Cyclodextrin production by Bacillus lehensis isolated from cassava starch: Characterisation of a novel enzyme
Blanco, K.C., University Estadual Paulista, Rio Claro (Brazil). Dept. of Biochemistry and Microbiology | De Moraes, F.F., University Estadual de Maringa (Brazil). Chemical Engineering Dept. | Bernardi, N.S., University Estadual Paulista, Rio Claro (Brazil). Dept. of Biochemistry and Microbiology | Vettori, M.H.P.B., University Estadual Paulista, Rio Claro (Brazil). Dept. of Biochemistry and Microbiology | Monti, R., University Estadual Paulista, Rio Claro (Brazil). Dept. of Biochemistry and Microbiology | Contiero, J., University Estadual Paulista, Rio Claro (Brazil). Dept. of Biochemistry and Microbiology
The properties of a previously unknown enzyme, denominated cyclodextrin glycosyltransferase, produced from Bacillus lehensis, were evaluated using affinity chromatography for protein purification. Enzyme characteristics (optimum pH and temperature; pH and temperature stability), the influence of substances on the enzyme activity, enzyme kinetics, and cyclodextrin production were analysed. Cyclodextrin glycosyltransferase was purified up to 320.74-fold by affinity chromatography using beta-cyclodextrin as the binder and it exhibited 8.71% activity recovery. This enzyme is a monomer with a molecular weight of 81.27 kDa, as estimated by SDS-PAGE. Optimum temperature and pH for cyclodextrin glycosyltransferase were 55 deg C and 8.0, respectively. The Michaelis-Menten constant was 8.62 g/l during maximum velocity of 0.858 g/l/h.
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