Innovative, Scalable and Sustainable Bio-manufacturing Routes to Precursors of the High Explosive Hexanitrohexaazaisowurtzitane (CL-20)

Nigel Scrutton | University of Manchester



To develop innovative, scalable and environmentally sustainable bio-manufacturing routes to benzylamine. This will enable the development of a more cost effective and semi-synthetic route to the high explosive hexanitrohexaazaisowurtzitane (CL-20). This project will assist the Department of Defense (DoD) by generating an alternative strategy that will reduce the overall costs and environmental challenges associated with the production of CL-20.

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Technical Approach

We will construct stable, biosynthetic benzylamine-producing deoxyribonucleic acid (DNA) constructs initially within Escherichia coli (E. coli) and then within Halomonas, an existing robust bio-manufacturing host. The Halomonas chassis is key to maximize cost-effectiveness of the process, as industrial biosyntheses can be performed using seawater under non-sterile conditions in simplified bioreactors. This construct will be optimized to maximize titres using our proven automated metabolic engineering and synthetic biology strategies that utilize DESIGN, BUILD, TEST, LEARN platforms to develop the most highly efficient in vivo pathways. This refactoring strategy will establish the optimal genetic controlling elements (e.g. promoters) by identifying pathway bottlenecks (precursor accumulation) and designing the most effective route to overcome them. The high throughput pipeline will rapidly build and quantify the titres of the new microbial strains, enabling only the most efficient constructs to be transitioned into Halomonas. Our proprietary genetic tools for stable high copy construct maintenance combined with established chromosomal integration techniques will ensure stable strain development with high benzylamine titres. Optimization of fermentation strategies and downstream processing will enable efficient production, extraction and purification of benzylamine. Our collaboration with the Naval Air Warfare Center Weapons Division at China Lake will ascertain the successful utilization of bio-benzylamine into CL-20 manufacture.

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There is an urgent need to develop a more cost effective and sustainable route to the commodity chemical benzylamine, which is scalable and eliminates the need for highly toxic reagents, solvents and waste products. This will contribute enormously to reducing the overall costs and environmental challenges associated with the production of CL-20. A synthetic biology route towards benzylamine biosynthesis using the robust industrial strain Halomonas will satisfy these criteria, by eliminating toxic reagents and reducing manufacturing costs by using waste, renewable carbon sources and non-sanitized seawater or wastewater. The dramatic reduction in both the capital expenditure (CapEx) and operational expenditure (OpEx) costs of traditional bioreactors (up to 80%) will assist in strengthening the role of bio-manufacturing as a commercially viable and sustainable options for chemicals manufacture.

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Points of Contact

Principal Investigator

Dr. Nigel Scrutton

University of Manchester

Phone: 00441613065184