Batianis, Christos and van Rosmalen, Rik P. and Fernandez, Pedro Monino and Labanaris, Konstantinos and Asin-Garcia, Enrique and Martin-Pascual, Maria and Jeschek, Markus and Weusthuis, Ruud A. and Suarez-Diez, Maria and Martins dos Santos, Vitor A. P. (2025) Computer-assisted multilevel optimization of malonyl-CoA availability in Pseudomonas putida. METABOLIC ENGINEERING, 90. pp. 165-177. ISSN 1096-7176, 1096-7184
Full text not available from this repository. (Request a copy)Abstract
Malonyl-CoA is the major precursor for the biosynthesis of diverse industrially valuable products such as fatty acids/alcohols, flavonoids, and polyketides. However, its intracellular availability is limited in most microbial hosts, hampering the industrial production of such chemicals. To address this limitation, we present a multilevel optimization workflow using modern metabolic engineering technologies to systematically increase the malonylCoA levels in Pseudomonas putida. The workflow involves the identification of gene downregulations, chassis selection, and optimization of the acetyl-CoA carboxylase complex through ribosome binding site engineering. Computational tools and high-throughput screening with a malonyl-CoA biosensor enabled the rapid evaluation of numerous genetic targets. Combining the most beneficial targets led to a 5.8-fold enhancement in the production titer of the valuable polyketide phloroglucinol. This study demonstrates the effective integration of computational and genetic technologies for engineering P. putida, opening new avenues for the development of industrially relevant strains and the investigation of fundamental biological questions.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | YARROWIA-LIPOLYTICA; ESCHERICHIA-COLI; TRANSFORMATION; PATHWAY; HOST; FLUX; Pseudomonas putida; Malonyl-CoA; Genome-scale modeling; CRISPRi; RBS optimization |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Mikrobiologie (Archaeenzentrum) |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 28 Jul 2026 06:02 |
| Last Modified: | 28 Jul 2026 06:02 |
| URI: | https://pred.uni-regensburg.de/id/eprint/67284 |
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