Orientation specific immobilization of organophosphorus hydrolase on magnetic particles through gene fusion

J. Wang, D. Bhattacharyya, Leonidas G Bachas

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

Recombinant DNA technology has been utilized to fuse an octapeptide, Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys (FLAG), to the C-terminus of organophosphorus hydrolase (OPH, EC 3.1.8.1), an enzyme capable of hydrolyzing organophosphate compounds, such as insecticides and nerve gas agents. The recombinant OPH-FLAG was immobilized onto magnetic beads coated with protein A in the following ways: (a) site-directly through a monoclonal antibody (MAb) specific for the FLAG peptide; (b) through the MAb that was randomly tethered to the beads using glutaraldehyde; (c) randomly by cross-linking OPH-FLAG to protein-coated beads using glutaraldehyde. Kinetic studies demonstrated that the site-directly immobilized enzyme maintained the highest catalytic efficiency. The orientation specific immobilization strategy described in this article can be applied to other proteins, and therefore, it may find potential applications in the design of biosensors, biocatalysts, and bioreactors having immobilized proteins as their biorecognition elements.

Original languageEnglish (US)
Pages (from-to)700-705
Number of pages6
JournalBiomacromolecules
Volume2
Issue number3
DOIs
StatePublished - 2001
Externally publishedYes

Fingerprint

Aryldialkylphosphatase
Hydrolases
Gene Fusion
Glutaral
Immobilization
Fusion reactions
Genes
Monoclonal Antibodies
Immobilized Proteins
Proteins
Genetic engineering
Immobilized Enzymes
Monoclonal antibodies
Recombinant DNA
Organophosphates
Staphylococcal Protein A
Biosensing Techniques
Bioreactors
Electric fuses
Enzymes

ASJC Scopus subject areas

  • Organic Chemistry
  • Biochemistry, Genetics and Molecular Biology(all)
  • Polymers and Plastics
  • Materials Chemistry

Cite this

Orientation specific immobilization of organophosphorus hydrolase on magnetic particles through gene fusion. / Wang, J.; Bhattacharyya, D.; Bachas, Leonidas G.

In: Biomacromolecules, Vol. 2, No. 3, 2001, p. 700-705.

Research output: Contribution to journalArticle

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