Organism Engineering for the Bioproduction of the Triaminotrinitrobenzene (TATB) Precursor Phloroglucinol (PG)

Adam Meyer, Ishtiaq Saaem, Adam Silverman, Vanessa A. Varaljay, Rebecca Mickol, Steven Blum, Alexander V. Tobias, Nathan D. Schwalm, Wais Mojadedi, Elizabeth Onderko, Cassandra Bristol, Shangtao Liu, Katelin Pratt, Arturo Casini, Raissa Eluere, Felix Moser, Carrie Drake, Maneesh Gupta, Nancy Kelley-Loughnane, Julius P. LucksKatherine L. Akingbade, Matthew P. Lux, Sarah Glaven, Wendy Crookes-Goodson, Michael C. Jewett, D. Benjamin Gordon, Christopher A. Voigt*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

Abstract

Organism engineering requires the selection of an appropriate chassis, editing its genome, combining traits from different source species, and controlling genes with synthetic circuits. When a strain is needed for a new target objective, for example, to produce a chemical-of-need, the best strains, genes, techniques, software, and expertise may be distributed across laboratories. Here, we report a project where we were assigned phloroglucinol (PG) as a target, and then combined unique capabilities across the United States Army, Navy, and Air Force service laboratories with the shared goal of designing an organism to produce this molecule. In addition to the laboratory strain Escherichia coli, organisms were screened from soil and seawater. Putative PG-producing enzymes were mined from a strain bank of bacteria isolated from aircraft and fuel depots. The best enzyme was introduced into the ocean strain Marinobacter atlanticus CP1 with its genome edited to redirect carbon flux from natural fatty acid ester (FAE) production. PG production was also attempted in Bacillus subtilis and Clostridium acetobutylicum. A genetic circuit was constructed in E. coli that responds to PG accumulation, which was then ported to an in vitro paper-based system that could serve as a platform for future low-cost strain screening or for in-field sensing. Collectively, these efforts show how distributed biotechnology laboratories with domain-specific expertise can be marshalled to quickly provide a solution for a targeted organism engineering project, and highlights data and material sharing protocols needed to accelerate future efforts.

Original languageEnglish (US)
Pages (from-to)2746-2755
Number of pages10
JournalACS synthetic biology
Volume8
Issue number12
DOIs
StatePublished - Dec 20 2019

Keywords

  • TX-TL
  • Tri-Service
  • cell-free sensing
  • enzyme mining
  • metabolic engineering
  • military environments
  • synthetic biology

ASJC Scopus subject areas

  • Biomedical Engineering
  • Biochemistry, Genetics and Molecular Biology (miscellaneous)

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