title: Biosynthesis of glycogen in Neurospora crassa. Purification and properties of the branching enzyme.
abstract: A branching enzyme was extracted from the mycelia of Neurospora crassa and was purified to electrophoretic homogeneity by procedures including DEAE-Sephacel column chromatography, 6-aminohexyl-Sepharose 4B column chromatography and gel filtration on Toyopearl HW-55S. The final yield of the branching enzyme activity was 15.1%, and the final purified enzyme preparation showed a specific activity of 702 units per mg of protein. The molecular weight of this enzyme was estimated to be 80,000 by electrophoresis in sodium dodecyl sulfate-polyacrylamide gel. The amino acid composition and the carbohydrate content of this enzyme were analyzed. The isoelectric point of this enzyme determined by polyacrylamide gel isoelectrofocusing was 5.6. The branching activity of the enzyme was confirmed by its action on amylopectin as well as by the combined action of this enzyme and N. crassa glycogen synthase. The action of this enzyme on amylopectin decreased the wavelength of the absorption maximum of the glucan-iodine complex, and increased the amount of the short unit chains of the debranched product. The product obtained by the combined action yielded beta-limit dextrin upon hydrolysis with beta-amylase. No multiplicity was found for the branching activity either by chromatography or by electrophoresis.
mesh_terms: D015061:1,4-alpha-Glucan Branching Enzyme / Q000302:isolation & purification*; D000596:Amino Acids / Q000302:isolation & purification; D055598:Chemical Phenomena; D002621:Chemistry; D002845:Chromatography / Q000379:methods; D004589:Electrophoresis, Disc; D005964:Glucosyltransferases / Q000302:isolation & purification*; D006003:Glycogen / Q000096:biosynthesis*; D007525:Isoelectric Focusing; D009491:Neurospora / Q000201:enzymology*; D009492:Neurospora crassa / Q000201:enzymology*
chemical_list: D000596:Amino Acids; D006003:Glycogen; D005964:Glucosyltransferases; D015061:1,4-alpha-Glucan Branching Enzyme