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Characterization of cyclodextrin glycosyltransferase from Bacillus firmus strain No. 37

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Abstract

The enzyme cyclod extringly cosyltransferase (CGTase), EC2.4.1.19, which produces cyclodextrins (CDs) from starch, was obtained from Bacillus firmus strain no. 37 isolated from Brazilian soil and characterized in the soluble form using as substrate 100 g/L of maltodextrin in 0.05 M Tris-HCl buffer, 5 mM CaCl2, and appropriate buffers. Enzymatic activity and its activation energy were determined as a function of temperature and pH. The activation energy for the production of β- and γ-CD was 7.5 and 9.9 kcal/mol, respectively. The energy of deactivation was 39 kcal/mol. The enzyme showed little thermal deactivation in the temperature range of 35–60°C, and Arrhenius-type equations were obtained for calculating the activity, deactivation, and half-life as a function of temperature. The molecular weight of the enzyme was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis, giving 77.6k Da. Results for CGTase activity as a function of temperature gave maximal activity for the production of β-CD at 65°C, pH 6.0, and 7 1.5 mmol of β-CD/(min·mg of protein), whereas for γ-CD it was 9.1 m mol of γ-CD/(min·mg of protein) at 70°C and pH 8.0. For long contact times, the bestuse of the enzymatic activity occurs at 60°C oratalower temperature, and the reaction pH may be selected to increase the vield of a desired CD.

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Authors and Affiliations

  1. Pharmacy and Pharmacology Department, State University of Maringá, Av. Colombo, 5790, BL D-90, 87020-900, Maringá, PR, Brazil

    Graciette Matioli

  2. Chemical Engineering Department, State University of Maringá, Av. Colombo, 5790, BL D-90, 87020-900, Maringá, PR, Brazil

    Gisella M. Zanin & Flávio F. De Moraes

Authors
  1. Graciette Matioli
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  2. Gisella M. Zanin
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  3. Flávio F. De Moraes
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Correspondence to Flávio F. De Moraes.

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Matioli, G., Zanin, G.M. & De Moraes, F.F. Characterization of cyclodextrin glycosyltransferase from Bacillus firmus strain No. 37. Appl Biochem Biotechnol 91, 643–654 (2001). https://doi.org/10.1385/ABAB:91-93:1-9:643

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