Ion exchange immobilization of changed β-galactosidase fusions for lactose hydrolysis

The use of charged peptides fused to enzymes for immobilization onto ion‐exchange membranes was explored for the enzyme ×‐galactosidase. The additional charged peptides, containing 1, 5, 11, and 16 aspartates, fused to ×‐galactosidase, for the most part did not interfere with the kinetic behavior fo...

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Bibliographic Details
Published in:Biotechnology and bioengineering 1994-09, Vol.44 (6), p.745-752
Main Authors: Heng, Meng H., Glatz, Charles E.
Format: Article
Language:English
Subjects:
charged fusions
ion exchange
whey hydrolysis
β-galactosidase immobilization
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Summary:The use of charged peptides fused to enzymes for immobilization onto ion‐exchange membranes was explored for the enzyme ×‐galactosidase. The additional charged peptides, containing 1, 5, 11, and 16 aspartates, fused to ×‐galactosidase, for the most part did not interfere with the kinetic behavior for lactose hydrolysis. There was a 2‐fold decline in Vm for the 16‐aspartate fusion, but the others were quite similar to the wild type enzyme (BGWT). BGWT and the fusions all retained approximately 50% of their activities when adsorbed onto ion‐exchange membranes. In contrast to BGWT, the enhanced binding strength of the 11 aspartate fusion provided the ability to hydrolyze whey permeate at 0.3 M ionic strength without enzyme leakage, and to immobilize the enzyme directly from diluted cell extract with 83% purity. © 1994 John Wiley & Sons, Inc.
ISSN:0006-3592
1097-0290
DOI:10.1002/bit.260440611