A simple, rapid, and economic method of enzyme immobilization was developed for phospholipase Lecitase® ultra (LU) via interfacial adsorption. The effect of nature of the polystyrene supports and the kinetic behavior and stability of immobilized lecitase® ultra (IM-LU) were evaluated. Six macroporous resins (AB-8, X-5, DA-201, NKA-9, D101, D4006) and two anion resins (D318 and D201) were studied as the supports. DA-201 resin was selected because of its best immobilization effect for LU. Immobilization conditions were investigated, including immobilization time, pH, and enzyme concentration. IM-LU with a lipase activity of 1,652.4 ± 8.6 U/g was obtained. The adsorption process was modeled by Langmuir and Freundlich equations, and the experimental data were better fit for the former one. The kinetic constant (K m) values were found to be 192.7 ± 2.2 mM for the free LU and 249.3 ± 5.4 mM for the IM-LU, respectively. The V max value of free LU (169.5 ± 4.3 mM/min) was higher than that of the IM-LU (53.8 ± 1.5 mM/min). Combined strategies of scanning electron micrograph, thermogravimetric analysis, and Fourier transform infrared (FTIR) spectroscopy were employed to characterize the IM-LU. FTIR spectroscopy showed that the secondary conformation of the enzyme had changed after immobilization, through which a decrease of α-helix content and an increase of β-sheet content were observed. The IM-LU possessed an improved thermal stability as well as metal ionic tolerance when compared with its free form. The reusability of IM-LU was also evaluated through catalyzing esterification reaction between oleic acid and glycerol. It exhibited approximately 70 % of relative esterification efficiency after six successive cycles. This immobilized enzyme on hydrophobic support may well be used for the synthesis of structural lipids in lipid area.
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Immobilized Lecitase® ultra
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The financial support from the National Natural Science Foundation of China under grant 31000793, the Ministry of Science and Technology of People’s Republic of China under grant 2011BAD02B04, and the Science and Technology Council of Guangdong under grants 2009A020700003 and 2009B080701063 are gratefully acknowledged.
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Liu, N., Fu, M., Wang, Y. et al. Immobilization of Lecitase® Ultra onto a Novel Polystyrene DA-201 Resin: Characterization and Biochemical Properties. Appl Biochem Biotechnol 168, 1108–1120 (2012). https://doi.org/10.1007/s12010-012-9845-y
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