The purpose of the present work is improving stability properties of porcine pancreatic lipase (triacylglycerol lipase, E.C.220.127.116.11) by immobilization on chitosan/TiO2 composite beads. The immobilization parameters were initial enzyme concentration (0.5-2 mg/ml), adsorption time (5-25 min), and glutaraldehyde concentration (1-4 % v/v). The optimum temperature (20-60 °C), optimum pH (3.0-10.0), kinetic parameters, thermal stability (4-70 °C), pH stability (4.0-9.0), and reusability (9 times) were investigated for characterization of immobilized lipase system. The optimum temperatures of free and immobilized lipase were 30 °C. The temperature profile of the immobilized lipase was spread over a large area. The optimum pH values for the free lipase and immobilized lipase were found to be 6.5 and 7.5, respectively. The thermal stability of immobilized lipase was evaluated, and it maintained 45 % activity at 70 °C. But, at this temperature, soluble lipase protected only 15 % activity. Also, the structural characterization of chitosan/TiO2 composite beads was analyzed with scanning electron microscope (SEM), X-ray diffraction (XRD), thermal gravimetric analysis (TGA), and attenuated total reflection Fourier transform infrared spectroscopy analysis (ATR-FTIR). The significance of this study is improving of stability properties of lipase for the industrial usage especially production of biodiesel and dairy products.
In this study, porcine pancreatic lipase (EC 18.104.22.168) was immobilized on chitin and chitosan by adsorption and subsequent crosslinking with glutaraldehyde, which was added before (conjugation) or after (crosslinking) washing unbound proteins. Conjugation proved to be the better method for both supports. The properties of free and immobilized enzymes were also investigated and compared. The results showed that the pH optimum was shifted from 8.5 to 9.0 for both the immobilized enzymes. Also, the optimum temperature was shifted from 30 to 40 degrees C for chitin-enzyme and to 45 degrees C for chitosan-enzyme conjugates. The immobilization efficiency is low, but the immobilized enzymes have good reusability and stability (storage and operational). Besides these properties, the immobilized lipases were also suitable for catalyzing esterification reactions of fatty acids and fatty alcohols, both with a medium chain length. According to our results, esterification activities of immobilized lipases were two- and four-fold higher for chitosan- and chitin-enzyme, than for the free enzyme, respectively. The immobilization procedure shows a great potential for commercial applications of the immobilized lipase, a relatively low cost commercial enzyme.
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