TY - JOUR
T1 - Organism Engineering for the Bioproduction of the Triaminotrinitrobenzene (TATB) Precursor Phloroglucinol (PG)
AU - Meyer, Adam
AU - Saaem, Ishtiaq
AU - Silverman, Adam
AU - Varaljay, Vanessa A.
AU - Mickol, Rebecca
AU - Blum, Steven
AU - Tobias, Alexander V.
AU - Schwalm, Nathan D.
AU - Mojadedi, Wais
AU - Onderko, Elizabeth
AU - Bristol, Cassandra
AU - Liu, Shangtao
AU - Pratt, Katelin
AU - Casini, Arturo
AU - Eluere, Raissa
AU - Moser, Felix
AU - Drake, Carrie
AU - Gupta, Maneesh
AU - Kelley-Loughnane, Nancy
AU - Lucks, Julius P.
AU - Akingbade, Katherine L.
AU - Lux, Matthew P.
AU - Glaven, Sarah
AU - Crookes-Goodson, Wendy
AU - Jewett, Michael C.
AU - Gordon, D. Benjamin
AU - Voigt, Christopher A.
N1 - Funding Information:
The authors wish to acknowledge the Biological Materials and Processes Research Team, Materials and Manufacturing Directorate AFRL for the aircraft strain bank and Dr. Chia Hung for helpful discussions on project direction and experimental designs. This work was funded by the U.S. Defense Advanced Research Projects Agency (DARPA) awards HR0011-15-0084 and FA8750-17-C-0229, the Office of Naval Research Multidisciplinary University Research Initiative Award N00014-16-1-2388 the Army Research Office Multidisciplinary University Research Initiative Awards W911NF-18-1-0200 and W911NF-16-1-0372, the Air Force Research Laboratory Center of Excellence Grant FA8650-15-2-5518, the Institute for Collaborative Biotechnologies through awards W911NF-09-0001 and W911NF-19-2-0026 with the U.S. Army Research Office, the Office of Naval Research Vannevar Bush Faculty Fellowship (formerly NSSEFF) N00014-16-1-2509, the Laboratory University Collaboration Initiative (LUCI) program, and the Synthetic Biology for Military Environments Applied Research for the Advancement of S&T Priorities (ARAP) program of the U.S. Office of the Under Secretary of Defense for Research and Engineering. The research reported in this publication has been cleared for public release under reference number 88ABW-2019-4584.
Funding Information:
The authors wish to acknowledge the Biological Materials and Processes Research Team, Materials and Manufacturing Directorate, AFRL for the aircraft strain bank and Dr. Chia Hung for helpful discussions on project direction and experimental designs. This work was funded by the U.S. Defense Advanced Research Projects Agency (DARPA) awards HR0011-15-0084 and FA8750-17-C-0229, the Office of Naval Research Multidisciplinary University Research Initiative Award N00014-16-1-2388, the Army Research Office Multidisciplinary University Research Initiative Awards W911NF-18-1-0200 and W911NF-16-1-0372, the Air Force Research Laboratory Center of Excellence Grant FA8650-15-2-5518, the Institute for Collaborative Biotechnologies through awards W911NF-09-0001 and W911NF-19-2-0026 with the U.S. Army Research Office, the Office of Naval Research Vannevar Bush Faculty Fellowship (formerly NSSEFF) N00014-16-1-2509, the Laboratory University Collaboration Initiative (LUCI) program, and the Synthetic Biology for Military Environments Applied Research for the Advancement of S&T Priorities (ARAP) program of the U.S. Office of the Under Secretary of Defense for Research and Engineering. The research reported in this publication has been cleared for public release under reference number 88ABW-2019-4584.
Publisher Copyright:
Copyright © 2019 American Chemical Society.
PY - 2019/12/20
Y1 - 2019/12/20
N2 - 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.
AB - 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.
KW - TX-TL
KW - Tri-Service
KW - cell-free sensing
KW - enzyme mining
KW - metabolic engineering
KW - military environments
KW - synthetic biology
UR - http://www.scopus.com/inward/record.url?scp=85076755003&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85076755003&partnerID=8YFLogxK
U2 - 10.1021/acssynbio.9b00393
DO - 10.1021/acssynbio.9b00393
M3 - Article
C2 - 31750651
AN - SCOPUS:85076755003
VL - 8
SP - 2746
EP - 2755
JO - ACS Synthetic Biology
JF - ACS Synthetic Biology
SN - 2161-5063
IS - 12
ER -