Development and characterization of SpyCatcher‐glycosyltransferase‐fusion proteins for highly specific automated glycan synthesis

Palm, Philip; Hußnätter, K.; Elling, Lothar

doi:10.1002/cite.202255302

Cited by 2 publications

(1 citation statement)

References 2 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…For this, microgels are synthesized on a microfluidic chip by utilizing water-in-oil droplets as microreactors for the crosslinking of poly(ethylene glycol) (PEG) prepolymers with counter-reactive vinyl sulfone (VS) and thiol (SH) groups. Recombinantly expressed SpyC GT fusion proteins 46 are immobilized onto these microgels using different methods, including enzyme encapsulation and postattachment. Various immobilization techniques, including noncovalent and covalent binding, are explored.…”

Section: Introductionmentioning

confidence: 99%

Microgels with Immobilized Glycosyltransferases for Enzymatic Glycan Synthesis

Sommerfeld,

Palm,

Hussnaetter

et al. 2024

Biomacromolecules

View full text Add to dashboard Cite

Glycans, composed of linked monosaccharides, play crucial roles in biology and find diverse applications. Enhancing their enzymatic synthesis can be achieved by immobilizing enzymes on materials such as microgels. Here, we present microgels with immobilized glycosyltransferases, synthesized through droplet microfluidics, immobilizing enzymes either via encapsulation or postattachment. SpyTag-SpyCatcher interaction was used for enzyme binding, among others. Fluorescamine and permeability assays confirmed enzyme immobilization and microgel porosity, while enzymatic activities were determined using HPLC. The potential application of microgels in cascade reactions involving multiple enzymes was demonstrated by combining β4GalT and α3GalT in an enzymatic reaction with high yields. Moreover, a cascade of β4GalT and β3GlcNAcT was successfully implemented. These results pave the way toward a modular membrane bioreactor for automated glycan synthesis containing the presented biocatalytic microgels.

show abstract

Section: Introductionmentioning

confidence: 99%