Abstract

Background: Chemistry and particularly enzymology at surfaces is a topic of rapidly growing interest, both in terms of its role in biological systems and its application in biocatalysis. Existing protein immobilization approaches, including noncovalent or covalent attachments to solid supports, have difficulties in controlling protein orientation, reducing nonspecific absorption and preventing protein denaturation. New strategies for enzyme immobilization are needed that allow the precise control over orientation and position and thereby provide optimized activity. Methodology/Principal Findings: A method is presented for utilizing peptide ligands to immobilize enzymes on surfaces with improved enzyme activity and stability. The appropriate peptide ligands have been rapidly selected from high-density arrays and when desirable, the peptide sequences were further optimized by single-point variant screening to enhance both the affinity and activity of the bound enzyme. For proof of concept, the peptides that bound to β-galactosidase and optimized its activity were covalently attached to surfaces for the purpose of capturing target enzymes. Compared to conventional methods, enzymes immobilized on peptide-modified surfaces exhibited higher specific activity and stability, as well as controlled protein orientation. Conclusions/Significance: A simple method for immobilizing enzymes through specific interactions with peptides anchored on surfaces has been developed. This approach will be applicable to the immobilization of a wide variety of enzymes on surfaces with optimized orientation, location and performance, and provides a potential mechanism for the patterned self-assembly of multiple enzymes on surfaces.

Original languageEnglish (US)
Article numbere18692
JournalPLoS One
Volume6
Issue number4
DOIs
StatePublished - 2011

Fingerprint

Enzyme immobilization
immobilized enzymes
Immobilization
peptides
Peptides
Enzymes
enzymes
Proteins
Biocatalysis
Galactosidases
enzymology
Ligands
enzyme stability
galactosidases
Protein Denaturation
Enzyme Stability
Immobilized Enzymes
Denaturation
proteins
Enzyme activity

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Peptide-modified surfaces for enzyme immobilization. / Fu, Jinglin; Reinhold, Jeremy; Woodbury, Neal.

In: PLoS One, Vol. 6, No. 4, e18692, 2011.

Research output: Contribution to journalArticle

Fu, Jinglin ; Reinhold, Jeremy ; Woodbury, Neal. / Peptide-modified surfaces for enzyme immobilization. In: PLoS One. 2011 ; Vol. 6, No. 4.
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