title: Two types of phosphorylase from etiolated soybean cotyledons.
abstract: Two types of phosphorylase [EC 2.4.1.1] from the etiolated soybean (Glycine max) cotyledons were separated by column chromatography on DEAE-Sephacel and further purified to apparent homogeneities. Molecular weights of the subunits were 100,000 and 113,000 for phosphorylases I and II, respectively. The native enzymes I and II were a dimer (200,000) and tetramer (450,000), respectively. Electrophoretic analysis by the Hedrick and Smith method indicated that the two phosphorylases were distinct proteins with no correlation. The apparent Km values for glucose 1-phosphate, glycogen, and maltoheptaose of enzyme I were 4.00 mM, 0.18 mg/ml, and 10.3 mM, respectively, while those values of enzyme II were 5.43 mM, 23.8 mg/ml, and 0.30 mM, respectively. The relative activity of enzyme I increased with increasing chain length of the substrate glucans, and waxy maize amylopectin was the best substrate among the 11 saccharides examined. For enzyme II, maltooligosaccharides with degree of polymerization (DP) 5-7 were the better substrates than amylose (DP 38) and glycogen. These results indicated that the soybean phosphorylases I and II have similar properties to a cytoplasmic and chloroplastic type of plant leaf enzyme, respectively.
authors.affiliation: Department of Agricultural Chemistry, Faculty of Agriculture, Tohoku University, Miyagi.
authors.forename: M
authors.initials: M
authors.lastname: Suda
authors.forename: T
authors.initials: T
authors.lastname: Watanabe
authors.forename: M
authors.initials: M
authors.lastname: Kobayashi
authors.forename: K
authors.initials: K
authors.lastname: Matsuda
mesh_terms: D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D005936:Glucans / Q000096:biosynthesis / Q000302:isolation & purification; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008970:Molecular Weight; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism; D006005:Phosphorylases / Q000032:analysis*; D013025:Glycine max / Q000201:enzymology* / Q000378:metabolism; D013696:Temperature
chemical_list: D005936:Glucans; D006005:Phosphorylases; D010744:Phosphoric Monoester Hydrolases