title: Peptide-modified surfaces for enzyme immobilization.
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.
  
  
  RESULTS
  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
  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.
authors.affiliation: Center for Single Molecule Biophysics, Arizona State University, Tempe, Arizona, United States of America.
authors.forename: Jinglin
authors.initials: J
authors.lastname: Fu
authors.forename: Jeremy
authors.initials: J
authors.lastname: Reinhold
authors.forename: Neal W
authors.initials: NW
authors.lastname: Woodbury
mesh_terms: D000595:Amino Acid Sequence; D001671:Biochemistry / Q000379:methods*; D004800:Enzymes, Immobilized / Q000378:metabolism*; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D013499:Surface Properties; D001616:beta-Galactosidase / Q000378:metabolism*
chemical_list: D004800:Enzymes, Immobilized; D010455:Peptides; D001616:beta-Galactosidase