Purification of Commercial Pectinase by Hydrophobic Chromatography

Abstract

A 70-fold purification of a commercial pectinase from Aspergihs niger was achieved by gel filtration, ion exchange and hydrophobic chromatographic columns employed in sequence. An apparent KM of 4.5 x lo-sM and a VM of 1.9 mM/min.mg were obtained. The purified preparation reached a higher degree of purification than those reported in the literature but was not electrophoretically homogeneous. The preparation was an endo-polygalacturonase in nature and preferred high molecular weight pectates as substrates. The pH optimum of 4.4 was not affected by the size of substrates. Hydrophobic chromatography is a useful tool for the preparation of pectic enzymes.

Alkaline thermostable pectinase enzyme from Aspergillus niger strain MCAS2 isolated from Manaslu Conservation Area, Gorkha, Nepal

“…The supernatant was discarded. The precipitate was dissolved in minimum volume of sodium acetate buffer (0.1 M, pH 4.2) and further subjected to Sephadex G-75 (35 × 1.5 cm, bed volume 12–15 ml/g, Sigma company) as per the standard method (Keller et al 2006 ) with certain modifications. The molecular weight and the degree of purity were determined by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDSPAGE) (Sigma, USA) using the wide range protein molecular weight marker (GeNei, India) according to Laemmli ( 1970 ).…”

Production, Purification of Exo-Polygalacturonase from Soil Isolate Paecilomyces variotii NFCCI 1769 and Its Application

“…Carboxymethylcellulose (CMC) was packed into a glass column (15 9 0.55, 10 ml bed volume) and equilibrated with 0.01 mM Tris-HCl buffer (pH 6.0). The concentration gradient of 0.01 to 0.5% w/v of sodium chloride was eluted for elution of protein on a cation exchange resin at a flow rate of 10 ml/h [15]. The fractions were collected and its protein content and enzyme activity were determined as per the method described earlier.…”

“…The isolated strain Paecilomyces variotii NFCCI 1769 particularly showed presence of PG type of pectinase enzyme [15]. The activity of PG at different stages of purification was determined ( Table 2).…”

Molecular Characteristics of Pectins from Enzyme‐Treated Apple Juices

“…These two food grade enzymes contain different concentrations of several enzymes including pectinases, pectin methyl esterases, cellulases, proteases and amylases. The commercial pectic enzyme can be purified if needed (Keller et al, 1982). However, use of purified enzymes in this investigation would not simulate apple juice industry operations.…”