%0 Generic %A D., Fratebianchi %A I.A., Cavello %A S.F., Cavalitto %D 2017 %T Supplementary Material for: Purification and Biochemical and Kinetic Properties of an Endo-Polygalacturonase from the Industrial Fungus Aspergillus sojae %U https://karger.figshare.com/articles/dataset/Supplementary_Material_for_Purification_and_Biochemical_and_Kinetic_Properties_of_an_Endo-Polygalacturonase_from_the_Industrial_Fungus_Aspergillus_sojae/4929530 %R 10.6084/m9.figshare.4929530.v1 %2 https://ndownloader.figshare.com/files/8296754 %2 https://ndownloader.figshare.com/files/8296757 %2 https://ndownloader.figshare.com/files/8296760 %2 https://ndownloader.figshare.com/files/8296763 %K Endo-polygalacturonase %K Enzyme purification %K Biochemical characterization %K Kinetic properties %K Aspergillus sojae %X

An endo-polygalacturonase secreted by Aspergillus sojae was characterized after being purified to homogeneity from submerged cultures with orange peel as the sole carbon source by gel filtration and ion-exchange chromatographies. According to SDS-PAGE and analytical isoelectric focusing analyses, the enzyme presents a molecular weight of 47 kDa and pI value of 4.2. This enzyme exhibits considerable stability under highly acidic to neutral conditions (pH 1.5-6.5) and presents a half-life of 2 h at 50°C. Besides its activity towards pectin and polygalacturonic acid, the enzyme displays pectin-releasing activity, acting best in a pH range of 3.3-5.0. Thin-layer chromatographic analysis revealed that tri-galacturonate is the main enzymatic end product of polygalacturonic acid hydrolysis, indicating that it is an endo-polygalacturonase. The enzyme exhibits Michaelis-Menten kinetics, with KM and VMAX values of 0.134 mg/mL and 9.6 µmol/mg/min, respectively, and remained stable and active in the presence of SO2, ethanol, and various cations assayed except Hg2+.

%I Karger Publishers