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Inverse Metabolic Engineering for Enhanced Glycoprotein Production in Escherichia coli
Stephen R P Jaffé1, Benjamin Strutton, Jagroop Pandhal
1Department of Chemical and Biological Engineering, ChELSI Institute, University of Sheffield, Sheffield, UK.
Abstract:
Inverse metabolic engineering (IME) provides a strategy to rapidly identify the genetic elements responsible for the desired phenotype of a chosen target organism. This methodology has been successfully applied towards enhancing the N-linked glycosylation efficiency of Escherichia coli. Here, we describe the generation of differentially sized libraries from the E. coli W3110 genome followed by high-throughput semiquantitative glycan specific screening. DNA sequenced targets demonstrating increased levels of glycan production were selected for forward engineering, protein overexpression, and absolute quantification of glycoproteins.
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