Biochemical Characterization of Two Polygalacturonases Purified from the Digestive Juice of the Snail Limicolaria flammea

Abstract

Polygalacturonases constitute the major part of pectinase preparations for many bioprocess purposes. Investigation on the digestive juice of snail Limicolaria flammea led to purification of two polygalacturonases named PG1 and PG2. Properties of these enzymes were examined to explore their potential in biotechnology applications. A three steps procedure including size exclusion, anion and cation exchange and hydrophobic interaction chromatography were used for purification. The enzymes PG1 and PG2 had native molecular weights of approximately 46 and 86 kDa, respectively and functioned both as monomeric structures. The purified polygalacturonases PG1 and PG2 showed optimum hydrolysis activities at 50°C in sodium acetate buffer pH 5.6. The common inhibitor of the two purified polygalacturonases activity were Mn2+, Ca2+, Zn2+, EDTA, SDS and L-cystein. NH3+ stimulate the polygalacturonase PG1 while Ba2+ was an activator for polygalacturonase PG2. Substrate specificity indicated that these enzymes hydrolyse a broad range of pectin from different sources. The highest activity of PG1 was observed with apple pectin and lemon pectin while PG2 showed its highest activity with orange pectin. The catalytic efficiency of PG1 was highest for lemon pectin (0.125 µmol/min/mL) and orange pectin (0.124 µmol/min/mL). PG2 displayed highest catalytic efficiency (0.325 µmol/min/mL) towards orange pectin. These results suggest that orange and lemon pectin would be the potential physiological substrates of the two purified enzymes.

Kinetic and Thermodynamic Characterization of two Polygalacturonases Isolated from the Digestive Juice of the Snail Limicolaria flammea

“…To our knowledge, very few studies have been carried out to polygalacturonases animal sources. As regards polygalacturonases PG1 and PG2 previously isolated from the digestive juice of the snail Limicolaria flammea by Kanga et al [20], no study has been devoted to their thermal stability parameters. Therefore, objective of the current study was to determined thermal stability parameters (Speed consistency, activation energy, half-life time, decimal reduction time, temperature of destruction, temperature coefficient, enthalpy, entropy et Gibbs free energy) of the two polygalacturonases purified from digestive juice of the snail Limicolaria flammea [20] for several industrial applications such as fruit juice clarification.…”