Back to Search Start Over

A novel reporter system for bacterial and mammalian cells based on the non-ribosomal peptide indigoidine

Authors :
Martin Fussenegger
David Ausländer
Christian Kemmer
Simon Ausländer
Marius Müller
Source :
Metabolic engineering
Publication Year :
2012

Abstract

The biosynthesis of non-ribosomal peptides, many of which have pharmaceutical activities, is an evolutionary privilege of microorganisms. Capitalizing on the universal set of the Streptomyces lavendulae non-ribosomal peptide synthase BpsA and the Streptomyces verticillus 4'-phosphopantetheinyl transferase Svp, we have engineered Escherichia coli as well as mammalian cells, including human stem cells, to produce the blue 3,3'-bipyridyl pigment keto-indigoidine and the reduced colorless but fluorescent leuco-isoform. Detailed characterization of a tailored substrate-free chromogenic assay and FACS analysis showed that indigoidine's blue color and fluorescence could be reliably profiled in bacteria and mammalian cells using standard multiwell-compatible detection equipment. Besides serving as an inexpensive, reliable, versatile and easy-to-assay cross-kingdom reporter system, the potential of having mammalian cells produce non-ribosomal peptides, preferably ones with biopharmaceutical activities, may provide novel treatment opportunities in future gene- and cell-based therapies.

Details

Volume :
14
Issue :
4
Database :
OpenAIRE
Journal :
Metabolic engineering
Accession number :
edsair.doi.dedup.....77b93d68b8d63ea8bd82d6ff5c4294b8
Full Text :
https://doi.org/10.1016/j.ymben.2012.04.002