Sucrose hydrolysis by gelatin-immobilized inulinase from Kluyveromyces marxianus var. bulgaricus |
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Authors: | Fabrí cio C. de Paula,Má rcia Luciana Cazetta,Rubens Monti,Jonas Contiero |
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Affiliation: | 1. Department of Biochemistry and Microbiology, Biological Sciences Institute, São Paulo State University-Unesp, Av. 24A 1515, Rio Claro 13506900, Brazil;2. Department of Food and Nutrition, Faculty of Pharmaceutical Sciences, São Paulo State University-Unesp, Brazil |
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Abstract: | The crude cell-free medium from a culture of Kluyveromyces marxianus var. bulgaricus was immobilized in a gelatin-water support, with an immobilization yield of 82.60% for inulinase activity. The optimum pH for both free and immobilized inulinase was the same (3.5) and the optimum temperatures were 55 °C for the free and 60 °C for the immobilized enzyme. The Arrhenius plots were linear and activation energies were 56.20 (free enzyme) and 20.27 kJ/mol K (immobilized enzyme). The kinetic parameters were calculated by Lineweaver–Burk plots and the Vmax and Km were 37.60 IU/mg protein and 61.83 mM for the free inulinase and 31.45 IU/mg protein and 149.28 mM for the immobilized enzyme, respectively. The operational stability of the immobilized inulinase was studied in a continuous fixed-bed column reactor for 33 days, at the end of which the sucrose conversion was 58.12%. |
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Keywords: | Immobilization Inulinase Inverted sugar Kluyveromyces marxianus var. bulgaricus |
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