Oral Presentation 7-02

 

Phenylboronate-Chitosan Resins for Adsorption of

Beta-amylase from Soybean Extracts

 

E.J. Arruda1 and C.C. Santana2

 

1Pharmacy and Biochemistry

Catholic University Dom Bosco, UCBD

Campo Grande, MS Brazil

C.P 100, CEP 79002-250

 

2School of Chemical Engineering

DPB, UNICAMP

Campinas, SP  Brazil

C.P. 6066,  CEP 13083-970

 

Telephone:  (55) 19-37883917; Fax:  (55) 19-37883918; Email:  santana@feq.unicamp.br

 

Isolation and purification of bioproducts starting from crude extracts can be obtained by methods based on affinity for a certain molecule that binds specifically and reversibly to a ligand molecule immobilized in a porous matrix.  The b-amylase  (b-1,4-glucan maltohydrolase, E.C 3.2.1.2) is an exo-enzyme that removes units of maltose of non-reduced terminals of the polysaccharides chains.  There is great industrial interest for the enzyme in the production of rich syrups in maltose, and interest for medical use as an intensifier of the fagocitose in carcinomas.  Presented here are the kinetic curves and the isotherms of adsorption of the crude extracts, as well as the breakthrough curves for a frontal chromatographic separation method of a commercial sample of beta-amylase from soybean in the interaction with the adsorbents prepared starting from a chitosan matrix with 3-aminophenylboronic acid.  Those results were compared to similar data obtained with an agarose based commercial gel (Matrex gel PBA 30 - Amicon, USA).  The adsorbent’s behavior was markedly different from that of phenylboronate-chitosan (PBC), to which the enzyme was rapidly adsorbed and which required much more drastic elution conditions.

 

Keywords: purification; beta-amylase;.soybean; phenyl-boronate; chitosan.

 

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