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

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

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.

LanguageEnglish
Pages1685-1690
Number of pages6
JournalACS synthetic biology
Volume8
Issue number7
DOIs
Publication statusPublished - 17 Jun 2019

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Escherichia coli
Membrane Proteins
Proteins
Membranes
Cell Engineering
Bacterial Proteins
Industrial plants
Translational Peptide Chain Initiation
Cell engineering
Gene expression
Microorganisms
Libraries
Gene Expression

Keywords

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

Cite this

Claassens, Nico J. ; Finger-Bou, Max ; Scholten, Bart ; Muis, Frederieke ; De Groot, Jonas J. ; De Gier, Jan Willem ; De Vos, Willem M. ; Van Der Oost, John. / Bicistronic Design-Based Continuous and High-Level Membrane Protein Production in Escherichia coli. In: ACS synthetic biology. 2019 ; Vol. 8, No. 7. pp. 1685-1690.
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Bicistronic Design-Based Continuous and High-Level Membrane Protein Production in Escherichia coli. / Claassens, Nico J.; Finger-Bou, Max; Scholten, Bart; Muis, Frederieke; De Groot, Jonas J.; De Gier, Jan Willem; De Vos, Willem M.; Van Der Oost, John.

In: ACS synthetic biology, Vol. 8, No. 7, 17.06.2019, p. 1685-1690.

Research output: Contribution to journalArticleAcademicpeer-review

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AU - Claassens, Nico J.

AU - Finger-Bou, Max

AU - Scholten, Bart

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AU - De Groot, Jonas J.

AU - De Gier, Jan Willem

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