[논문]Characterization of Two Forms of Glucoamylase from Traditional Korean Nuruk Fungi, Aspergillus coreanus NR 15-1

HAN YOUNG JIN; YU TAE SHICK

Characterization of Two Forms of Glucoamylase from Traditional Korean Nuruk Fungi, Aspergillus coreanus NR 15-1 원문보기

Journal of microbiology and biotechnology, v.15 no.2, 2005년, pp.239 - 246

HAN YOUNG JIN (Department of Microbiology, College of Natural Science, Keimyung University) , YU TAE SHICK (Department of Microbiology, College of Natural Science, Keimyung University)

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Some characteristics of two forms of glucoamylase (glucan 1 A-$\alpha$-glucosidase, EC 3. 2. I. 3) purified from Aspergillus coreanus NR 15-1 were investigated. The enzymes were produced on a solid, uncooked wheat bran medium of A. coreanus NR 15-1 isolated from traditional Korean Nuruk. Two forms of glucoamylase, GA-I and GA-II, were purified to homogenity after 5.8-fold and 9.6-fold purification, respectively, judged by disc- and SDS-polyacrylamide gel electrophoresis. The molecular mass of GA-I and GA-II were estimated to be 62 kDa and 90 kDa by Sephadex G-1OO gel filtration, and 64 kDa and 91 kDa by SDS-polyacrylarnide gel electrophoresis, respectively. The optimum temperatures of GA-I and GA-II were 60$^circ$C and 65$^circ$C, respectively, and the optimum pH was 4.0. The activation energy (Ea value) of GA-I and GA-II was 11.66 kcal/mol and 12.09 kcal/mol, respectively, and the apparent Michaelis constants (K_{m}) of GA-I and GA-II for soluble starch were found to be 3.57 mg/ml and 6.25 mg/ml, respectively. Both enzymes were activated by 1 mM Mn^{2+} and Cu^{2+}, but were completely inhibited by 1 mM Nbromosuccinimide. The GA-II was weakly inhibited by 1 mM p-CMB, dithiothreitol, EDTA, and pyridoxal 5-phosphate, but GA-I was not inhibited by those compounds. Both enzymes had significant ability to digest raw wheat starch and raw rice starch, and hydrolysis rates of raw wheat starch by GA-I and GA-II were 7.8- and 7.3-fold higher than with soluble starch, respectively.

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Characterization of Two Forms of Glucoamylase from Traditional Korean Nuruk Fungi, Aspergillus coreanus NR 15-1 원문보기

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