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>