title: Functional stabilization of trypsin by conjugation with beta-cyclodextrin-modified carboxymethylcellulose.
abstract: Bovine pancreatic trypsin was chemically modified by a beta-cyclodextrin-carboxymethylcellulose polymer using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide as coupling agent. The conjugate retained 110% and 95% of the initial esterolytic and proteolytic activity, respectively, and contained about 2 mol of polymer per mol of trypsin. The optimum temperature for trypsin was increased to 8 degrees C after conjugation. The thermostability of the enzyme was increased to about 16 degrees C after modification. The conjugate prepared was also more stable against thermal incubation at different temperatures ranging from 45 degrees C to 60 degrees C. In comparison with native trypsin, the polymer-enzyme complex was more resistant to autolytic degradation at pH 9.0, retaining about 65% of the initial activity after 3h incubation. In addition, modification protected trypsin against denaturation in the presence of sodium dodecylsulfate.
authors.affiliation: Enzyme Technology Group, Center for Biotechnological Studies, University of Matanzas, Matanzas, Cuba.
authors.forename: Maria L
authors.initials: ML
authors.lastname: Villalonga
authors.forename: Michael
authors.initials: M
authors.lastname: Fernández
authors.forename: Alex
authors.initials: A
authors.lastname: Fragoso
authors.forename: Roberto
authors.initials: R
authors.lastname: Cao
authors.forename: Reynaldo
authors.initials: R
authors.lastname: Villalonga
mesh_terms: D002266:Carboxymethylcellulose Sodium / Q000737:chemistry*; D003505:Cyclodextrins / Q000737:chemistry*; D004789:Enzyme Activation; D004795:Enzyme Stability; D007527:Isoenzymes / Q000138:chemical synthesis / Q000737:chemistry; D007700:Kinetics; D046911:Macromolecular Substances; D011485:Protein Binding; D011489:Protein Denaturation; D013696:Temperature; D014357:Trypsin / Q000138:chemical synthesis / Q000737:chemistry*; D047392:beta-Cyclodextrins
chemical_list: D003505:Cyclodextrins; D007527:Isoenzymes; D046911:Macromolecular Substances; D047392:beta-Cyclodextrins; D014357:Trypsin; C031215:betadex; D002266:Carboxymethylcellulose Sodium