ArticleACS synthetic biology2017
Refactoring the Embden-Meyerhof-Parnas Pathway as a Whole of Portable GlucoBricks for Implantation of Glycolytic Modules in Gram-Negative Bacteria.
Article in ACS synthetic biology, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 25 papers.
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25 citing papers in PubMed, 66 citations in OpenAlex.
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- Reconfiguration of metabolic fluxes in Pseudomonas putida as a response to sub-lethal oxidative stress.The ISME journal · 2021Article
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- The Inactivation of Enzymes Belonging to the Central Carbon Metabolism Is a Novel Mechanism of Developing Antibiotic Resistance.mSystems · 2020Article
- Synthetic control of plasmid replication enables target- and self-curing of vectors and expedites genome engineering ofMetabolic engineering communications · 2020Article
- Chromosome-free bacterial cells are safe and programmable platforms for synthetic biology.Proceedings of the National Academy of Sciences of the United States of America · 2020Article
- Engineering sucrose metabolism in Pseudomonas putida highlights the importance of porins.Microbial biotechnology · 2020Article
- Gross transcriptomic analysis of Pseudomonas putida for diagnosing environmental shifts.Microbial biotechnology · 2020Article
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The Embden-Meyerhof-Parnas (EMP) pathway is generally considered to be the biochemical standard for glucose catabolism. Alas, its native genomic organization and the control of gene expression in Escherichia coli are both very intricate, which limits the portability of the EMP pathway to other biotechnologically important bacterial hosts that lack the route. In this work, the genes encoding all the enzymes of the linear EMP route have been individually recruited from the genome of E. coli K-12, edited in silico to remove their endogenous regulatory signals, and synthesized de novo following a standard (GlucoBrick) that enables their grouping in the form of functional modules at the user's will. After verifying their activity in several glycolytic mutants of E. coli, the versatility of these GlucoBricks was demonstrated in quantitative physiology tests and biochemical assays carried out in Pseudomonas putida KT2440 and P. aeruginosa PAO1 as the heterologous hosts. Specific configurations of GlucoBricks were also adopted to streamline the downward circulation of carbon from hexoses to pyruvate in E. coli recombinants, thereby resulting in a 3-fold increase of poly(3-hydroxybutyrate) synthesis from glucose. Refactoring whole metabolic blocks in the fashion described in this work thus eases the engineering of biochemical processes where the optimization of carbon traffic is facilitated by the operation of the EMP pathway-which yields more ATP than other glycolytic routes such as the Entner-Doudoroff pathway.
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