10.6084/m9.figshare.4929530.v1 Fratebianchi D. Fratebianchi D. Cavello I.A. Cavello I.A. Cavalitto S.F. Cavalitto S.F. Supplementary Material for: Purification and Biochemical and Kinetic Properties of an Endo-Polygalacturonase from the Industrial Fungus Aspergillus sojae Karger Publishers 2017 Endo-polygalacturonase Enzyme purification Biochemical characterization Kinetic properties Aspergillus sojae 2017-04-28 07:05:50 Dataset 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 <p>An endo-polygalacturonase secreted by <i>Aspergillus sojae </i>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 K<sub>M</sub> and V<sub>MAX</sub> values of 0.134 mg/mL and 9.6 µmol/mg/min, respectively, and remained stable and active in the presence of SO<sub>2</sub>, ethanol, and various cations assayed except Hg<sup>2+</sup>.</p>