A Recoverable Enzymatic Microgel Based on Biomolecular Recognition

The enzyme β-galactosidase has been immobilized through incorporation into a selectively soluble microgel, prepared from DNA, biotinylated peptide nucleic acid (PNA), and the protein avidin. The enzyme was conjugated to avidin, allowing it to be integrated directly into the microgel network. Efficient hydrolysis of a small-molecule substrate occurred at 37°, but cooling and centrifuging led to precipitation of the microgels and product separation. The microgels were then reconstituted by adding fresh buffer and shaking. The enzymatic activity was completely recovered through repeated cycles. This method should be generalizable to a wide variety of other enzymes and substrates.