PBC Resins for Adsorption of β-Amylase 829
Applied Biochemistry and Biotechnology Vol. 105–108, 2003
Copyright © 2003 by Humana Press Inc.
All rights of any nature whatsoever reserved.
0273-2289/03/105-108/0829/$20.00
829
*Author to whom all correspondence and reprint requests should be addressed.
Phenylboronate-Chitosan Resins
for Adsorption of β-Amylase
from Soybean Extracts
EDUARDO J. ARRUDA
1
AND CESAR C. SANTANA
*,2
1
Pharmacy and Biochemistry, Dom Bosco Catholic University,
UCDB, C.P. 100, CEP 79117-900, Campo Grande, MS, Brazil; and
2
School of Chemical Engineering, DPB, UNICAMP, C.P. 6066,
CEP 13083-970, Campinas, SP, Brazil; E-mail: santana@feq.unicainp.br
Abstract
Isolation and purification of bioproducts from crude extracts can be
obtained by affinity methods based on reversible binding of a specific mol-
ecule to ligand immobilized in a porous matrix. In the present work,
nicrospheres based on chitosan matrix, which incorporated aminophenyl-
boronic acid as a derivative, were prepared and characterized, aimed at
developing a β-amylase adsorption process. Kinetic curves and adsorption
isotheriru of the crude extracts as well as the breakthrough curves for
a frontal chromatographic separation method of a commercial sample of
β-amylase from soybean are presented. These results were compared to simi-
lar data obtained with a comercial microspheres gel based-on agarose.
Index Entries: Purification; β-amylase; soybean; phenylboronate; chitosan.
Introduction
Interest in bioproducts has been increasing as a result of biotechno-
logical development. Industrial demands for new applications, specific-
ity, and renewability of products have also increased dramatically. Because
of the expansion of downstream processing in biotechnology, the methods
aimed at concentration and purification of enzymes and biopolymers after
their production from a variety of sources require new advances (1–3).
β-Amylase (β-1,4-glucanmaltohydrolase, EC 3.2.1.2) is an exoenzyrne that
removes units of nonreduced maltose terminals from polysaccharide
chains, producing β-maltose and β-limit dextrins. There is a considerable
industrial interest in this enzyme for the production of syrups rich in mal-