Cellulosic exopolysaccharide produced by Zoogloea sp as a film support for trypsin immobilisation

Trypsin (E.C. 3.4.21.4) was covalently immobilised onto a membrane of a cellulosic exopolysaccharide produced by Zoogloea sp. in sugarcane molasses. Carbonyl groups were introduced into the matrix by sodium metaperiodate oxidation and the enzyme was immobilised either directly or through bovine serum albumin (BSA) as a spacer. The trypsin-membrane and trypsin-BSA-membrane retained, respectively, 37.2% and 9.16% of the specific activity of the native enzyme acting on N-benzoil-DL-arginine-p-nitroanilide (BAPNA). No activity decrease was observed in both preparations after seven reutilisations as well as they showed to be more thermal stable than the native enzyme. The trypsin-BSA-i-nembrane presented the same initial activity (99%) after 54 days stored in 0.1 M Tris-HCl buffer, pH 8.0, at 4 degrees C but the trypsin-membrane lost 15% of activity. Furthermore, the trypsin-BSA-membrane lost 31% of activity after reuse at 9 days interval during 54 days of storage at 4 degrees C whereas the trypsin-membrane lost 69% of activity under the same conditions. These results showed an additional application for this biofilm, namely, to act as a reusable matrix for trypsin immobilisation and the presence of BSA improved the derivative performance. (c) 2006 Elsevier B.V. All rights reserved.