title: Improved production of membrane proteins in Escherichia coli by selective codon substitutions.
abstract: Membrane proteins are extremely challenging to produce in sufficient quantities for biochemical and structural analysis and there is a growing demand for solutions to this problem. In this study we attempted to improve expression of two difficult-to-express coding sequences (araH and narK) for membrane transporters. For both coding sequences, synonymous codon substitutions in the region adjacent to the AUG start led to significant improvements in expression, whereas multi-parameter sequence optimization of codons throughout the coding sequence failed. We conclude that coding sequences can be re-wired for high-level protein expression by selective engineering of the 5' coding sequence with synonymous codons, thus circumventing the need to consider whole sequence optimization.
authors.affiliation: Center for Biomembrane Research, Department of Biochemistry and Biophysics, Stockholm University, SE-106 91 Stockholm, Sweden. morno@biosustain.dtu.dk
authors.forename: Morten H H
authors.initials: MH
authors.lastname: Nørholm
authors.forename: Stephen
authors.initials: S
authors.lastname: Toddo
authors.forename: Minttu T I
authors.initials: MT
authors.lastname: Virkki
authors.forename: Sara
authors.initials: S
authors.lastname: Light
authors.forename: Gunnar
authors.initials: G
authors.lastname: von Heijne
authors.forename: Daniel O
authors.initials: DO
authors.lastname: Daley
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D003062:Codon; D029968:Escherichia coli Proteins / Q000096:biosynthesis* / Q000235:genetics; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid
chemical_list: D003062:Codon; D029968:Escherichia coli Proteins; D008565:Membrane Proteins
keywords: AraH; Codon optimization; Escherichia coli; Membrane protein; NarK; Over-expression; Synthetic coding sequence; Transporter