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Ion exchange immobilization of changed β‐galactosidase fusions for lactose hydrolysis
Author(s) -
Heng Meng H.,
Glatz Charles E.
Publication year - 1994
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260440611
Subject(s) - chemistry , hydrolysis , membrane , lactose , ionic strength , enzyme , ion exchange , chromatography , adsorption , enzymatic hydrolysis , biochemistry , ion , organic chemistry , aqueous solution
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 V m 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.