Using promoter libraries to reduce metabolic burden due to plasmid-encoded proteins in recombinant Escherichia coli

New Biotechnology
Martina PasiniPau Ferrer

Abstract

The over-expression of proteins in recombinant host cells often requires a significant amount of resources causing an increase in the metabolic load for the host. This results in a variety of physiological responses leading to altered growth parameters, including growth inhibition or activation of secondary metabolism pathways. Moreover, the expression of other plasmid-encoded genes such as antibiotic resistance genes or repressor proteins may also alter growth kinetics. In this work, we have developed a second-generation system suitable for Escherichia coli expression with an antibiotic-free plasmid maintenance mechanism based on a glycine auxotrophic marker (glyA). Metabolic burden related to plasmid maintenance and heterologous protein expression was minimized by tuning the expression levels of the repressor protein (LacI) and glyA using a library of promoters and applying synthetic biology tools that allow the rapid construction of vectors. The engineered antibiotic-free expression system was applied to the l-fuculose phosphate aldolase (FucA) over-production, showing an increase in production up to 3.8-fold in terms of FucA yield (mgg(-1)DCW) and 4.5-fold in terms of FucA activity (AUg(-1)DCW) compared to previous expressi...Continue Reading

References

Mar 1, 1996·Trends in Biotechnology·S Y Lee
Oct 6, 1999·Current Opinion in Biotechnology·F Baneyx
Nov 27, 2002·Proceedings of the National Academy of Sciences of the United States of America·Yohei YokobayashiFrances H Arnold
Jan 1, 1995·Biotechnology Advances·B R Glick
Jan 7, 2005·Applied Microbiology and Biotechnology·S Jana, J K Deb
Sep 8, 2005·Microbial Cell Factories·Jacob Glenting, Stephen Wessels
Sep 19, 2006·Current Opinion in Biotechnology·Christopher A Voigt
Nov 6, 2008·PloS One·Carola EnglerSylvestre Marillonnet
Apr 17, 2010·Applied Microbiology and Biotechnology·Heinz Neumann, Petra Neumann-Staubitz
May 17, 2011·Microbial Cell Factories·Tomohiro MakinoGeorge Georgiou
Oct 29, 2011·Applied Microbiology and Biotechnology·Jacob E VickClaudia Schmidt-Dannert

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Citations

Oct 19, 2016·Current Opinion in Microbiology·Olivier BorkowskiTom Ellis
Apr 14, 2016·Biotechnology Progress·Desmond M SchofieldDarren N Nesbeth
Feb 6, 2018·Nature Reviews. Microbiology·David T Riglar, Pamela A Silver
Mar 27, 2018·Nature Methods·Francesca CeroniTom Ellis
Mar 24, 2018·Journal of Biological Engineering·José-Aníbal Mora-Villalobos, An-Ping Zeng
Oct 6, 2018·Nucleic Acids Research·Shuyi Zhang, Christopher A Voigt
Mar 3, 2020·Frontiers in Microbiology·Hui LiMichael G Gänzle

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