title: Inverse Metabolic Engineering for Enhanced Glycoprotein Production in Escherichia coli.
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.
authors.affiliation: Department of Chemical and Biological Engineering, ChELSI Institute, University of Sheffield, Sheffield, UK.
authors.forename: Stephen R P
authors.initials: SR
authors.lastname: Jaffé
authors.forename: Benjamin
authors.initials: B
authors.lastname: Strutton
authors.forename: Jagroop
authors.initials: J
authors.lastname: Pandhal
authors.forename: Phillip C
authors.initials: PC
authors.lastname: Wright
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D016678:Genome / Q000235:genetics; D006023:Glycoproteins / Q000235:genetics* / Q000378:metabolism*; D006031:Glycosylation; D060847:Metabolic Engineering / Q000379:methods; D011134:Polysaccharides / Q000235:genetics / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D006023:Glycoproteins; D011134:Polysaccharides