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In bioprocesses lipases are typically used in immobilized form, irrespective of type of reaction systems, to ensure an even distribution of catalysts in water restricted media and/or to facilitate separation and reuse. In these studies we report on the selection of appropriate enzyme-carrier preparation for hydrolysis reaction in aqueous and biphasic systems and transesterification in organic solvent. For this Candida rugosa lipase was bound by adsorption or covalent attachment onto various carriers to give 24 preparations. Selection of proper preparation was based on reactivity, thermal stability (4 h at 60°C), possibility of drying and operational stability in 17-23 successive batch processes of 4-nitrophenyl palmitate hydrolysis in water. Activity of preparations varied from 20 to 5100 U∙mL-1 but the most stable preparations were those of moderate activity: bound by adsorption or covalent attachment to NH2-Kieselgel or acrylic carrier (retained activity over 90%). Selected preparations were used for hydrolysis of ethyl (1-butyryloxyethyl)-phenylphosphinate in biphasic system, and, after drying, in ethyl (1-hydroxyethyl)-phenyl-phosphinate transesterification. In this study operational stability was the principal criterion of selection. In water system, lipase covalently bound to NH2-Kieselgel was the best - preserved 50% of initial activity in consecutive batch processes. In biphasic system and lipase covalently bound to acrylic and NH2-Kieselgel the values were 90 or 77%, respectively, whereas in organic solvent, when lipase was immobilized on NH2-Kieselgel by adsorption, it was 50%. Thus, NH2-Kieselgel appears to be an universal matrix for investigated lipase immobilization and can be used in all reaction systems.
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p.1-6,fig.,ref.
Twórcy
autor
- Wroclaw University of Technology, Faculty of Chemistry, Department of Bioorganic Chemistry, Poland
autor
- Wroclaw University of Technology, Faculty of Chemistry, Department of Bioorganic Chemistry, Poland
autor
- Wroclaw University of Technology, Faculty of Chemistry, Department of Bioorganic Chemistry, Poland
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- Silesian University of Technology, Department of Chemical Engineering, Poland
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- Kaunas University of Technology, Faculty of Chemical Technology, Lithuania
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- Silesian University of Technology, Department of Chemical Engineering, Poland
- Polish Academy of Sciences, Institute of Chemical Engineering, Poland
autor
- Wroclaw University of Technology, Faculty of Chemistry, Department of Bioorganic Chemistry, Poland
Bibliografia
- Adlercreutz P (2013) Immobilisation and application of lipases in organic media. Chem Soc Rev 42: 6406-6436.
- Bryjak J, Aniulyte J, Liesiene J (2007) Evaluation of mantailored cellulose-based carriers in glucoamylase immobilization. Carbohyd Res 342: 1105-1109.
- Domínguez de MP, Sánchez-Montero JM, Sinisterra JV, Alcántara AR (2006) Understanding Candida rugosa lipases: An overview. Biotechnol Adv 24: 180-196.
- Jarzębski AB, Szymańska K, Bryjak J, Mrowiec-Białoń J (2007) Covalent immobilization of trypsin on to siliceous mesostructured cellular foams to obtain effective biocatalysts. Catal Today 124: 2-10.
- Knežević ZD, Šiler-Marinković SS, Mojović LV (2004) Immobilized lipases as practical catalysts. APTEFF 35: 151-164.
- Liese A, Seelbach K, Buchholz A, Haberland J (2006) Processes. In Industrial Biotransformation, 2nd edn. Liese A, Seelbach K, Wandrey Ch, eds, pp 273-320. Wiley-VCH, Weinheim.
- Lowry OH, Rosebrough N J, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265-275.
- Majewska P, Kafarski P, Lejczak B (2006) Simple and effective method for the deracemization of ethyl 1-hydroxyphosphinate using biocatalysts with lipolytic activity. Tetrahedron Asymmetr 17: 2870-2875.
- Pernas MA, Pastrana L, Fuciños P, Rúa ML (2009) Regulation of the interfacial activation within the Candida rugosa lipase family. J Phys Org Chem 22: 508-514.
- Rekuć A, Bryjak J, Szymańska K, Jarzębski AB (2009) Laccase immobilization on mesostructured cellular foams affords preparations with ultra high activity. Process Biochem 44: 191-198.
- Schmid RD, Verger R (1998) Lipases: Interfacial enzymes with attractive applications. Angew Chem Int Ed Engl 37: 1608-1633.
- Szymańska K, Bryjak J, Mrowiec-Białoń J, Jarzębski AB (2007) Application and properties of siliceous mesostructured cellular foams as enzymes carriers to obtain efficient biocatalysts. Micropor Mesopor Mat 99: 167-175.
- Zynek K, Bryjak J, Szymańska K, Jarzębski AB (2011) Screening of porous and cellular materials for covalent immobilisation of Agaricus bisporus tyrosinase. Biotechnol Bioproc Eng 16: 180-189.
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Bibliografia
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