Enzymatic Azido-GalNAc-Functionalized Silk Fibroin for Click Chemistry Conjugation.

Silk is a popular protein biomaterial that has been used for various purposes such as tissue scaffolding, textiles and hydrogels. Various methods for covalent conjugation of functional molecules such as small molecule sensors and enzymes have been developed to create functionalized silk biomaterials. Here, we report a method for silk functionalization by using -GalNAc-transferases and azide-modified UDP-GalNAc nucleotide sugar substrates to incorporate azide functional groups onto the silk fibroin protein for functionalization with cycloalkynes by click chemistry. Using ppGalNAc-T1 and T13 enzymes, we could transfer azide-modified GalNAc monosaccharides onto fibroin and as a proof of concept, conjugated a strain-alkyne-functionalized Cy5 fluorophore to produce a Cy5-conjugated fibroin hydrogel. This facile azido functionalization of the silk has the potential for attachment of any cycloalkyne moiety.