Optimization of the immobilization of sweet potato amylase using glutaraldehyde-agarose support. Characterization of the immobilized enzyme
| Autor: | Roberto Fernandez-Lafuente, Olga Luisa Tavano, Antonio José Goulart, Rubens Monti |
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| Přispěvatelé: | Conselho Nacional de Desenvolvimento Científico e Tecnológico (Brasil), Ministerio de Ciencia e Innovación (España), Universidade Federal do Triângulo Mineiro (UFTM), ICP-CSIC, Universidade Estadual Paulista (Unesp) |
| Rok vydání: | 2013 |
| Předmět: |
Immobilized enzyme
Starch Bioengineering Applied Microbiology and Biotechnology Biochemistry Industrial and Manufacturing Engineering Glutaraldehyde Glutaraldehyde-agarose chemistry.chemical_compound Immobilization Glutaraldehydes Radioactive waste vitrification Agarose Enzyme immobilization Amylase Ipomoea batatas Beta-amylase Engineering (miscellaneous) Solanum tuberosum Biocatalyst particle Chromatography biology Chemistry Organic Chemistry Proteins Ph stability Enzyme assay Simplified procedure Diffusion limitations Biocatalysis Sweet potato Amylases biology.protein |
| Zdroj: | Digital.CSIC. Repositorio Institucional del CSIC instname Scopus Repositório Institucional da UNESP Universidade Estadual Paulista (UNESP) instacron:UNESP |
| Popis: | 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−1 protein and 7.03 U mg−1 protein for free and immobilized preparations, respectively). We would like to thank CNPq and PADC-FCFAr for financial support. We gratefully recognize the support from the Spanish Government, grant CTQ2009-07568. |
| Databáze: | OpenAIRE |
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