Self-renaturing enzymes: Design of an enzyme-chaperone chimera as a new approach to enzyme stabilization

Lisa M. Bergeron, Lizabeth Gomez, Timothy A. Whitehead, Douglas S. Clark

Research output: Contribution to journalArticle

  • 24 Citations

Abstract

Molecular chaperones in aqueous-organic mixtures can broaden the utility of biocatalysis by stabilizing enzymes in denaturing conditions. We have designed a self-renaturing enzyme-chaperone chimera consisting of penicillin amidase and a thermophilic chaperonin that functions in aqueous-organic mixtures. The flexible linker separating the enzyme and chaperone domains was optimized and the design was extended to incorporate a chitin binding domain to facilitate immobilization of the chimera to a chitin support. The initial specific activity of penicillin amidase was not compromised by the enzymechaperone fusion or by immobilization. The total turnover number of immobilized chimera for amoxicillin synthesis in aqueous-methanol mixtures was 2.8 times higher after 95 h than the total turnover number of the immobilized penicillin amidase lacking a chaperone domain. Similarly, in 32% methanol the soluble chimera was active for over three times longer than the enzyme alone. This approach could easily be extended to other enzyme systems.

Original languageEnglish (US)
Pages (from-to)1316-1322
Number of pages7
JournalBiotechnology and Bioengineering
Volume102
Issue number5
DOIs
StatePublished - Apr 1 2009
Externally publishedYes

Profile

Enzymes
Penicillin Amidase
Chitin
Methanol
Agglutination Tests
Biocatalysis
Chaperonins
Molecular Chaperones
Amoxicillin
Fusion reactions
Stabilization
Cerebellar Ataxia

Keywords

  • Antibiotic synthesis
  • Chaperone
  • Fusion protein
  • Organic solvent
  • Protein folding

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Cite this

Self-renaturing enzymes : Design of an enzyme-chaperone chimera as a new approach to enzyme stabilization. / Bergeron, Lisa M.; Gomez, Lizabeth; Whitehead, Timothy A.; Clark, Douglas S.

In: Biotechnology and Bioengineering, Vol. 102, No. 5, 01.04.2009, p. 1316-1322.

Research output: Contribution to journalArticle

Bergeron, Lisa M.; Gomez, Lizabeth; Whitehead, Timothy A.; Clark, Douglas S. / Self-renaturing enzymes : Design of an enzyme-chaperone chimera as a new approach to enzyme stabilization.

In: Biotechnology and Bioengineering, Vol. 102, No. 5, 01.04.2009, p. 1316-1322.

Research output: Contribution to journalArticle

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