caa a22 4500 606212272 CHVBK 20210128101042.0 cr unu---uuuuu 210128e20150701xx s 000 0 eng 10.1007/s10295-015-1619-4 doi (NATIONALLICENCE)springer-10.1007/s10295-015-1619-4 An efficient method for the immobilization of inulinase using new types of polymers containing epoxy groups [Elektronische Daten] [Mariusz Trytek, Jan Fiedurek, Beata Podkościelna, Barbara Gawdzik, Marcin Skowronek] New glycidyl methacrylate copolymers containing different numbers of epoxy groups were synthesized and used to develop effective procedures for inulinase immobilization. The beneficial characteristics of the carriers included a high degree of crosslinking, stability at ambient temperature, an appropriate surface, and the presence of reactive epoxy groups. Some factors affecting the efficiency of immobilization of crude inulinase, including the kind and amount of carrier, the number of epoxy groups, as well as buffer pH and buffer concentration were examined. The yield of immobilization of this enzyme on the investigated type of microspheres was higher than on the commercial carrier, Eupergit® C. After immobilization, the optimum temperature for inulinase activity shifted from 55 to 45°C, whereas the optimum pH=5 remained unchanged. The basic parameters of inulin hydrolysis were examined, and the possibility of applying the obtained biocatalyst in continuous conditions was tested. Inulin at a concentration of 0.5% (w/v) was almost completely hydrolyzed to fructose (in a yield of 98%) at a flow rate of 0.1mL/min. A tenfold increase in the speed of flow resulted in an increase in the yield of oligosaccharides (DP2-DP6) up to ~41% in the overall hydrolysate, as analysed by HPLC-RID and LC-ESI/MS. These results indicate that two forms of inulinase, an exo- and an endo-acting enzyme, were immobilized on our carrier. The enzyme showed good operational stability in a packed column over 28days. There were no significant decreases in the efficiency of continuous hydrolysis during this time (about 17.4% in comparison to its initial value). The Author(s), 2015 Copolymers nationallicence Epoxy groups nationallicence Eupergit nationallicence Immobilization nationallicence Inulinase nationallicence Trytek Mariusz Department of Industrial Microbiology, Institute of Microbiology and Biotechnology, Faculty of Biology and Biotechnology, Maria Curie-Skłodowska University, Akademicka St. 19, 20-033, Lublin, Poland aut Fiedurek Jan Department of Industrial Microbiology, Institute of Microbiology and Biotechnology, Faculty of Biology and Biotechnology, Maria Curie-Skłodowska University, Akademicka St. 19, 20-033, Lublin, Poland aut Podkościelna Beata Department of Polymer Chemistry, Faculty of Chemistry, Maria Curie-Sklodowska University, Maria Curie-Skłodowska Sq. 5, 20-031, Lublin, Poland aut Gawdzik Barbara Department of Polymer Chemistry, Faculty of Chemistry, Maria Curie-Sklodowska University, Maria Curie-Skłodowska Sq. 5, 20-031, Lublin, Poland aut Skowronek Marcin Department of Industrial Microbiology, Institute of Microbiology and Biotechnology, Faculty of Biology and Biotechnology, Maria Curie-Skłodowska University, Akademicka St. 19, 20-033, Lublin, Poland aut Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/7(2015-07-01), 985-996 1367-5435 42:7<985 2015 42 10295 https://doi.org/10.1007/s10295-015-1619-4 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s10295-015-1619-4 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Trytek Mariusz Department of Industrial Microbiology, Institute of Microbiology and Biotechnology, Faculty of Biology and Biotechnology, Maria Curie-Skłodowska University, Akademicka St. 19, 20-033, Lublin, Poland aut NATIONALLICENCE 700 1- Fiedurek Jan Department of Industrial Microbiology, Institute of Microbiology and Biotechnology, Faculty of Biology and Biotechnology, Maria Curie-Skłodowska University, Akademicka St. 19, 20-033, Lublin, Poland aut NATIONALLICENCE 700 1- Podkościelna Beata Department of Polymer Chemistry, Faculty of Chemistry, Maria Curie-Sklodowska University, Maria Curie-Skłodowska Sq. 5, 20-031, Lublin, Poland aut NATIONALLICENCE 700 1- Gawdzik Barbara Department of Polymer Chemistry, Faculty of Chemistry, Maria Curie-Sklodowska University, Maria Curie-Skłodowska Sq. 5, 20-031, Lublin, Poland aut NATIONALLICENCE 700 1- Skowronek Marcin Department of Industrial Microbiology, Institute of Microbiology and Biotechnology, Faculty of Biology and Biotechnology, Maria Curie-Skłodowska University, Akademicka St. 19, 20-033, Lublin, Poland aut NATIONALLICENCE 773 0- Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/7(2015-07-01), 985-996 1367-5435 42:7<985 2015 42 10295