Modulation of Transaminase Activity by Encapsulation in Temperature-Sensitive Poly(N-acryloyl glycinamide) Hydrogels

Chembiochem. 2021 Dec 10;22(24):3452-3461. doi: 10.1002/cbic.202100427. Epub 2021 Oct 13.

Abstract

Smart hydrogels hold much potential for biocatalysis, not only for the immobilization of enzymes, but also for the control of enzyme activity. We investigated upper critical solution temperature-type poly N-acryloyl glycinamide (pNAGA) hydrogels as a smart matrix for the amine transaminase from Bacillus megaterium (BmTA). Physical entrapment of BmTA in pNAGA hydrogels results in high immobilization efficiency (>89 %) and high activity (97 %). The temperature-sensitiveness of pNAGA is preserved upon immobilization of BmTA and shows a gradual deswelling upon temperature reduction. While enzyme activity is mainly controlled by temperature, deactivation tended to be higher for immobilized BmTA (≈62-68 %) than for free BmTA (≈44 %), suggesting a deactivating effect due to deswelling of the pNAGA gel. Although the deactivation in response to hydrogel deswelling is not yet suitable for controlling enzyme activity sufficiently, it is nevertheless a good starting point for further optimization.

Keywords: activity regulation; biocatalysis; enzyme cascade; smart hydrogel; upper critical solution temperature.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acrylic Resins / chemistry
  • Acrylic Resins / metabolism*
  • Bacillus megaterium / enzymology*
  • Hydrogels / chemistry
  • Hydrogels / metabolism*
  • Molecular Structure
  • Temperature*
  • Transaminases / metabolism*

Substances

  • Acrylic Resins
  • Hydrogels
  • poly(N-acryloyl glycinamide)
  • Transaminases