Improving heterologous membrane protein production in Escherichia coli by combining transcriptional tuning and codon usage algorithms.
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 heterologo...
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2017-01-01
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| Series: | PLoS ONE |
| Online Access: | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0184355&type=printable |
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| author | Nico J Claassens Melvin F Siliakus Sebastiaan K Spaans Sjoerd C A Creutzburg Bart Nijsse Peter J Schaap Tessa E F Quax John van der Oost |
| author_facet | Nico J Claassens Melvin F Siliakus Sebastiaan K Spaans Sjoerd C A Creutzburg Bart Nijsse Peter J Schaap Tessa E F Quax John van der Oost |
| author_sort | Nico J Claassens |
| collection | DOAJ |
| description | 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 overexpression 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 different codon usage variants, such as codon harmonized variants, which can now be easily generated through our online Codon Harmonizer tool. |
| format | Article |
| id | doaj-art-c963d8bea3bf4f098f799476ca6e0702 |
| institution | OA Journals |
| issn | 1932-6203 |
| language | English |
| publishDate | 2017-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj-art-c963d8bea3bf4f098f799476ca6e07022025-08-20T02:04:20ZengPublic Library of Science (PLoS)PLoS ONE1932-62032017-01-01129e018435510.1371/journal.pone.0184355Improving heterologous membrane protein production in Escherichia coli by combining transcriptional tuning and codon usage algorithms.Nico J ClaassensMelvin F SiliakusSebastiaan K SpaansSjoerd C A CreutzburgBart NijssePeter J SchaapTessa E F QuaxJohn van der OostHigh-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 overexpression 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 different codon usage variants, such as codon harmonized variants, which can now be easily generated through our online Codon Harmonizer tool.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0184355&type=printable |
| spellingShingle | Nico J Claassens Melvin F Siliakus Sebastiaan K Spaans Sjoerd C A Creutzburg Bart Nijsse Peter J Schaap Tessa E F Quax John van der Oost Improving heterologous membrane protein production in Escherichia coli by combining transcriptional tuning and codon usage algorithms. PLoS ONE |
| title | Improving heterologous membrane protein production in Escherichia coli by combining transcriptional tuning and codon usage algorithms. |
| title_full | Improving heterologous membrane protein production in Escherichia coli by combining transcriptional tuning and codon usage algorithms. |
| title_fullStr | Improving heterologous membrane protein production in Escherichia coli by combining transcriptional tuning and codon usage algorithms. |
| title_full_unstemmed | Improving heterologous membrane protein production in Escherichia coli by combining transcriptional tuning and codon usage algorithms. |
| title_short | Improving heterologous membrane protein production in Escherichia coli by combining transcriptional tuning and codon usage algorithms. |
| title_sort | improving heterologous membrane protein production in escherichia coli by combining transcriptional tuning and codon usage algorithms |
| url | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0184355&type=printable |
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