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Title (Primary) Evolutionary engineering of E. coli MG1655 for tolerance against isoprenol
Author Babel, H.; Krömer, J.O.;
Journal Biotechnology for Biofuels
Year 2020
Department SOMA;
Volume 13
Language englisch;
POF III (all) R61;
Supplements https://static-content.springer.com/esm/art%3A10.1186%2Fs13068-020-01825-6/MediaObjects/13068_2020_1825_MOESM1_ESM.pdf
Keywords Adaptive laboratory evolution; E. coli; Isoprenol; Butanol; Tolerance; Terpenes
Abstract Background:
Isoprenol is the basis for industrial flavor and vitamin synthesis and also a promising biofuel. Biotechnological production of isoprenol with E. coli is currently limited by the high toxicity of the final product. Adaptive laboratory evolution (ALE) is a promising method to address complex biological problems such as toxicity. Results:
Here we applied this method successfully to evolve E. coli towards higher tolerance against isoprenol, increasing growth at the half-maximal inhibitory concentration by 47%. Whole-genome re-sequencing of strains isolated from three replicate evolutions at seven time-points identified four major target genes for isoprenol tolerance: fabF, marC, yghB, and rob. We could show that knock-out of marC and expression of mutated Rob H(48) → frameshift increased tolerance against isoprenol and butanol. RNA-sequencing showed that the deletion identified upstream of yghB correlated with a strong overexpression of the gene. The knock-out of yghB demonstrated that it was essential for isoprenol tolerance. The mutated Rob protein and yghB deletion also lead to increased vanillin tolerance.
Conclusion:
Through ALE, novel targets for strain optimization in isoprenol production and also the production of other fuels, such as butanol, could be obtained. Their effectiveness could be shown through re-engineering. This paves the way for further optimization of E. coli for biofuel production.
ID 23839
Persistent UFZ Identifier https://www.ufz.de/index.php?en=20939&ufzPublicationIdentifier=23839
Babel, H., Krömer, J.O. (2020):
Evolutionary engineering of E. coli MG1655 for tolerance against isoprenol
Biotechnol. Biofuels 13 , art. 183