Production, purification and characterization of the exo-polygalacturonase (pectinase) produced by the thermophilic fungus Rhizomucor pusillus, in submerged fermentation

• Grant number: 12/01198-6
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: April 01, 2012
• End date: December 31, 2012
• Field of knowledge: Biological Sciences - Biochemistry - Enzymology
• Principal Investigator: Gustavo Orlando Bonilla Rodriguez
• Grantee: Lucas Vinícius Trindade
• Host Institution: Instituto de Biociências, Letras e Ciências Exatas (IBILCE). Universidade Estadual Paulista (UNESP). Campus de São José do Rio Preto. São José do Rio Preto , SP, Brazil

Abstract

The temperature is one of main factors that affect the microorganisms' growth, including the fungi, which can be distinguished from each other by the maximum and minimum temperature of growth. Generally, the thermophilic fungi have a minimum temperature of growth around 20°C and a maximum temperature of growth around 50°C. These fungi produce differentiated bioproducts, like enzymes. The present study aims to produce the exo-polygalacturonase (pectinase) of the thermophilic fungus Rhizomucor pusillus, purify it and characterize it. This project is motivated by the interesting properties of enzymes from thermophilic fungi, which have thermal stability properties, and also due to the wide application of pectinases in the industry. The fungus Rhizomucor pusillus will be cultivated in submerged fermentation (SmF). The crude enzyme solution will be submitted to an enzymatic assay and after it will be purified by gel filtration and ion-exchange chromatography. Techniques such as SDS-PAGE and isoelectric focusing will be performed and the enzyme will be characterized as to pH and temperature optimum of activity, thermal stability, stability related to the pH, specific substrate, and effect of ions on enzyme activity. So it will be possible to determine the kinetic and thermodynamic parameters of exo-polygalacturonase. (AU)