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
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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
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Dynamic regulation of metabolic flux in engineered bacteria using a pathway-independent quorum-sensing circuit
TL;DR: A pathway-independent genetic control module is presented that can be used to dynamically regulate the expression of target genes and is applied to identify the optimal point to redirect glycolytic flux into heterologous engineered pathways in Escherichia coli, resulting in increased titers of myo-inositol and glucaric acid.
Journal ArticleDOI
Biosynthesis of cycloartenol by expression of plant and bacterial oxidosqualene cyclases in engineered Rhodobacter capsulatus.
Jennifer Hage-Hülsmann,Sabine Metzger,Vera Wewer,Felix Buechel,Katrin Troost,Katrin Troost,Stephan Thies,Stephan Thies,Anita Loeschcke,Anita Loeschcke,Karl-Erich Jaeger,Thomas Drepper +11 more
TL;DR: R. capsulatus is a promising alternative host for the functional expression of triterpene biosynthetic enzymes from plants and microbes and product titers can also be improved by suppression of competing precursor consumption.
Journal ArticleDOI
Synthetische Biologie - die Synthese der Biologie
Journal ArticleDOI
Improving the design of an oxidative stress sensing biosensor in yeast.
TL;DR: In this article, the authors proposed an approach to improve the design of an existing yeast TF-based biosensor in Saccharomyces cerevisiae based on the Yap1 transcription factor.
Journal ArticleDOI
Site-directed mutagenesis of the quorum-sensing transcriptional regulator SinR affects the biosynthesis of menaquinone in Bacillus subtilis.
Jing Wu,Wei Li,Zhao Shiguang,Qian Senhe,Zhou Wang,Meng-jie Zhou,Wen-song Hu,Jian Wang,Liu-xiu Hu,Yan Liu,Zheng-lian Xue +10 more
TL;DR: In this article, the authors used RNA-seq to analyze expression changes in the quorum sensing (QS) system, biofilm formation, and MK-7 synthesis pathway, which showed that the expressions of tapA, tasA and epsE were upregulated 9.79, 0.95, and 4.42-fold, respectively.
References
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Journal ArticleDOI
Production of the antimalarial drug precursor artemisinic acid in engineered yeast
Dae-Kyun Ro,Eric M. Paradise,Mario Ouellet,Karl Fisher,Karyn L. Newman,John M. Ndungu,Ho Kimberly,Eachus Rachel,Timothy S. Ham,James Kirby,Michelle C. Y. Chang,Sydnor T. Withers,Yoichiro Shiba,Richmond Sarpong,Jay D. Keasling +14 more
TL;DR: The engineering of Saccharomyces cerevisiae to produce high titres (up to 100 mg l-1) of artemisinic acid using an engineered mevalonate pathway, amorphadiene synthase, and a novel cytochrome P450 monooxygenase from A. annua that performs a three-step oxidation of amorpha-4,11-diene to art Artemisinic acid.
Journal ArticleDOI
Genesis: cluster analysis of microarray data
TL;DR: Genesis integrates various tools for microarray data analysis such as filters, normalization and visualization tools, distance measures as well as common clustering algorithms including hierarchical clustering, self-organizing maps, k-means, principal component analysis, and support vector machines.
Journal ArticleDOI
Engineering a mevalonate pathway in Escherichia coli for production of terpenoids
TL;DR: 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.
Journal ArticleDOI
Automated design of synthetic ribosome binding sites to control protein expression
TL;DR: A predictive method for designing synthetic ribosome binding sites is developed, enabling a rational control over the protein expression level, and is demonstrated by rationally optimizing protein expression to connect a genetic sensor to a synthetic circuit.
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