Details zur Publikation

Kategorie Textpublikation
Referenztyp Zeitschriften
DOI 10.1186/s13068-020-01825-6
Lizenz creative commons licence
Titel (primär) Evolutionary engineering of E. coli MG1655 for tolerance against isoprenol
Autor Babel, H.; Krömer, J.O.
Quelle Biotechnology for Biofuels
Erscheinungsjahr 2020
Department SOMA
Band/Volume 13
Seite von art. 183
Sprache englisch
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.
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.
dauerhafte UFZ-Verlinkung
Babel, H., Krömer, J.O. (2020):
Evolutionary engineering of E. coli MG1655 for tolerance against isoprenol
Biotechnol. Biofuels 13 , art. 183 10.1186/s13068-020-01825-6