Abstract
In this paper, solid-phase chemical amination was applied to enrich the surface of Geotrichum candidum lipase (GCL) with amino groups. The aminated enzyme was slightly thermostabilized and showed a different activity profile at different pH in relation to the non-aminated enzyme. Furthermore, the chemical modification allowed the enzyme to be rapid and easily immobilized on carboxymethyl and sulfopropyl agarose-based supports. The cationically exchanged derivatives presented higher thermostability and higher stability at alkaline pH than the soluble aminated enzyme. In addition, the carboxymethyl derivative was 5.4-, 8.7- and 9.1- and the sulfopropyl derivative was 6.6-, 11.7- and 10.7-fold more stable than the free aminated enzyme in methanol, propanol and cyclohexane, respectively. The ionic derivatives were applied for fish oil hydrolysis in organic aqueous medium at 37 °C. After 72 h of reaction, Omega-3 production corresponded to 354 and 482 μM using the carboxymethyl and sulfopropyl derivatives, respectively. These derivatives resulted, respectively, in 2.4- and 3.2-fold increased fish oil hydrolysis when compared to the soluble aminated lipase. After the reaction, carboxymethyl derivative presented 6.65 U per gram of support and sulfopropyl derivative showed 7.85 U per gram of support, making them attractive for industrial application.
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The authors wish to thank CAPES (Brazil) for financial support through the PDSE Program (BEX 14174/13-8) and the “Instituto de Investigación en Ciencia de la Alimentación” (CIAL) for supporting this project.
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de Morais Júnior, W.G., Terrasan, C.R.F., Fernández-Lorente, G. et al. Solid-phase amination of Geotrichum candidum lipase: ionic immobilization, stabilization and fish oil hydrolysis for the production of Omega-3 polyunsaturated fatty acids. Eur Food Res Technol 243, 1375–1384 (2017). https://doi.org/10.1007/s00217-017-2848-8
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DOI: https://doi.org/10.1007/s00217-017-2848-8