title: Purification and properties of two forms of glucoamylase from Aspergillus niger.
abstract: A. niger produced alpha-glucosidase, alpha-amylase and two forms of glucoamylase when grown in a liquid medium containing raw tapioca starch as the carbon source. The glucoamylases, which formed the dominant components of amylolytic activity manifested by the organism, were purified to homogeneity by ammonium sulfate precipitation, ion-exchange and two cycles of gel filtration chromatography. The purified enzymes, designated GA1 and GA2, a raw starch digesting glucoamylase, were found to have molar masses of 74 and 96 kDa and isoelectric points of 3.8 and 3.95, respectively. The enzymes were found to have pH optimum of 4.2 and 4.5 for GA1 and GA2, respectively, and were both stable in a pH range of 3.5-9.0. Both enzymes were thermophilic in nature with temperature optimum of 60 and 65 degrees C, respectively, and were stable for 1 h at temperatures of up to 60 degrees C. The kinetic parameters Km and V showed that with both enzymes the branched substrates, starch and amylopectin, were more efficiently hydrolyzed compared to amylose. GA2, the more active of the two glucoamylases produced, was approximately six to thirteen times more active towards raw starches compared to GA1.
authors.affiliation: School of Biological Sciences, Universiti Sains Malaysia, Penang, Malaysia.
authors.forename: A A
authors.initials: AA
authors.lastname: Amirul
authors.forename: S L
authors.initials: SL
authors.lastname: Khoo
authors.forename: M N
authors.initials: MN
authors.lastname: Nazalan
authors.forename: M S
authors.initials: MS
authors.lastname: Razip
authors.forename: M N
authors.initials: MN
authors.lastname: Azizan
mesh_terms: D000687:Amylopectin / Q000378:metabolism; D000688:Amylose / Q000378:metabolism; D001234:Aspergillus niger / Q000201:enzymology*; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D005087:Glucan 1,4-alpha-Glucosidase / Q000302:isolation & purification* / Q000378:metabolism*; D006361:Heating; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D007526:Isoelectric Point; D007700:Kinetics; D013213:Starch / Q000378:metabolism
chemical_list: D013213:Starch; D000688:Amylose; D000687:Amylopectin; D005087:Glucan 1,4-alpha-Glucosidase