title: Characterization of an exopolygalacturonase from Aspergillus niger.
abstract: Polygalacturonase (PGI) from Aspergillus niger NRRL3 was purified about 12.0-fold from the cell-free broth using diethylaminoethyl-Sepharose and Sephacryl S-200 columns. The molecular weight of the PGI was 32,000 Da as estimated by gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. PGI had an isoelectric point of 7.6 and an optimum pH of 5.0. PGI was active on polygalacturonic acid and esterified pectins, but the activity on pectin decreased with an increase in degree of esterification. PGI had higher affinity (low Km) and turnover number (Vmax/Km and Kcat/Km) toward polygalacturonic acid. PGI was found to have a temperature optimum at 40 degrees C and was approximately stable up to 30 degrees C. All the examined metal cations had partial inhibitory effects on PGI, while Mn+2 at 5 mM caused a complete inhibition for the enzyme. Comparison of viscosity reduction rates with release of reducing sugars indicated that the enzyme from A. niger is exoacting. The storage stability study of PGI showed that the enzyme in powder form retained 56% of its activity after 9 months of storage at 4 degrees C. The above properties of PGI may be suitable for food processing.
authors.affiliation: Molecular Biology Department, National Research Centre, Cairo, Egypt.
authors.forename: Afaf S
authors.initials: AS
authors.lastname: Fahmy
authors.forename: Fawkia M
authors.initials: FM
authors.lastname: El-Beih
authors.forename: Saleh A
authors.initials: SA
authors.lastname: Mohamed
authors.forename: Somia S
authors.initials: SS
authors.lastname: Abdel-Gany
authors.forename: Engy A
authors.initials: EA
authors.lastname: Abd-Elbaky
mesh_terms: D001234:Aspergillus niger / Q000201:enzymology*; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D004951:Esterification; D006026:Glycoside Hydrolases / Q000378:metabolism*; D007525:Isoelectric Focusing; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D006026:Glycoside Hydrolases; C020136:exopolygalacturonase