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Chemical glycosidation of trypsin with O ‐carboxymethyl‐poly‐β‐cyclodextrin: catalytic and stability properties
Author(s) -
Villalonga Maria L.,
Reyes Gretel,
Fragoso Alex,
Cao Roberto,
Fernández Leyden,
Villalonga Reynaldo
Publication year - 2005
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1042/ba20040115
Subject(s) - thermostability , chemistry , trypsin , carboxymethyl cellulose , autolysis (biology) , conjugate , thermal stability , cyclodextrin , enzyme , chemical modification , polymer chemistry , organic chemistry , sodium , mathematical analysis , mathematics
The polysaccharide O ‐carboxymethyl‐poly‐β‐cyclodextrin was synthesized (molecular mass 13000 Da, 40% carboxy groups) and attached to the surface of bovine pancreatic trypsin. The resulting neoglycoenzyme retained high proteolytic and esterolytic activity and contained approx. 1.0 mol of polymer/mol of enzyme. The optimum temperature for trypsin activity was increased by 10°C after this transformation. Thermostability of the polymer–enzyme complex was increased by about 14°C over 10 min incubation. The conjugate was also more resistant to thermal inactivation at different temperatures, ranging from 45 to 60°C, demonstrating the influence of supramolecular and polymer–protein electrostatic interactions on trypsin thermostabilization. Additionally, the conjugate was 36‐fold more resistant to the action of the anionic surfactant SDS. This modification also protected the enzyme from autolysis at alkaline pH.