bc9b00568_si_001.pdf (1.58 MB)
Selective Functionalization of Microgels with Enzymes by Sortagging
journal contribution
posted on 2019-10-11, 19:03 authored by Zhi Zou, Elisabeth Gau, Islam El-Awaad, Felix Jakob, Andrij Pich, Ulrich SchwanebergEnzyme 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.