Journal ArticleDOI
Engineering dynamic pathway regulation using stress-response promoters
Robert H. Dahl,Fuzhong Zhang,Jorge Alonso-Gutierrez,Jorge Alonso-Gutierrez,Edward E. K. Baidoo,Edward E. K. Baidoo,Tanveer S. Batth,Tanveer S. Batth,Alyssa M. Redding-Johanson,Alyssa M. Redding-Johanson,Christopher J. Petzold,Christopher J. Petzold,Aindrila Mukhopadhyay,Aindrila Mukhopadhyay,Taek Soon Lee,Taek Soon Lee,Paul D. Adams,Paul D. Adams,Paul D. Adams,Jay D. Keasling +19 more
Reads0
Chats0
TLDR
This paper applied whole-genome transcript arrays to identify promoters that respond to the accumulation of toxic intermediates, and then used these promoters to control accumulation of the intermediate and improve the final titers of a desired product.Abstract:
Heterologous pathways used in metabolic engineering may produce intermediates toxic to the cell. Dynamic control of pathway enzymes could prevent the accumulation of these metabolites, but such a strategy requires sensors, which are largely unknown, that can detect and respond to the metabolite. Here we applied whole-genome transcript arrays to identify promoters that respond to the accumulation of toxic intermediates, and then used these promoters to control accumulation of the intermediate and improve the final titers of a desired product. We apply this approach to regulate farnesyl pyrophosphate (FPP) production in the isoprenoid biosynthetic pathway in Escherichia coli. This strategy improved production of amorphadiene, the final product, by twofold over that from inducible or constitutive promoters, eliminated the need for expensive inducers, reduced acetate accumulation and improved growth. We extended this approach to another toxic intermediate to demonstrate the broad utility of identifying novel sensor-regulator systems for dynamic regulation.read more
Citations
More filters
Journal ArticleDOI
Engineering Cellular Metabolism
TL;DR: How new technologies can enable metabolic engineering to be scaled up to the industrial level, either by cutting off the lines of control for endogenous metabolism or by infiltrating the system with disruptive, heterologous pathways that overcome cellular regulation is discussed.
Journal ArticleDOI
Principles of genetic circuit design
TL;DR: In this article, a review describes new tools that aid the construction of genetic circuits and discusses the failure modes encountered when assembling circuits, quantify their impact on performance, and review mitigation efforts.
Principles of genetic circuit design
TL;DR: Better tools, well-characterized parts and a comprehensive understanding of how to compose circuits are leading to a breakthrough in the ability to program living cells for advanced applications, from living therapeutics to the atomic manufacturing of functional materials.
Journal ArticleDOI
Improving fatty acids production by engineering dynamic pathway regulation and metabolic control
TL;DR: A genetically encoded metabolic switch that enables dynamic regulation of fatty acids (FA) biosynthesis in Escherichia coli was reported, able to dynamically compensate the critical enzymes involved in the supply and consumption of malonyl-CoA and efficiently redirect carbon flux toward FA biosynthesis.
Journal ArticleDOI
Metabolic Burden: Cornerstones in Synthetic Biology and Metabolic Engineering Applications
TL;DR: This work discusses cell physiological responses to metabolic burdens, as well as strategies to identify and resolve the carbon and energy burden problems, including metabolic balancing, enhancing respiration, dynamic regulatory systems, chromosomal engineering, decoupling cell growth with production phases, and co- utilization of nutrient resources.
References
More filters
Journal ArticleDOI
Mono and diterpene production in Escherichia coli
Keith Kinkead Reiling,Yasuo Yoshikuni,Vincent J. J. Martin,Jack D. Newman,Jörg Bohlmann,Jay D. Keasling,Jay D. Keasling +6 more
TL;DR: Construction and functional expression of GGPP and GPP operons provides an in vivo precursor platform host for the future engineering of di‐ and monoterpene cyclases and the overproduction of terpenes in bacteria.
Journal ArticleDOI
A model for improving microbial biofuel production using a synthetic feedback loop.
TL;DR: A model for cell growth and biofuel production is developed and used to compare several genetic control strategies for their ability to improve biofuel yields and shows that controlling efflux pump expression directly with a biofuel-responsive promoter is a straightforward way of improving bio fuel production.
Journal ArticleDOI
SNOMAD (Standardization and NOrmalization of MicroArray Data): web-accessible gene expression data analysis
TL;DR: SNOMAD is a collection of algorithms for the normalization and standardization of gene expression datasets derived from diverse biological and technological sources which correct for bias and variance which are non-uniformly distributed across the range of microarray element signal intensities.
Journal ArticleDOI
Cell-Specific and Conditional Expression of Caffeoyl-Coenzyme A-3-O-Methyltransferase in Poplar
Cuiying Chen,Hugo Meyermans,Bart Burggraeve,Riet De Rycke,Kentaro Inoue,Vera De Vleesschauwer,Marijke Steenackers,Marc Van Montagu,Gilbert Engler,Wout Boerjan +9 more
TL;DR: The expression pattern conferred by the two CCoAOMT promoters when fused to the gus-coding sequence in transgenic poplar is described and may explain the heterogeneity in lignin composition that is observed between cell types and upon different environmental conditions.
Journal ArticleDOI
The in vivo synthesis of plant sesquiterpenes by Escherichia coli.
TL;DR: It is concluded that the limiting factor for sesquiterpene synthesis in E. coli is the poor expression of the cyclase enzyme and not supply of the FDP precursor.
Related Papers (5)
Design of a dynamic sensor-regulator system for production of chemicals and fuels derived from fatty acids.
Improving lycopene production in Escherichia coli by engineering metabolic control
William R. Farmer,James C. Liao +1 more