ISSN:
1433-4909
Keywords:
Key wordsβ-Hydroxyectoine
;
Betaine
;
RNase A
;
Differential scanning calorimetry
;
Enzyme stabilization
;
Osmolytes
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Abstract Thermodynamic aspects of protein stabilization by two widespread naturally occurring osmolytes, β-hydroxyectoine and betaine, were studied using differential scanning calorimetry (DSC) and bovine ribonuclease A (RNase A) as a model protein. The osmolyte β-hydroxyectoine purified from Marinococcus was found to be a very efficient stabilizer. At a concentration of 3 M it increased the melting temperature of RNase A (T m ) by more than 12 K and gave rise to a stability increase of 10.6 kJ/mol at room temperature. The heat capacity difference between the folded and unfolded state (ΔC p ) was found to be significantly increased. Betaine stabilized RNase A only at concentrations less than 3 M. Also, here ΔC p was found to be increased. Calculation of the number of water molecules that additionally bind to unfolded RNase A resulted in surprisingly low numbers for both osmolytes. The significant stabilization of RNase A by β-hydroxyectoine makes this osmolyte an interesting stabilizer in biotechnological processes in which enzymes are applied in the presence of denaturants or at high temperature.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/s007920050116
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