Tannases (E.C. 22.214.171.124) are esterases who hydrolyse the "ester" bond and the "depside" bond in substrates such as tannic acid, epicatechin gallate, epigallocatechin gallate in aqueous medium and synthesize esters of gallic acid in medium containing alcool and organic solvent. Enzymes are used in various industrial sectors, however, their use often is limited due to its low stability and high cost, one way to solve this problem is through the enzyme immobilization. The synthesis of gallates is important for producing natural compounds with high antioxidant capacity, which can be used in place of those as synthetic BHA, BHT and TBHQ. This project aims to study and optimize the immobilization of the enzyme tannase by Paecilomyces variotti in different suports. Than free and immobilized enzyme will be used to study the synthesis of esters of gallic acid and the hydrolisis of pure (commercial) galic and elagic phenolic compounds . The reactions of synthesis of esters will be optimized using experimental design, the variables under study for the synthesis will be concentration of substrate and enzyme, temperature, agitation, pH, concentration of water and sorbitol and for hydrolysis concentration of substrate and enzyme, temperature, agitation and pH. The reaction will be monitored after 2, 4, 6, 12 and 24 hours of the start of the experiment.
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