Fig. 1 | Nature Communications

Fig. 1

From: Single-pot glycoprotein biosynthesis using a cell-free transcription-translation system enriched with glycosylation machinery

Fig. 1

Schematic of single-pot CFGpS technology. Glycoengineered E. coli that are modified with (i) genomic mutations that benefit glycosylation reactions and (ii) plasmid DNA for producing essential glycosylation components (i.e., OSTs, LLOs) serve as the source strain for producing crude S30 extracts. Candidate glycosylation components can be derived from all kingdoms of life and include single-subunit OSTs like C. jejuni PglB and LLOs bearing N-glycans from C. jejuni that are assembled on Und-PP by the Pgl pathway enzymes. Following extract preparation by lysis of the source strain, one-pot biosynthesis of N-glycoproteins is initiated by priming the extract with DNA encoding the acceptor protein of interest

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