Stabilization and re-activation of trapped enzyme by immobilized heat shock protein and molecular chaperones

  • Yang Y
  • Zeng J
  • Gao C
 et al. 
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The potential of using immobilized Heat Shock Protein 70 (HSP 70) in combination with other molecular chaperones to ameliorate problems of enzyme denaturation was investigated. Firefly luciferase was used as a model enzyme due to its sensitivity to thermal denaturation, and the availability of a sensitive chemiluminescent assay method for determination of relative activity of this enzyme. Control experiments and development of effective combinations of HSP with other chaperones involved re-activation of enzyme in bulk solution. A combination of HSP 70, α-crystallin and reticulocyte lysate (RL) in bulk solution were found to re-activate soluble firefly luciferase to about 60% of the initial activity after the enzyme activity had been reduced to less than 2% by thermal denaturation. HSP 70 that was covalently immobilized onto glass surfaces was also able to re-activate denatured enzyme that was in bulk solution. Over 30% of the initial activity could be regained from heat denatured enzyme when using immobilized HSP in the presence of other chaperones. The activity of soluble enzyme decayed to negligible values in a period of days when stored at room temperature. In the presence of immobilized HSP and chaperones, activity stabilized at about 10% of the initial activity even after many weeks. The results suggest that immobilized molecular chaperones such as HSP 70 may provide some potential for stabilization and re-activation of enzymes that are trapped in thin aqueous films for applications in biosensors and reactors. © 2002 Elsevier Science B.V. All rights reserved.

Author-supplied keywords

  • Firefly luciferase
  • HSP 70
  • Heat shock protein
  • Immobilization
  • Molecular chaperones
  • Re-activation
  • Reticulocyte lysate
  • α-Crystallin

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  • Yunhui Yang

  • Jiang Zeng

  • Chunguang Gao

  • Ulrich J. Krull

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