Cyclodextrin production by Bacillus lehensis isolated from cassava starch: Characterisation of a novel enzyme
K.C. Blanco, F.F. de Moraes, N.S. Bernardi, M.H.P.B. Vettori, R. Monti, J. Contierohttps://doi.org/10.17221/432/2012-CJFSCitation:Blanco K.C., de Moraes F.F., Bernardi N.S., Vettori M.H.P.B., Monti R., Contiero J. (2014): Cyclodextrin production by Bacillus lehensis isolated from cassava starch: Characterisation of a novel enzyme. Czech J. Food Sci., 32: 48-53.
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 β-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°C and 8.0, respectively. The Michaelis-Menten constant was 8.62 g/l during maximum velocity of 0.858 g/l.h.Keywords:
cyclodextrin glycosyltransferase;affinity chromatography; purification