RT Journal Article
SR Electronic
T1 Design Space Exploration of the Violacein Pathway in <em>Escherichia coli</em> Based Transcription Translation Cell-Free System (TX-TL)
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 027656
DO 10.1101/027656
A1 Phuc H.B. Nguyen
A1 Yong Y. Wu
A1 Shaobin Guo
A1 Richard M. Murray
YR 2015
UL http://biorxiv.org/content/early/2015/09/27/027656.abstract
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. 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.