Bicistronic Design-Based Continuous and High-Level Membrane Protein Production in Escherichia coli

Nico J. Claassens*, Max Finger-Bou, Bart Scholten, Frederieke Muis, Jonas J. De Groot, Jan Willem De Gier, Willem M. De Vos, John Van Der Oost

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

17 Citations (Scopus)


Escherichia coli has been widely used as a platform microorganism for both membrane protein production and cell factory engineering. The current methods to produce membrane proteins in this organism require the induction of target gene expression and often result in unstable, low yields. Here, we present a method combining a constitutive promoter with a library of bicistronic design (BCD) elements, which enables inducer-free, tuned translation initiation for optimal protein production. Our system mediates stable, constitutive production of bacterial membrane proteins at yields that outperform those obtained with E. coli Lemo21(DE3), the current gold standard for bacterial membrane protein production. We envisage that the continuous, fine-tunable, and high-level production of membrane proteins by our method will greatly facilitate their study and their utilization in engineering cell factories.

Original languageEnglish
Pages (from-to)1685-1690
Number of pages6
JournalACS synthetic biology
Issue number7
Publication statusPublished - 17 Jun 2019


  • bicistronic design
  • Escherichia coli
  • membrane proteins
  • protein production
  • translational coupling


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