| Title | System metabolic engineering of Escherichia coli W for the production of 2-ketoisovalerate using unconventional feedstock |
|---|---|
| ID_Doc | 24821 |
| Authors | Carranza-Saavedra, D; Torres-Bacete, J; Blázquez, B; Henao, CPS; Montoya, JEZ; Nogales, J |
| Title | System metabolic engineering of Escherichia coli W for the production of 2-ketoisovalerate using unconventional feedstock |
| Year | 2023 |
| Published | |
| Abstract | Replacing traditional substrates in industrial bioprocesses to advance the sustainable production of chemicals is an urgent need in the context of the circular economy. However, since the limited degradability of non-conventional carbon sources often returns lower yields, effective exploitation of such substrates requires a multi-layer optimization which includes not only the provision of a suitable feedstock but the use of highly robust and metabolically versatile microbial biocatalysts. We tackled this challenge by means of systems metabolic engineering and validated Escherichia coli W as a promising cell factory for the production of the key building block chemical 2-ketoisovalerate (2-KIV) using whey as carbon source, a widely available and low-cost agro-industrial waste. First, we assessed the growth performance of Escherichia coli W on mono and disaccharides and demonstrated that using whey as carbon source enhances it significantly. Second, we searched the available literature and used metabolic modeling approaches to scrutinize the metabolic space of E. coli and explore its potential for overproduction of 2-KIV identifying as basic strategies the block of pyruvate depletion and the modulation of NAD/NADP ratio. We then used our model predictions to construct a suitable microbial chassis capable of overproducing 2-KIV with minimal genetic perturbations, i.e., deleting the pyruvate dehydrogenase and malate dehydrogenase. Finally, we used modular cloning to construct a synthetic 2-KIV pathway that was not sensitive to negative feedback, which effectively resulted in a rerouting of pyruvate towards 2-KIV. The resulting strain shows titers of up to 3.22 +/- 0.07 g/L of 2-KIV and 1.40 +/- 0.04 g/L of L-valine in 24 h using whey in batch cultures. Additionally, we obtained yields of up to 0.81 g 2-KIV/g substrate. The optimal microbial chassis we present here has minimal genetic modifications and is free of nutritional autotrophies to deliver high 2-KIV production rates using whey as a non-conventional substrate. |
| https://www.frontiersin.org/articles/10.3389/fbioe.2023.1176445/pdf |
Similar Articles
| ID | Score | Article |
|---|---|---|
22414
|
Kim, S; Giraldo, N; Rainaldi, V; Machens, F; Collas, F; Kubis, A; Kensy, F; Bar-Even, A; Lindner, SN Optimizing E. coli as a formatotrophic platform for bioproduction via the reductive glycine pathway(2023) | |
| 14813
|
Lyu, X; Nuhu, M; Candry, P; Wolfanger, J; Betenbaugh, M; Saldivar, A; Zuniga, C; Wang, Y; Shrestha, S Top-down and bottom-up microbiome engineering approaches to enable biomanufacturing from waste biomass(2024) | |
| 29424
|
Li, Y; Cheng, ZZ; Zhao, CL; Gao, C; Song, W; Liu, LM; Chen, XL Reprogramming Escherichia coli Metabolism for Bioplastics Synthesis from Waste Cooking Oil(2021)Acs Synthetic Biology, 10.0, 8 | |
| 12792
|
Gao, ZX; Guo, SQ; Chen, YH; Chen, HS; Fu, RZ; Song, QQ; Li, S; Lou, WY; Fan, DD; Li, Y; Yang, SH; Gonzalez, R; Fei, Q A novel nutritional induction strategy flexibly switching the biosynthesis of food-like products from methane by a methanotrophic bacterium(2024)Green Chemistry, 26.0, 12 | |
| 5285
|
Intasian, P; Prakinee, K; Phintha, A; Trisrivirat, D; Weeranoppanant, N; Wongnate, T; Chaiyen, P Enzymes, In Vivo Biocatalysis, and Metabolic Engineering for Enabling a Circular Economy and Sustainability(2021)Chemical Reviews, 121, 17 | |
| 13839
|
Baptista, SL; Costa, CE; Cunha, JT; Soares, PO; Domingues, L Metabolic engineering of Saccharomyces cerevisiae for the production of top value chemicals from biorefinery carbohydrates(2021) | |
| 20637
|
Joshi, S; Mishra, S Recent advances in biofuel production through metabolic engineering(2022) | |
| 15472
|
Wegat, V; Fabarius, JT; Sieber, V Synthetic methylotrophic yeasts for the sustainable fuel and chemical production(2022)Biotechnology For Biofuels And Bioproducts, 15, 1 | |
| 14051
|
Bae, J; Park, C; Jung, H; Jin, S; Cho, BK Harnessing acetogenic bacteria for one-carbon valorization toward sustainable chemical production(2024)Rsc Chemical Biology, 5, 9 | |
| 7873
|
Lasagni, F; Cassanelli, S; Gullo, M How carbon sources drive cellulose synthesis in two Komagataeibacter xylinus strains(2024)Scientific Reports, 14, 1 |