Sonohydrolysis using an enzymatic cocktail in the preparation of free fatty acid.

In this work, the concept of lipase cocktail has been proposed in the ultrasound-assisted hydrolysis of coconut oil. Lipase from Thermomyces lanuginosus (TLL), lipase from Rhizomucor miehei (RML), and lipase B from Candida antarctica (CALB) were evaluated as biocatalysts in different combinations. The best conversion (33.66%) was achieved using only RML; however, the best lipase cocktail (75% RML and 25% CALB) proposed by the triangular response surface was used to achieve higher conversions. At the best lipase cocktail, reaction parameters [temperature, biocatalyst content and molar ratio (water/oil)] were optimized by a Central Composite Design, allowing to obtain more than 98% of conversion in the hydrolysis of coconut oil in 3 h of incubation at 37 kHz, 300 W and 45 °C by using 20% of the lipase cocktail (w/w) and a molar ratio of 7.5:1 (water/oil). The lipase cocktail retained about 50% of its initial activity after three consecutive cycles of hydrolysis. To the authors' knowledge, up to date, this communication is the first report in the literature for the ultrasound-assisted hydrolysis of coconut oil catalyzed by a cocktail of lipases. Under ultrasound irradiation, the concept of lipase cocktail was successfully applied, and this strategy could be useful for the other types of reactions using heterogeneous substrates.

Ethyl Butyrate Synthesis Catalyzed by Lipases A and B from Candida antarctica Immobilized onto Magnetic Nanoparticles. Improvement of Biocatalysts' Performance under Ultrasonic Irradiation.

The synthesis of ethyl butyrate catalyzed by lipases A (CALA) or B (CALB) from Candida antarctica immobilized onto magnetic nanoparticles (MNP), CALA-MNP and CALB-MNP, respectively, is hereby reported. MNPs were prepared by co-precipitation, functionalized with 3-aminopropyltriethoxysilane, activated with glutaraldehyde, and then used as support to immobilize either CALA or CALB (immobilization yield: 100 ± 1.2% and 57.6 ± 3.8%; biocatalysts activities: 198.3 ± 2.7 Up-NPB/g and 52.9 ± 1.7 Up-NPB/g for CALA-MNP and CALB-MNP, respectively). X-ray diffraction and Raman spectroscopy analysis indicated the production of a magnetic nanomaterial with a diameter of 13.0 nm, whereas Fourier-transform infrared spectroscopy indicated functionalization, activation and enzyme immobilization. To determine the optimum conditions for the synthesis, a four-variable Central Composite Design (CCD) (biocatalyst content, molar ratio, temperature and time) was performed. Under optimized conditions (1:1, 45 °C and 6 h), it was possible to achieve 99.2 ± 0.3% of conversion for CALA-MNP (10 mg) and 97.5 ± 0.8% for CALB-MNP (12.5 mg), which retained approximately 80% of their activity after 10 consecutive cycles of esterification. Under ultrasonic irradiation, similar conversions were achieved but at 4 h of incubation, demonstrating the efficiency of ultrasound technology in the enzymatic synthesis of esters.

Immobilization of Lipase A from Candida antarctica onto Chitosan-Coated Magnetic Nanoparticles.

In this communication, lipase A from Candida antarctica (CALA) was immobilized by covalent bonding on magnetic nanoparticles coated with chitosan and activated with glutaraldehyde, labelled CALA-MNP, (immobilization parameters: 84.1% ± 1.0 for immobilization yield and 208.0 ± 3.0 U/g ± 1.1 for derivative activity). CALA-MNP biocatalyst was characterized by X-ray Powder Diffraction (XRPD), Fourier Transform Infrared (FTIR) spectroscopy, Thermogravimetry (TG) and Scanning Electron Microscope (SEM), proving the incorporation of magnetite and the immobilization of CALA in the chitosan matrix. Besides, the immobilized biocatalyst showed a half-life 8-11 times higher than that of the soluble enzyme at pH 5-9. CALA showed the highest activity at pH 7, while CALA-MNP presented the highest activity at pH 10. The immobilized enzyme was more active than the free enzyme at all studied pH values, except pH 7.