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Glycation improves the thermostability of trypsin and chymotrypsin
Author(s) -
Pham Van Thong,
Ewing Erin,
Kaplan Harvey,
Choma Christin,
Hefford Mary Alice
Publication year - 2008
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.21919
Subject(s) - thermostability , chemistry , trypsin , glycation , chymotrypsin , biochemistry , enzyme , denaturation (fissile materials) , chromatography , nuclear chemistry , receptor
A novel glycation procedure, in vacuo glycation, was used to attach glucose covalently to the lysine residues of trypsin and chymotrypsin. Glycated trypsin and glycated chymotrypsin have greatly increased thermostability compared to the native enzymes. For example, glycated bovine trypsin, incubated at 50°C and pH 8.0 for 3 h, retained more than 50% of its original activity whereas the native enzyme was inactivated under the same conditions. Similarly, after incubation at 50°C and pH 8.0, glycated bovine chymotrypsin retained 45% of its original activity and the native enzyme was inactivated. Glycated porcine trypsin is exceptionally thermostable and could be used to digest native ribonuclease at 70°C without the need for prior denaturation. The apparent increase in the thermal stability of the glycated proteins observed in activity measurements is also reflected by an increase in the T m values determined with differential scanning calorimetry (DSC) and circular dichroism (CD). The glycation does not alter the activity or specificity of these enzymes. Biotechnol. Bioeng. 2008;101: 452–459. © 2008 Wiley Periodicals, Inc.