Department of Chemistry, Government College Women University, Faisalabad 38040, Pakistan; Environmental & Material Chemistry Laboratory, Department of Chemistry, University of Agriculture, Faisalabad 38040, Pakistan. Electronic address: [Email]
Exo-polygalacturonase (Exo-PG), an extracellular pectinolytic enzyme with great potential in the food industry, was produced from Penicillium janczewskii under solid-state fermentation using wheat bran as a lignocellulosic substrate. Optimization of various fermentation process variables revealed that a moisture level of 70%, d-mannitol (carbon source), yeast extract (nitrogen source) and ZnSO4/NiCl2 as metal salt inducers generated the maximum yield of Exo-PG. After ammonium sulfate precipitation (70% saturation level), the enzyme was characterized in terms of various kinetic (i.e., temperature, pH, inhibitors, Michaelis-Menten constants) and thermodynamic parameters (i.e., Ea, ΔH*, ΔS* and ΔG*). The temperature and pH optima were 45 °C and 6.0, respectively, and the Exo-PG presented stability at a wider range of temperature and pH values. Metal ions Mn+2 markedly improved the enzyme activity, while exposure to urea and ethylenediaminetetraacetic acid drastically inhibited the biocatalytic performance. The kinetic parameters i.e., km and Vmax were computed to be 10 mM and 41.67 U/mL, respectively. A profound increase in clarity, yield, and reduction in viscosity was achieved for different fruit juices (apple, mango, and peach) after treatment with Exo-PG. Total antioxidant and total phenolic contents were also ameliorated. In conclusion, the catalytic activity, thermal steadiness, and fruit juices clarification performance of Exo-PG manifests a great prospect for bio-industrial exploitation.