Determinação das condições de separação e purificação de ß-glicosidase a partir de complexo enzimatico

AUTOR(ES)
DATA DE PUBLICAÇÃO

1996

RESUMO

Cellulose is a very common natural substance. The hydrolysis of this material is caused by an enzyme complex, acting synergistically, known as cellulase. In such a complex, the enzyme β-glucosidase (EC 3.2.1.21) is responsible for the final step of hydrolysis to form glucose. This enzyme has numerous industrial applications. Particularly important among applications in the food industry we may include the lightening of juices and the hydrolysis of vegetables for production of purees, for infants and elderly. This work was carried out to try to find the parameters involved in the separation and purification of , β-glucosidase, from an enzymatic complex called Cellulase Tr (Cellumax), and then, to apply them in the CARE (Continuous Affinity Recycle Extraction) system. The purification process began with gel filtration chromatography and followed by ion-exchange chromatography. The best results were obtained using CM-Sephadex C-50. The peak fractions obtained with ion-exchange cromatography were concentrated and applied to a Superose 12 R column in a FPLC system to confirm the purity of the , β-glucosidase. For the material obtained with Superose, a PAGE/SDS analysis was undertaken to garantee the purity of the enzime. The results obtained make possible the application of CARE process to the separation and purification of β-glucosidase. In addition, from the results it is possible to cal¬culate the kinetic constants involved in the resin adsorption-desorption used in the CARE system.

ASSUNTO(S)

beta-glicosidase - purificação celulose separação

ACESSO AO ARTIGO

Documentos Relacionados