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Transglutaminase‐catalyzed synthesis of trypsin–cyclodextrin conjugates: Kinetics and stability properties
Author(s) -
Villalonga Reynaldo,
Fernández Michael,
Fragoso Alex,
Cao Roberto,
Di Pierro Prospero,
Mariniello Loredana,
Porta Raffaele
Publication year - 2003
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.10520
Subject(s) - trypsin , chemistry , tissue transglutaminase , kinetics , cyclodextrin , conjugate , thermal stability , enzyme , oligosaccharide , catalysis , chromatography , biochemistry , organic chemistry , mathematical analysis , physics , mathematics , quantum mechanics
Bovine pancreatic trypsin was modified by the mono‐6‐amino‐6‐deoxy derivatives of α‐, β‐, and γ‐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°C. Moreover, they were particularly resistant to heat inactivation when treated at different temperatures ranging from 45°C to 70°C for different periods of time. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 81: 732–737, 2003.