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PublicationPROCESS BIOCHEMISTRY
Year2013
Volume48
Pages1054-1058
International

Optimization of the Immobilization of Sweet Potato Amylase Using Glutaraldehyde-Agarose Support. Characterization of the immobilized enzyme.

Authors:Roberto Fernandez-Lafuente
Groups of research:Optimización de biocatalizadores y bioprocesos enzimáticos
Olga Luisa Tavano*,a, Roberto Fernandez-Lafuenteb, Antonio José Goulartc, Rubens Montid
 
a Department of Nutrition, ICS – Universidade Federal do Triângulo Mineiro. Rua Getulio Guaritá, 159 – Uberaba – MG, Brazil, CEP 38025 360, e-mail address: tavanool@yahoo.com.br
b Department of Biocatalysis, ICP - CSIC. Campus UAM-CSIC. Cantoblanco, ZC 28049, Madrid, Spain, e-mail address: rfl@icp.csic.es
c Department of Food Nutrition, Faculty of Pharmaceutical Sciences, UNESP – São Paulo State University. Rodovia Araraquara-Jaú, Km1, 14840-000, Araraquara, SP, Brazil, e-mail address: antoniogoulart@yahoo.com
d Department of Food Nutrition, Faculty of Pharmaceutical Sciences, UNESP – São Paulo State University. Rodovia Araraquara-Jaú, Km1, 14840-000, Araraquara, SP, Brazil, e-mail address: montiru@fcfar.unesp.br

A simplified procedure for the preparation of immobilized beta-amylase using non-purified extract from fresh sweet potato tubers is established in this paper, using differently activated agarose supports. Beta-amylase glutaraldehyde derivative was the preparation with best features, presenting improved temperature and pH stability and activity. The possibility of reusing the amylase was also shown, when this immobilized enzyme was fully active for five cycles of use. However, immobilization decreased enzyme activity to around 15%. This seems to be mainly due to diffusion limitations of the starch inside the pores of the biocatalyst particles. A fifteen-fold increase in the Km was noticed, while the decrease of Vmax was only 30% (10.1 U/mg protein and 7.03 U/mg protein for free and immobilized preparations, respectively).
Keywords:Beta-amylase; Immobilization; Sweet potato; Agarose; Glutaraldehyde
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