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Pectin hydrolysis in cashew apple juice by Aspergillus aculeatus URM4953 polygalacturonase covalently-immobilized on calcium alginate beads: A kinetic and thermodynamic study

Author:
Silva, Jônatas de Carvalho, de França, Pedro Renann Lopes, Converti, Attilio, Porto, Tatiana Souza
Source:
International journal of biological macromolecules 2019 v.126 pp. 820-827
ISSN:
0141-8130
Subject:
Aspergillus aculeatus, Gibbs free energy, activation energy, calcium alginate, enthalpy, entropy, equations, hydrolysis, pectins, polygalacturonase, protein unfolding, temperature
Abstract:
The kinetics and thermodynamics of pectin hydrolysis in cashew apple juice by polygalacturonase (PG) from Aspergillus aculeatus URM4953 covalently-immobilized on calcium alginate beads were investigated. Immobilized-PG activity in cashew apple juice was the highest at 20 °C, showing a maximum hydrolysis rate of 58.2 mg/mL·min, a catalytic constant of 166.2 s−1 and an affinity constant of 113.0 mg/mL. Since the enzyme exhibited an allosteric behavior, the hydrolysis rate was modeled, with excellent accuracy, by the Hill Equation as function of pectin concentration. The Hill coefficient increased from 3 to 5 with increasing temperature from 20 to 50 °C, evidencing a positive cooperativity mechanism. The reaction activation energy and the standard enthalpy variation of enzyme unfolding were 80.3 and 16.6 kJ/mol, respectively. Consistently with the kinetic results, PG-catalyzed pectin hydrolysis proceeded with maximum spontaneity at 20 °C, showing activation Gibbs free energy, enthalpy and entropy of 59.3 kJ/mol, 77.9 kJ/mol and 63.4 J/mol·K, respectively. Immobilized PG was successful in the hydrolysis of cashew apple juice pectin, requiring a low temperature to act optimally.
Agid:
6290767