RESUMO
Immobilization is practical to upgrade enzymes, increasing their performance and expanding their applications. The recombinant, solvent tolerant lipase LipA PSA01 from Pseudomonas aeruginosa was immobilized on polypropylene Accurel® MP1004 to improve its performance. We investigated the effect of ethanol as an additive during the immobilization process at three concentrations (20%, 25%, and 30%) on the operational behavior of the enzyme. The immobilization efficiency was higher than 92%, and the immobilized enzymes showed hyperactivation and thermal resistance depending on the concentration of ethanol. For example, at 70 °C, the free enzyme lost the activity, while the prepared one with ethanol 25% conserved a residual activity of up to 73.3% (∆ T15 50 = 27.1 °C). LipA immobilized had an optimal pH value lower than that of the free enzyme, and the organic solvent tolerance of the immobilized enzymes depended on the ethanol used. Hence, the immobilized enzyme with ethanol 25% showed hyperactivation to more solvents than the soluble enzyme. Remarkable stability towards methanol (up to 8 folds) was evidenced in all the immobilized preparations. The immobilized enzyme changed their chemo preference, and it hydrolyzed oils preferentially with short-chain than those with long-chain. LipA had a notable shelf-life after one year, keeping its activity up to 87%. Ethanol facilitated the access of the enzyme to the hydrophobic support and increased its activity and stability according to the amount of ethanol added.
