Abstract
Inulin is a naturally potential polysaccharide used to produced fructose and fructooligosaccharide. Inulinaseknown also as ß-fructosidase can hydrolise inulin to fructose or fructooligosaccharide. Inulinase production fromAspergillus niger Gmn11.1 isolated from dahlia tubers is conducted using medium containing 1% inulin and 0,2%yeast extract. The crude enzyme (filtrate culture) is purified by means of ammonium sulphate salt precipitation,followed by Sephadex G25 gel filtration column chromatography and DEAE cellulose anion exchanger columnchromatography. The result indicated that the enzyme had optimum pH and temperature of 4,6 and 450C, respectivelywith incubation time of 15 hours. The Km and Vmaxs values obtained from this experiment are 20 mg/ml and 0,769mg/ml/hours, respectively. Whereas the relative molecular weight of inulinase was monitored by SDS PAGE is 63KDa.
Highlights
naturally potential polysaccharide used to produced fructose and fructooligosaccharide
Inulinase production from Aspergillus niger Gmn11.1 isolated from dahlia tubers is conducted using medium containing
The result indicated that the enzyme had optimum pH and temperature
Summary
Aspergillus niger Gmn11.1 ditumbuhkan pada 250 ml media cair dalam erlenmeyer 500 ml. Inkubasi dilakukan pada suhu 400C dengan kecepatan agitasi 100rpm selama 5 hari. Inulinase ekstraselular dimurnikan dari filtrat kultur dengan fraksionasi amonium sulfat kemudian dilanjutkan dengan kromatografi kolom gel filtrasi sephadex G25 (3 x 70cm) Fraksi ditampung sebanyak 48 tabung masing-masing 5 ml dengan kecepatan alir 1ml/menit dan kromatografi kolom penukar ion DEAE selulosa (2 x 20cm). Aktifitas inulinase ditentukan dengan menghidrolisis larutan inulin 1% dengan inulinase (10 : 1) pada suhu 450C, pH 4,5 selama 15 jam. Gula pereduksi yang terbentuk ditentukan dengan metoda ortotoluidin, yaitu 1 ml larutan sampel ditambahkan 3 ml pereaksi ortotoluidin (94,5 ml Asam asetat glasial 5,5 ml ortotoluidin, dan 0,15 mg tiourea). Kadar protein ditentukan dengan metode Lowry et al, (1951) menggunakan BSA sebagai protein standar, dan secara langsung menggunakan spektrofotometer UV pada panjang gelombang 280 nm. Elektroforesis dijalankan dengan memakai bufer Tris glisin pH 8,3 dengan arus konstan 6 mA (Alexander et al, 1985)
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