Synthetic protein scaffolds provide modular control over metabolic flux

  title={Synthetic protein scaffolds provide modular control over metabolic flux},
  author={John E. Dueber and Gabriel C. Wu and G Reza Malmirchegini and Tae Seok Moon and Christopher J. Petzold and Adeeti V. Ullal and Kristala Jones Prather and Jay D. Keasling},
  journal={Nature Biotechnology},
Engineered metabolic pathways constructed from enzymes heterologous to the production host often suffer from flux imbalances, as they typically lack the regulatory mechanisms characteristic of natural metabolism. [] Key Result The natural modularity of these domains enabled us to optimize the stoichiometry of three mevalonate biosynthetic enzymes recruited to a synthetic complex and thereby achieve 77-fold improvement in product titer with low enzyme expression and reduced metabolic load.

Spatial organization of enzymes for metabolic engineering.

Modular enzyme assembly for enhanced cascade biocatalysis and metabolic flux

A pair of short peptide tags (RIAD and RIDD) are chosen to create scaffold-free enzyme assemblies and synthetic metabolic nodes to impose metabolic control in biosynthetic microbe factories.

Integrating the protein and metabolic engineering toolkits for next-generation chemical biosynthesis.

The importance of protein engineering for advancing metabolic engineering of secondary metabolism pathways through integrated enzyme optimization can enhance the collective toolkit of microbial engineering to shape the future of chemical manufacturing.

[Research progress on synthetic scaffold in metabolic engineering - a review].

The application of synthetic scaffolds in metabolic engineering is summarized, the main principle of scaffold designs are outlined, and the current challenges in their application are highlighted.

Protein design for pathway engineering.

Customized optimization of metabolic pathways by combinatorial transcriptional engineering

This work reports a simple, efficient and programmable approach named ‘customized optimization of metabolic pathways by combinatorial transcriptional engineering (COMPACTER)’ for rapid tuning of gene expression in a heterologous pathway under distinct metabolic backgrounds.

Constructing de novo biosynthetic pathways for chemical synthesis inside living cells.

Efforts to understand the relationship among sequence, structure, and function in the basic biochemical sciences can advance these goals for synthetic biology applications while also serving as an experimental platform for elucidating the in vivo specificity and function of enzymes and reconstituting complex biochemical traits for study in a living model organism.

Light-based control of metabolic flux through assembly of synthetic organelles

It is reported that optogenetic control can be used to extend compartmentalization and dynamic control to engineered metabolisms in yeast and enhances product formation and specificity during deoxyviolacein biosynthesis by decreasing concentrations of intermediate metabolites and reducing flux through competing pathways.

Synthetic scaffolds for pathway enhancement.




Application of Functional Genomics to Pathway Optimization for Increased Isoprenoid Production

The results indicate that HMG-CoA inhibits fatty acid biosynthesis in the microbial host, leading to generalized membrane stress, demonstrating the utility of using transcriptomic and metabolomic methods to optimize synthetic biological systems.

Engineering the spatial organization of metabolic enzymes: mimicking nature's synergy.

Metabolic fluxes and metabolic engineering.

It is shown how metabolic fluxes can be used in the systematic elucidation of metabolic control in the framework of reaction grouping and top-down metabolic control analysis.

Using Engineered Scaffold Interactions to Reshape MAP Kinase Pathway Signaling Dynamics

It is demonstrated that the Ste5 scaffold protein can be used as a platform to systematically reshape output of the yeast mating MAP kinase pathway, and synthetic positive- and negative-feedback loops are constructed.

Metabolic engineering for drug discovery and development

The impact that converging developments in genetic engineering and biosynthetic chemistry are having on natural-product drug discovery is reviewed.

Combinatorial engineering of intergenic regions in operons tunes expression of multiple genes

A method for tuning the expression of multiple genes within operons by generating libraries of tunable intergenic regions (TIGRs), recombining various post-transcriptional control elements and screening for the desired relative expression levels is described.

Production of Glucaric Acid from a Synthetic Pathway in Recombinant Escherichia coli

A synthetic pathway has been constructed for the production of glucuronic and glucaric acids from glucose in Escherichia coli and the inclusion of a third enzyme, uronate dehydrogenase (Udh) from Pseudomonas syringae, facilitated the conversion ofglucuronic acid to glucAric acid.

Engineering a mevalonate pathway in Escherichia coli for production of terpenoids

The strains developed in this study can serve as platform hosts for the production of any terpenoid compound for which a terpene synthase gene is available, and are the universal precursors to all isoprenoids.

A synthetic oscillatory network of transcriptional regulators

This work used three transcriptional repressor systems that are not part of any natural biological clock to build an oscillating network, termed the repressilator, in Escherichia coli, which periodically induces the synthesis of green fluorescent protein as a readout of its state in individual cells.