PT  - JOURNAL ARTICLE
AU  - Phuc H.B. Nguyen
AU  - Yong Y. Wu
AU  - Shaobin Guo
AU  - Richard M. Murray
TI  - Design Space Exploration of the Violacein Pathway in <em>Escherichia coli</em> Based Cell-Free System
AID  - 10.1101/027656
DP  - 2016 Jan 01
TA  - bioRxiv
PG  - 027656
4099  - http://biorxiv.org/content/early/2016/01/26/027656.short
4100  - http://biorxiv.org/content/early/2016/01/26/027656.full
AB  - 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. The pathway was successfully reconstructed in TX-TL. Its variation produced different metabolites as evident from the extracts’ assorted colors. Analysis of the violacein product via UV-Vis absorption and liquid chromatography-mass spectrometry (LC-MS) detected 68 ng of violacein per μl reaction volume. Significant buildup of prodeoxyviolacein intermediate was also detected in the equimolar TX-TL reaction. Finally, design space exploration experiments suggested an improvement in violacein production at high VioC and VioD DNA concentrations.