A novel reporter system for bacterial and mammalian cells based on the non-ribosomal peptide indigoidine.
Metab Eng
; 14(4): 325-35, 2012 Jul.
Article
em En
| MEDLINE
| ID: mdl-22543310
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.
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1
Base de dados:
MEDLINE
Assunto principal:
Piperidonas
/
Streptomyces
Idioma:
En
Ano de publicação:
2012
Tipo de documento:
Article