Selective Functionalization of Microgels with Enzymes by Sortagging

Bioconjug Chem. 2019 Nov 20;30(11):2859-2869. doi: 10.1021/acs.bioconjchem.9b00568. Epub 2019 Oct 11.

Abstract

Enzyme immobilization has been widely used to improve the stability and recyclability of enzymes in industrial processes. In this work, a sortase-mediated and therefore selective covalent immobilization strategy (sortagging) for enzymes on microgels (GelZyms) was investigated. Aqueous microgels were synthesized from poly(N-vinylcaprolactam)/glycidyl methacrylate (PVCL/GMA) and tagged with the sortase A recognition peptide sequence (LPETG) or its nucleophilic counterpart-tag (GGG). General applicability and selective immobilization were confirmed by subsequent sortagging of five different enzymes (Bacillus subtilis lipase A (BSLA), Yersinia mollaretii phytase (Ym-phytase), Escherichia coli copper efflux oxidase (CueO laccase), cellulase A2, and Bacillus megaterium monooxygenase P450 BM3). The latter was performed directly from the cell lysate to ensure cost-effective immobilization. All five immobilized enzymes were catalytically active and could be recycled (e.g., laccase CueO and monooxygenase P450 BM3 F87A; >55% residual activity after six cycles). Application potential was demonstrated by using CueO decorated microgels for bleaching of the synthetic dye indigo carmine.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 6-Phytase / chemistry
  • 6-Phytase / metabolism
  • Aminoacyltransferases / chemistry
  • Aminoacyltransferases / metabolism
  • Bacteria / growth & development
  • Bacteria / metabolism*
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / metabolism*
  • Cellulase / chemistry
  • Cellulase / metabolism
  • Cysteine Endopeptidases / chemistry
  • Cysteine Endopeptidases / metabolism
  • Enzymes, Immobilized / chemistry
  • Enzymes, Immobilized / metabolism*
  • Laccase / chemistry
  • Laccase / metabolism
  • Lipase / chemistry
  • Lipase / metabolism
  • Microgels / chemistry*
  • Oxidoreductases / chemistry
  • Oxidoreductases / metabolism

Substances

  • Bacterial Proteins
  • Enzymes, Immobilized
  • Microgels
  • Oxidoreductases
  • Laccase
  • copper oxidase
  • Aminoacyltransferases
  • sortase A
  • Lipase
  • 6-Phytase
  • Cellulase
  • Cysteine Endopeptidases