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

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.

journal_name

Metab Eng

journal_title

Metabolic engineering

authors

Müller M,Ausländer S,Ausländer D,Kemmer C,Fussenegger M

doi

10.1016/j.ymben.2012.04.002

subject

Has Abstract

pub_date

2012-07-01 00:00:00

pages

325-35

issue

4

eissn

1096-7176

issn

1096-7184

pii

S1096-7176(12)00045-6

journal_volume

14

pub_type

杂志文章

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