Design Space Exploration of the Violacein Pathway in Escherichia coli Based Cell-Free System

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Title Design Space Exploration of the Violacein Pathway in Escherichia coli Based Cell-Free System
Authors Phuc H.B. Nguyen, Yong Wu, Shaobin Guo, Richard M Murray
Source Submitted, 2016 Synthetic Biology: Engineering, Evolution and Design (SEED) Conference
Abstract In this study, an Escherichia coli (E. coli) based transcription translation cell-free system (TX-TL) was employed to sample various enzyme expression levels of the violacein pathway. TX-TL enables rapid modifications and prototyping of the pathway without complicated cloning cycles. The violacein metabolic pathway has been successfully reconstructed in TX-TL. Analysis of the product via UV-Vis absorption and liquid chromatography-mass spectrometry detected 4.95 mM of violacein. Expression levels of pathway enzymes were modeled using the TX-TL Toolbox. The model revealed the length of an enzyme coding sequence (CDS) significantly affected its expression level. Finally, pathway exploration suggested an improvement in violacein production at high VioC and VioD DNA concentrations.
Type Conference paper
URL http://biorxiv.org/content/early/2015/09/27/027656
Tag nwgm16-seed
ID 2016b
Funding GBMF PMTI
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