TY - JOUR T1 - Design Space Exploration of the Violacein Pathway in <em>Escherichia coli</em> Based Transcription Translation Cell-Free System (TX-TL) JF - bioRxiv DO - 10.1101/027656 SP - 027656 AU - Phuc H.B. Nguyen AU - Yong Y. Wu AU - Shaobin Guo AU - Richard M. Murray Y1 - 2015/01/01 UR - http://biorxiv.org/content/early/2015/09/27/027656.abstract N2 - 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’s coding sequence (CDS) significantly affected its expression level. Finally, pathway exploration suggested an improvement in violacein production at high VioC and VioD DNA concentrations. ER -