Beta-amylase in germinating millet seeds.
Beta-amylase (EC 3.2.1.2) was isolated from germinating millet (Panicum miliaceum L.) seeds by a procedure that included ammonium sulfate fractionation, chromatography on DEAE-cellulofine and CM-cellulofine, and preparative isoelectric focusing. The enzyme was homogeneous by SDS-PAGE. The M(r) of the enzyme was estimated to be 58,000 based on its mobility on SDS-PAGE and gel filtration with TSKgel G4000SW(XL), which showed that it is composed of a single unit. The isoelectric point of the enzyme was 4.62. The enzyme hydrolyzed malto-oligosaccharides more readily as their degree of polymerization increased, this being strongest for malto-oligosaccharides larger than 13 glucose residues and very weakly for maltotriose. Amylose, amylopectin and soluble starch were the most suitable substrates for the enzyme. While the enzyme showed some activity against native starch by itself, starch digestion was accelerated 2.5-fold using alpha-amylase, pullulanase and alpha-glucosidase. This enzyme appears to be very important for the germination of millet seeds.
Yamasaki
Yoshiki
Y
Research Institute for Bioresources, Okayama University, Kurashiki-shi, Okayama 710-0046, Japan. yosikiy@rib.okayama-u.ac.jp
D018525:Germination; D006026:Glycoside Hydrolases / Q000378:metabolism; D007526:Isoelectric Point; D008970:Molecular Weight; D009844:Oligosaccharides / Q000737:chemistry / Q000378:metabolism; D008897:Panicum / Q000201:enzymology* / Q000254:growth & development*; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D012639:Seeds / Q000201:enzymology; D013213:Starch / Q000031:analogs & derivatives / Q000032:analysis / Q000378:metabolism; D013379:Substrate Specificity; D000520:alpha-Glucosidases / Q000378:metabolism; D001614:beta-Amylase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*
D009844:Oligosaccharides; D010940:Plant Proteins; C021705:maltooligosaccharides; D013213:Starch; D006026:Glycoside Hydrolases; D001614:beta-Amylase; D000520:alpha-Glucosidases; C100181:pullulanase