Improving cell-free glycoprotein synthesis by characterizing and enriching native membrane vesicles.

ABSTRACT: Cell-free gene expression (CFE) systems from crude cellular extracts have attracted much attention for biomanufacturing and synthetic biology. However, activating membrane-dependent functionality of cell-derived vesicles in bacterial CFE systems has been limited. Here, we address this limitation by characterizing native membrane vesicles in Escherichia coli-based CFE extracts and describing methods to enrich vesicles with heterologous, membrane-bound machinery. As a model, we focus on bacterial glycoengineering. We first use multiple, orthogonal techniques to characterize vesicles and show how extract processing methods can be used to increase concentrations of membrane vesicles in CFE systems. Then, we show that extracts enriched in vesicle number also display enhanced concentrations of h

SUBMITTER: Hershewe JM 

PROVIDER: S-EPMC8062659 | biostudies-literature | 2021 Apr

REPOSITORIES: biostudies-literature