Improving heterologous membrane protein production in Escherichia coli by combining transcriptional tuning and codon usage algorithms.

Nico J. Claassens*, Melvin F. Siliakus, Sebastiaan K. Spaans, Sjoerd C.A. Creutzburg, Bart Nijsse, Peter J. Schaap, Tessa Quax, John van der Oost

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

32 Citations (Scopus)
41 Downloads (Pure)

Abstract

High-level, recombinant production of membrane-integrated proteins in Escherichia coli is extremely relevant for many purposes, but has also been proven challenging. Here we study a combination of transcriptional fine-tuning in E. coli LEMO21(DE3) with different codon usage algorithms for heterologous production of membrane proteins. The overex- pression of 6 different membrane proteins is compared for the wild-type gene codon usage variant, a commercially codon-optimized variant, and a codon-harmonized variant. We show that transcriptional fine-tuning plays a major role in improving the production of all tested proteins. Moreover, different codon usage variants significantly improved production of some of the tested proteins. However, not a single algorithm performed consistently best for the membrane-integrated production of the 6 tested proteins. In conclusion, for improving heterologous membrane protein production in E. coli, the major effect is accomplished by transcriptional tuning. In addition, further improvements may be realized by attempting dif- ferent codon usage variants, such as codon harmonized variants, which can now be easily generated through our online Codon Harmonizer tool.
Original languageEnglish
Article numbere0184355
Number of pages17
JournalPLoS ONE
Volume12
Issue number9
DOIs
Publication statusPublished - 2017

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