A universal glycoenzyme biosynthesis pipeline that enables efficient cell-free remodeling of glycans.

ABSTRACT: The ability to reconstitute natural glycosylation pathways or prototype entirely new ones from scratch is hampered by the limited availability of functional glycoenzymes, many of which are membrane proteins that fail to express in heterologous hosts. Here, we describe a strategy for topologically converting membrane-bound glycosyltransferases (GTs) into water soluble biocatalysts, which are expressed at high levels in the cytoplasm of living cells with retention of biological activity. We demonstrate the universality of the approach through facile production of 98 difficult-to-express GTs, predominantly of human origin, across several commonly used expression platforms. Using a subset of these water-soluble enzymes, we perform structural remodeling of both free and protein-linked glycans i

SUBMITTER: Jaroentomeechai T 

PROVIDER: S-EPMC9592599 | biostudies-literature | 2022 Oct

REPOSITORIES: biostudies-literature