after incubation at pH 2, 3, 4, and 8, and down 96.9% at pH 9 and 10. The haemagglutination activity of jacalin was inhibited by D-galactose with the capacity inhibition value was 6.25 mM, but it was not inhibited by D-manose, D-glucose, fructose, lactose, arabinose, maltose, and manitol. Average recovery of jacalin is 0.32% w/w of jackfruit dry powder.Keywords: Jacalin, lectin, Artocarpus heterophyllus, D-galactose.
Optimasi Isolasi dan Karakterisasi Jakalin dari Biji Nangka Tri Suciati; Niknik Widanengsih; Catur Riani; Tutus Gusdinar
Acta Pharmaceutica Indonesia Vol. 37 No. 4 (2012)
Publisher : School of Pharmacy Institut Teknologi Bandung

Show Abstract | Download Original | Original Source | Check in Google Scholar

Abstract

Telah diisolasi jakalin dari biji nangka (Artocarpus heterophyllus) menggunakan metode kromatografi afinitas dengan matriks guar gum yang telah dipaut silang dengan epiklorohidrin, isolasi dilakukan dengan menggunakan pengelusi D-galaktosa. Hasil karakterisasi jakalin menggunakan SDS-PAGE menunjukkan bahwa jakalin memiliki dua pita dengan bobot molekul 14,1 dan 15,5 kDa, antar subunitnya tidak dihubungkan dengan ikatan disulfida. Jakalin yang diisolasi memiliki kemampuan mengaglutinasi eritrosit, kemampuan hemaglutinasi jakalin tidak berkurang setelah diinkubasi pada suhu 20oC dan 30oC, tetapi setelah inkubasi pada suhu 40oC, 50oC, 60oC, 70oC, aktivitasnya turun masing-masing 75%, 87,5%, 93,75% dan 98,4%. Kemampuan hemaglutinasi jakalin tidak berkurang setelah diinkubasi pada pH 5, 6, dan 7, tetapi aktivitasnya turun 75% setelah inkubasi pada pH 2, 3, 4, dan 8, serta turun 96,9% pada pH 9 dan 10. Kemampuan hemaglutinasi jakalin dihambat oleh D-galaktosa dengan kemampuan inhibisi hemaglutinasi sebesar 6,25 mM, aktivitas hemaglutinasi jakalin tidak dihambat oleh D-manosa, D-glukosa, fruktosa, laktosa, arabinosa, maltosa, dan manitol. Diperoleh jakalin dengan perolehan rata-rata sebesar 0,32% b/b dari serbuk kering biji nangka.Kata kunci: Jakalin, lektin, Artocarpus heterophyllus, D-galaktosa.Jacalin from jackfruit (Artocarpus heterophyllus) had been isolated using affinity chromatography method with epichlorohydrin crosslinked guar gum as the matrix, isolation carried out using D-galactose as the eluen. Characterizationjacalin using SDS-PAGE showed that jacalin has two bands with molecular weights 14.1 and 15.5 kDa, intersubunit not connected with disulfide bonds. Jacalin isolates have the ability to haemagglutination erythrocytes, haemagglutination activity of jacalin maintained after incubation at 20oC and 30oC, but the activity decreased at 40, 50, 60, and 70 oC of incubation, which were 75%, 87.5%, 93.75% and 98.4% , respectively. Jacalin hemagglutination ability is not reduced after incubation at pH 5, 6, and 7, but the activity down 75% after incubation at pH 2, 3, 4, and 8, and down 96.9% at pH 9 and 10. The haemagglutination activity of jacalin was inhibited by D-galactose with the capacity inhibition value was 6.25 mM, but it was not inhibited by D-manose, D-glucose, fructose, lactose, arabinose, maltose, and manitol. Average recovery of jacalin is 0.32% w/w of jackfruit dry powder.Keywords: Jacalin, lectin, Artocarpus heterophyllus, D-galactose.
Enzymatic Characterization of Recombinant Cyclodextrin Glycosyltransferase from Bacillus sp. a2-5a using Sagoo Starch as Substrate RINA IMANIAR; CATUR RIANI; DESSY NATALIA; DEBBIE SOFFIE RETNONINGRUM
Microbiology Indonesia Vol. 6 No. 3 (2012): September 2012
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (485.698 KB) | DOI: 10.5454/mi.6.3.5

Abstract

Cyclodextrin (CD) is a cyclic oligosaccharide molecule and depending on the number of glucose molecules, three types of CDs are commonly used, α-CD, β-CD, and γ-CD. CDs can be produced enzymatically using starch as substrate catalyzed by CD glycosyltransferase (CGTase). In current research, recombinant CGTase production from the synthetic gene was optimized for its production using three growth media and two induction temperatures. The highest yield was obtained in Luria Bertani medium at 25 °C. The rCGTase protein was affinity purified as a 76.39 kDa protein which showed α-cyclization and starch hydrolysis activities using zymography method. The optimum temperature, pH and incubation time was 55 °C, 6, and 24 h, respectively. The enzyme was stable at a wide pHs in the range of 5-10, retained its half activity at 56 °C for 30 min and had cyclization ratio for α-CD : β-CD : γ-CD was 4 : 81 : 15. An amount of 542 mg β-cyclodextrin was produced from 100 mL reaction of 1% (b/v) sagoo starch using 38.4 μg rCGTase in optimum condition. This work reports for the first time the character of rCGTase from Bacillus sp. A2-5a using sagoo starch as a substrate.