title: Purification and characterization of an alpha-glucosidase from germinating millet seeds.
abstract: An alpha-glucosidase (alpha-D-glucoside glucohydrolase, EC 3.2.1.20) was isolated from germinating millet (Panicum miliaceum L.) seeds by a procedure that included ammonium sulfate fractionation, chromatography on CM-cellulofine/Fractogel EMD SO(3), Sephacryl S-200 HR and TSK gel Phenyl-5 PW, and preparative isoelectric focusing. The enzyme was homogenous by SDS-PAGE. The molecular weight of the enzyme was estimated to be 86,000 based on its mobility in SDS-PAGE and 80,000 based on gel filtration with TSKgel super SW 3000, which showed that it was composed of a single unit. The isoelectric point of the enzyme was 8.3. The enzyme readily hydrolyzed maltose, malto-oligosaccharides, and alpha-1,4-glucan, but hydrolyzed polysaccharides more rapidly than maltose. The K(m) value decreased with an increase in the molecular weight of the substrate. The value for maltoheptaose was about 4-fold lower than that for maltose. The enzyme preferably hydrolyzed amylopectin in starch, but also readily hydrolyzed nigerose, which has an alpha-1,3-glucosidic linkage and exists as an abnormal linkage in the structure of starch. In particular, the enzyme readily hydrolyzed millet starch from germinating seeds that had been degraded to some extent.
authors.affiliation: Research Institute for Bioresources, Okayama University, Kurashiki-shi, Okayama 710-0046, Japan. yosikiy@rib.okayama-u.ac.jp
authors.forename: Yoshiki
authors.initials: Y
authors.lastname: Yamasaki
authors.forename: Mikio
authors.initials: M
authors.lastname: Fujimoto
authors.forename: Junji
authors.initials: J
authors.lastname: Kariya
authors.forename: Haruyoshi
authors.initials: H
authors.lastname: Konno
mesh_terms: D018525:Germination; D005936:Glucans / Q000378:metabolism; D008897:Panicum / Q000201:enzymology*; D012639:Seeds / Q000201:enzymology*; D013379:Substrate Specificity; D013997:Time Factors; D000520:alpha-Glucosidases / Q000302:isolation & purification* / Q000378:metabolism*
chemical_list: D005936:Glucans; D000520:alpha-Glucosidases