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.

中文翻译：

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三氨基三硝基苯（TATB）前体间苯三酚（PG）生物生产的生物工程。

生物工程需要选择合适的底盘，编辑其基因组，结合不同来源物种的性状，并通过合成电路控制基因。例如，当需要一个菌株来实现新的目标目标时，可以生产出所需的化学药品，那么最好的菌株，基因，技术，软件和专业知识就可以分布在各个实验室中。在这里，我们报告了一个项目，其中我们分配了间苯三酚（PG）作为目标，然后将整个美国陆军，海军和空军服务实验室的独特能力与设计产生这种分子的有机体的共同目标相结合。除实验室菌株大肠杆菌外，还从土壤和海水中筛选出了生物。推定的产生PG的酶是从飞机和油库中分离出的细菌菌株中提取的。最好的酶被引入海洋菌株大西洋海洋杆菌CP1中，其基因组经过编辑，可以重定向天然脂肪酸酯（FAE）生产中的碳通量。还尝试在枯草芽孢杆菌和丙酮丁醇梭菌中生产PG。在大肠杆菌中构建了一个遗传回路，对PG的积累作出反应，然后将其移植到体外纸基系统中，该系统可以用作未来低成本菌株筛选或现场感测的平台。这些努力共同表明，如何将具有特定领域专业知识的分布式生物技术实验室组织起来，以快速为目标生物工程项目提供解决方案，