Czech J. Food Sci., 2014, 32(1):48-53 | DOI: 10.17221/432/2012-CJFS

Cyclodextrin production by Bacillus lehensis isolated from cassava starch: Characterisation of a novel enzymeOriginal Paper

Kate Cristina Blanco1, Flávio Faria De Moraes2, Natalia Sozza Bernardi1, Mary Helen Palmuti Braga Vettori1, Rubens Monti3, Jonas Contiero1
1 Department of Biochemistry and Microbiology, Biological Sciences Institute, UNESP - University Estadual Paulista, Rio Claro, Brazil
2 Chemical Engineering Department, UEM - University Estadual de Maringá, Maringá, Brazil
3 Department of Food and Nutrition, Faculty of Pharmaceutical Sciences, UNESP - University Estadual Paulista, Araraquara, Brazil

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

Published: February 28, 2014  Show citation

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Blanco KC, De Moraes FF, Bernardi NS, Vettori MHPB, Monti R, Contiero J. Cyclodextrin production by Bacillus lehensis isolated from cassava starch: Characterisation of a novel enzyme. Czech J. Food Sci. 2014;32(1):48-53. doi: 10.17221/432/2012-CJFS.
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