Transglutaminase-catalyzed synthesis of trypsin-cyclodextrin conjugates: kinetics and stability properties.
Bovine pancreatic trypsin was modified by the mono-6-amino-6-deoxy derivatives of alpha-, beta-, and gamma-cyclodextrin through a transglutaminase-catalyzed reaction. The trypsin-cyclodextrin conjugates, containing about 3 mol of oligosaccharide per mole of protein, were tested for their catalytic and stability properties. The specific esterolytic activity and the kinetics constants of trypsin were significantly improved following the transglutaminase-induced structural modifications. Trypsin-cyclodextrin conjugates were also found markedly (sixfold) more resistant to autolytic degradation at alkaline pH, and their thermal stability profile was improved by about 16 degrees C. Moreover, they were particularly resistant to heat inactivation when treated at different temperatures ranging from 45 degrees C to 70 degrees C for different periods of time.
Villalonga
Reynaldo
R
Enzyme Technology Group, Center for Biotechnological Studies, University of Matanzas, Autopista a Varadero, Matanzas, Cuba.
Fernández
Michael
M
Fragoso
Alex
A
Cao
Roberto
R
Di Pierro
Prospero
P
Mariniello
Loredana
L
Porta
Raffaele
R
D000818:Animals; D001120:Arginine / Q000031:analogs & derivatives* / Q000737:chemistry*; D002384:Catalysis; D002417:Cattle; D003067:Coenzymes / Q000138:chemical synthesis; D004789:Enzyme Activation; D004795:Enzyme Stability; D005964:Glucosyltransferases / Q000138:chemical synthesis* / Q000737:chemistry; D006358:Hot Temperature; D007700:Kinetics; D046911:Macromolecular Substances; D009097:Multienzyme Complexes / Q000138:chemical synthesis* / Q000737:chemistry; D011522:Protons; D013379:Substrate Specificity; D011503:Transglutaminases / Q000737:chemistry*; D014357:Trypsin / Q000138:chemical synthesis* / Q000737:chemistry
D003067:Coenzymes; D046911:Macromolecular Substances; D009097:Multienzyme Complexes; D011522:Protons; D001120:Arginine; C009727:benzoylarginine ethyl ester; D011503:Transglutaminases; D005964:Glucosyltransferases; C019963:cyclomaltodextrin glucanotransferase; D014357:Trypsin