Immobilization of penicillin acylase in porous beads of polyacrylamide gel
- 1 September 1991
- journal article
- Published by Springer Nature in Applied Biochemistry and Biotechnology
- Vol. 30 (3) , 265-272
- https://doi.org/10.1007/bf02922030
Abstract
A procedure is described for the immobilization of benzylpenicillin acylase fromEscherichia coli within uniformly spherical, porous polyacrylamide gel beads. Aqueous solutions of the enzyme and sodium alginate and of acrylamide monomer, N,N'-methylene-bis-acrylamide, N,N,N',N'-tetramethylethylenediamine (TEMED) and sodium alginate are cooled separately, mixed, and dropped immediately into ice-cold, buffered calcium formate solution, pH 8.5, to give calcium alginate-coated beads. The beads are left for 30-60 min in the cold calcium formate solution for polyacrylamide gel formation. The beads are then treated with a solution of glutaraldehyde and the calcium alginate subsequently leached out with a solution of potassium phosphate. Modification of the native enzyme with glutaraldehyde results in a slight enhancement in the rate of hydrolysis of benzylpenicillin at pH 7.8 and 0.05M substrate concentration. The enzyme entrapped in porous polyacrylamide gel beads shows no measurable diffusional limitation in stirred reactors, catalyzing the hydrolysis of the substrate at a rate comparable to that of the glutaraldehyde-modified native enzyme. The immobilized enzyme preparation has been used in batch mode over 90 cycles without any apparent loss in hydrolytic activity.Keywords
This publication has 5 references indexed in Scilit:
- Immobilization of Saccharomyces uvarum cells in porous beads of polyacrylamide gel for ethanolic fermentationApplied Microbiology and Biotechnology, 1988
- Kinetic behavior of immobilized Penicillin acylaseBiotechnology & Bioengineering, 1980
- The isolation and kinetics of penicillin amidase from Escherichia coliBiochimica et Biophysica Acta (BBA) - Enzymology, 1972
- Automated Colorimetric Determination of 6-Aminopenicillanic Acid in Fermentation Media.Analytical Chemistry, 1965
- PROTEIN MEASUREMENT WITH THE FOLIN PHENOL REAGENTJournal of Biological Chemistry, 1951