Immobilization of Cyclodextringlycosyltransferase (CGTase) from Bacillus firmus in Commercial Chitosan
Tóm tắt
The enzyme cyclodextringlycosyltransferase (CGTase) was immobilized in commercial chitosan with different methods of immobilization at different temperatures, with the aim of obtaining a product with improved activity and higher recovery of the free enzyme activity. Three immobilization methods were tested: adsorption, covalent bonding with γ-aminopropyltriethoxysilane (CB-γAPTS), and covalent bonding with hexamethylenediamine (BC-HEMDA). Two test conditions were used, 7 °C without agitation and 26 °C with stirring. The best results were obtained with the method that uses HEMDA as the bifunctional covalent binding agent, giving the highest immobilized enzyme specific activity, 0.263 μmol β-CD/min mg of protein, and highest enzyme activity recovery, 5.2%, when immobilization was carried out at 7 °C.
Tài liệu tham khảo
G. Spagna, R.N. Barbagallo, D. Casarini and P.G. Pifferi: Enzyme and Microb. Technol. 28, 247 (2001).
M. Chellapandian and M.R.V. Krishnan: Proc. Biochem. 33, 595 (1998).
Yang and C.S. Su: J. Chem. Technol. Biotech. 46, 283 (1989).
C.S. Su and C.P. Yang: J. Chem. Technol. Biotech. 48, 313 (1990).
G. Matioli, G.M. Zanin, F. Guimarã es and F.F. de Moraes: Appl. Biochem. Biotech. 70-72, 267 (1998).
E.B. Pereira: Máster Thesis, Universidade Estadual de Maringá, Maringá, PR, Brasil, 128 pp (1999).
P.W. Tardioli, G M. Zanin and F.F. Moares: Appl. Biochem. Biotech. 84-86, 1003 (2000).
M. Dixon and E.C. Webb: Enzymes, 3rd, edn., Longman Group, London, Chap. 2 (1979).
M. Bradford: Anal. Biochem. 72, 248 (1976).
M. Vikmon: in J. Szejtli (ed), The First International Symposium on Cyclodextrin, D. Rildel, Dordrecht, pp. 60–74 (1981).
V. Hamon and F. F. Moraes: in Etude preliminaire a l'immobilsation de l'enzyme CGTase WACKER, Laboratorie de Tecnologie Enzymatique. Université de Tecnologie de Compiè gne (1990).
E. Bon, D.G. Freire, M.F. Mendes and V.F. Soares: Biotech. Bioeng. Symp. 14, 447 (1984).