A high-throughput screen for hyaluronic acid accumulation in recombinant Escherichia coli transformed by libraries of engineered sigma factors

2008 ◽  
Vol 101 (4) ◽  
pp. 788-796 ◽  
Author(s):  
Huimin Yu ◽  
Keith Tyo ◽  
Hal Alper ◽  
Daniel Klein-Marcuschamer ◽  
Gregory Stephanopoulos
Keyword(s):  
Escherichia Coli ◽  
Hyaluronic Acid ◽  
High Throughput ◽  
Sigma Factors ◽  
Recombinant Escherichia Coli ◽  
High Throughput Screen ◽  
Acid Accumulation

scholarly journals Melanin-Based High-Throughput Screen for l-Tyrosine Production in Escherichia coli

2007 ◽  
Vol 74 (4) ◽  
pp. 1190-1197 ◽  
Author(s):  
Christine Nicole S. Santos ◽  
Gregory Stephanopoulos
Keyword(s):  
Escherichia Coli ◽  
High Throughput ◽  
Combinatorial Libraries ◽  
Selection Strategy ◽  
High Throughput Screen ◽  
Bacterial Strains ◽  
Gene Encoding ◽  
Strong Linear Correlation ◽  
Low Levels ◽  
Tyrosine Content

ABSTRACT We present the development of a simple, high-throughput screen for identifying bacterial strains capable of l-tyrosine production. Through the introduction of a heterologous gene encoding a tyrosinase, we were able to link l-tyrosine production in Escherichia coli with the synthesis of the black and diffusible pigment melanin. Although melanin was initially produced only at low levels in morpholinepropanesulfonic acid (MOPS) minimal medium, phosphate supplementation was found to be sufficient for increasing both the rates of synthesis and the final titers of melanin. Furthermore, a strong linear correlation between extracellular l-tyrosine content and melanin formation was observed by use of this new medium formulation. A selection strategy that utilizes these findings has been developed and has been shown to be effective in screening large combinatorial libraries in the search for l-tyrosine-overproducing strains.

scholarly journals A High-Throughput Screen of the GTPase Activity of Escherichia coli EngA to Find an Inhibitor of Bacterial Ribosome Biogenesis

2013 ◽  
Vol 18 (7) ◽  
pp. 830-836 ◽  
Author(s):  
Amrita Bharat ◽  
Jan E. Blanchard ◽  
Eric D. Brown
Keyword(s):  
Escherichia Coli ◽  
High Throughput ◽  
High Throughput Screening ◽  
Ribosome Biogenesis ◽  
Signal Amplitude ◽  
Low Noise ◽  
High Throughput Screen ◽  
Reaction Conditions ◽  
Screening Process ◽  
Bacterial Ribosome

The synthesis of ribosomes is an essential process, which is aided by a variety of trans-acting factors in bacteria. Among these is a group of GTPases essential for bacterial viability and emerging as promising targets for new antibacterial agents. Herein, we describe a robust high-throughput screening process for inhibitors of one such GTPase, the Escherichia coli EngA protein. The primary screen employed an assay of phosphate production in a 384-well density. Reaction conditions were chosen to maximize sensitivity for the discovery of competitive inhibitors while maintaining a strong signal amplitude and low noise. In a pilot screen of 31,800 chemical compounds, 44 active compounds were identified. Furthermore, we describe the elimination of nonspecific inhibitors that were detergent sensitive or reactive as well as those that interfered with the high-throughput phosphate assay. Four inhibitors survived these common counterscreens for nonspecificity, but these chemicals were also inhibitors of the unrelated enzyme dihydrofolate reductase, suggesting that they too were promiscuously active. The high-throughput screen of the EngA protein described here provides a meticulous pilot study in the search for specific inhibitors of GTPases involved in ribosome biogenesis.

scholarly journals High-Throughput Screen Identifying the Thiosemicarbazone NSC319726 Compound as a Potent Antimicrobial Lead Against Resistant Strains of Escherichia coli

Biomolecules ◽  
2018 ◽  
Vol 8 (4) ◽  
pp. 166 ◽  
Author(s):  
Carmen Sadaka ◽  
Peter Damborg ◽  
Jeffrey L. Watts
Keyword(s):  
Escherichia Coli ◽  
High Throughput ◽  
Multidrug Resistant ◽  
Coli Strain ◽  
High Throughput Screen ◽  
Lead Discovery ◽  
E Coli ◽  
Resistant Strains ◽  
Susceptibility Profiles ◽  
Antibiotic Discovery

Antibiotic discovery is vital when considering the increasing antimicrobial resistance threat. The aim of this work was to provide a high-throughput screen (HTS) assay using multidrug-resistant Escherichia coli strains to enable further research into antimicrobial lead discovery and identify novel antimicrobials. This study describes a primary HTS of a diverse library of 7884 small molecules against a susceptible E. coli strain. A secondary screening of 112 molecules against four E. coli strains with different susceptibility profiles revealed NSC319726 as a potential antimicrobial lead serving as a novel template. NSC319726 is a good candidate for an analoguing program.

Metabolic profiling of recombinant Escherichia coli cultivations based on high-throughput FT-MIR spectroscopic analysis

2016 ◽  
Vol 33 (2) ◽  
pp. 285-298 ◽  
Author(s):  
Kevin C. Sales ◽  
Filipa Rosa ◽  
Bernardo R. Cunha ◽  
Pedro N. Sampaio ◽  
Marta B. Lopes ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
High Throughput ◽  
Metabolic Profiling ◽  
Spectroscopic Analysis ◽  
Recombinant Escherichia Coli

scholarly journals High Throughput Screen for Escherichia coli Twin Arginine Translocation (Tat) Inhibitors

PLoS ONE ◽  
2016 ◽  
Vol 11 (2) ◽  
pp. e0149659 ◽  
Author(s):  
Umesh K. Bageshwar ◽  
Lynn VerPlank ◽  
Dwight Baker ◽  
Wen Dong ◽  
Shruthi Hamsanathan ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
High Throughput ◽  
High Throughput Screen ◽  
Twin Arginine Translocation
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