Advertisement

Biotechnology Techniques

, Volume 13, Issue 9, pp 637–641 | Cite as

Lysozyme reactivation using immobilized molecular chaperonin GroEL

  • Xiao-Yan Dong
  • Hui Yang
  • Yan Sun
Article

Abstract

The molecular chaperonin, GroEL, was immobilized to a porous matrix and used to reactivate denatured lysozyme. The maximum reactivation yield was obtained at 37 °C and pH 6–8 and about 90% activity of the denatured lysozyme was restored under the conditions. The coupling density of GroEL had little effect on the chaperoning activity of GroEL up to 48 mg g−1 gel. The immobilized GroEL was reusable, indicating the possibility of using it on a large scale for the refolding of proteins.

GroEL immobilization lysozyme reactivation refolding 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Altamirano MM, Golbik R, Zahn R, Buckle AM, Fersht AR (1997) Refolding chromatography with immobilized mini-chaperones. Proc. Natl. Acad. Sci. USA 94: 3576-3578.Google Scholar
  2. Fisher MT (1992) Promotion of the in vitro renaturation of dodecametric glutamine synthetase from Escherichia coli in the presence of GroEL (chaperonin-60) and ATP. Biochemistry 31: 3955-3963.Google Scholar
  3. Guise AD, Chaudhuri JB (1998) Recovery and reuse of the molecular chaperone GroEL for in vitro protein refolding. Biotechnol. Prog. 14: 343-346.Google Scholar
  4. Hartl F-U (1996) Molecular chaperones in cellular protein folding. Nature 381: 571-580.Google Scholar
  5. Imoto T, Yagishita K (1971) A simple activity measurement of lysozyme. Agric. Biol. Chem. 35: 1154-1156.Google Scholar
  6. Ishii Y, Teshima T, Kondo A, Murakami K, Sonezaki S, Ogawa H, Kato Y, Fukuda H (1997) Operation conditions of enzyme refolding by chaperonin and recycle system using ultrafiltration. Chem. Eng. J. 65: 151-157.Google Scholar
  7. Jenkins AJ, March JB, Oliver IR, Masters M (1986) DNA fragment containing the groE genes can suppress mutations in Escherichia coli dnaA genes. Mol. Gen. Genet. 202: 446-454.Google Scholar
  8. Mendoza JA, Rogers E, Lorimer GH, Horowitz PM (1991) Chaperonins facilitate the in vitro folding of monometric mitochondrial rhodanase. J. Biol. Chem. 266: 13044-13049.Google Scholar
  9. Sadana A (1995) Protein refolding and inactivation during bioseparation: Bopprocessing implications. Biotechnol. Bioeng. 48: 481-489.Google Scholar
  10. Teshima T, Kondo A, Fukuda H (1997) Reactivation of thermally inactivated enzymes by free and immobilized chaperonin GroEL/ES. Appl. Microbiol. Biotechnol. 48: 41-46.Google Scholar

Copyright information

© Kluwer Academic Publishers 1999

Authors and Affiliations

  • Xiao-Yan Dong
    • 1
  • Hui Yang
    • 1
  • Yan Sun
    • 1
  1. 1.Department of Biochemical EngineeringTianjin UniversityTianjinPeople's Republic of China

Personalised recommendations