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CHEMICAL PROTEIN GLYCOSYLATION: A METHOD TO PREVENT PROTEIN INSTABILITIES ON BIOSENSOR APPLICATION
Author(s) -
Pagan Miraida,
Griebenow Kai
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.697.2
Subject(s) - glycosylation , subtilisin , chemistry , enzyme , biosensor , chemical modification , glycan , glycoprotein , n linked glycosylation , covalent bond , chemical stability , biochemistry , biophysics , organic chemistry , biology
The development of small and robust sensors with low electrical power requirement will be essential to monitor astronaut physical fitness and condition in order to prevent and minimize adverse health effects that could result from space flight. Even though there are many advances reported recently using enzymes (e.g., alcohol dehydrogenase, glucose oxidase) as the principal sensing element to monitor specific biochemical analytes, an efficient methodology to prevent short lifetime and poor loading of the protein onto the sensor is lacking. These limitations are related to enzyme stability problems. Herein, we seek to improve the understanding of enzyme stabilization in order to increase the lifetime and performance of enzyme biosensors by employing the glycosylation technique. Covalent chemical glycosylation consists in decorating the enzyme with glycans which will increase protein thermodynamic stability and prevent unfolding by caging effects. Furthermore, attachment of the modified protein to a nano‐material will increase the enzyme loading and improved reaction kinetics In order to study the effect of chemical glycosylation it was used a simple enzyme model, Subtilisin. The central hypothesis of the research is that covalent chemical modification of the enzyme with glycans will reduce conformational motions and increase its thermodynamic stability thus positively affecting long‐term stability