Evolutionary Engineering of E. Coli MG1655 for Tolerance Against Isoprenol

Overview

Journal Biotechnol Biofuels

Publisher Biomed Central

Specialty Biotechnology

Date 2020 Dec 9

PMID 33292484

Citations 5

Authors

Heiko Babel

Jens O Kromer

Affiliations

Soon will be listed here.

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.

Citing Articles

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Enhancement of linalool production in Saccharomyces cerevisiae by utilizing isopentenol utilization pathway.

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Isopentenol Utilization Pathway for the Production of Linalool in Escherichia coli Using an Improved Bacterial Linalool/Nerolidol Synthase.

Ferraz C, Leferink N, Kosov I, Scrutton N Chembiochem. 2021; 22(13):2325-2334.

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Machas M, Kurgan G, Abed O, Shapiro A, Wang X, Nielsen D J Ind Microbiol Biotechnol. 2021; 48(1-2).

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