CHISA 2008 - 18th International Congress of Chemical and Process Engineering (CD-ROM of Full Texts)info:eu-repo/semantics/publishedVersio

Gonçalves, M. P.

Rocha, Cristina M. R.

Teixeira, J. A.

Universidade do Minho: RepositoriUM

A comparative analysis on the efficiency of different carriers for trypsin immobilization   C. Rocha1,2, M. P. Gonçalves3, J. A. Teixeira11 IBB-Institute for Biotechnology and Bioengineering, Centre for Biological Engineering, Universidade do Minho, Campus de Gualtar 4710-057, Braga, Portugal; tel. +351 253 604400, e-mail: jateixeira@deb.uminho.pt; 2 Escola Superior de Tecnologia e Gestão, Instituto Politécnico de Viana do Castelo, Apartado 574, 4900-348 Viana do Castelo, Portugal; e-mail: cristina@estg.ipvc.pt; 3 REQUIMTE, Departamento de Engenharia Química, Faculdade de Engenharia, Universidade do Porto, Rua Dr. Roberto Frias, 4200-465 Porto, Portugal; e-mail: pilarg@fe.up.pt    Trypsin is a widely used enzyme for protein hydrolysis and can be used to improve functional and nutritional properties of foods. Its immobilization on solid carriers can offer several advantages over the free enzyme including easy handling, recovery from the reaction medium, reuse and operation in continuous reactors.  This work compares different carriers, namely zeolyte NaY, spent grains and magnetic polysiloxane-polyvinyl alcohol composite (POS-PVA), as potential candidates for trypsin immobilization. Covalent attachment to the carriers was tested using glutaraldehyde or glycydol. The efficiency of immobilization and activity, operation and storage stability of free and immobilized enzyme on the supports were evaluated.  Table 1: Immobilized protein and activity retention achieved with the different carriers Carrier Type of enzyme Immobilized protein (%) Activity (U/g carrier) Retention of activity (%) Zeolite Crude 46.8±0.9 35.6±1.5 66.8 Zeolite Purified 70.0±4.1 52.6±1.9 63.5 POS-PVA Crude 30.3±1.3 3.27±0.27 13.7 POS-PVA Purified 41.6±7.7 37.6±5.6 73.7 Glyoxyl spent grain Crude 68.7±1.0 8.25±0.96 10.5 Glyoxyl spent grain Purified 69.8±3.9 44.3±2.2 46.0  When a crude enzyme as used, the amount of immobilized protein achieved was high (up to 70 %) but the activity retention was low for all carriers except for trypsin crosslinked on zeolites, where it was satisfactory (Table 1). However, when a more purified enzyme from bovine pancreas was used with glyoxyl-spent grain or POS-PVA with glutaraldehyde, the activity retention was of 46 % and 73 % against 11 % and 9 % with crude enzyme.  The operational stability obtained was always close to 100 % while the storage stability of these immobilized enzymes was always above 80 %. Thus it can be stated that trypsin was successfully immobilized on spent grains by multipoint covalent attachment using glycidol and on POS-PVA functionalized with glutaraldehyde. The immobilized trypsin with the highest activity was however achieved with covalent binding through glutaraldehyde to silanized zeolite followed by crosslinking with glutaraldehyde, probably due to a positive effect of the zeolite on the enzyme activity.  

A comparative analysis on the efficiency of different carriers for trypsin immobilization

http://repositorium.sdum.uminho.pt/bitstream/1822/48389/1/document_46884_1.pdf

A comparative analysis on the efficiency of different carriers for trypsin immobilization

Abstract

Similar works

Full text

Available Versions

Universidade do Minho: RepositoriUM