Researchers from the Joint BioEnergy Institute have used advanced computing tools to engineer the bacteria Pseudomonas putida (P. putida) for isoprenol production as a precursor for sustainable aviation fuel. An open-access paper on their work is published in the journal Metabolic Engineering.

Isoprenol is a chemical involved in the production of a jet biofuel blendstock called 1,4-dimethylcyclooctane (DMCO). Researchers have produced isoprenol in several microbial hosts. However, efforts to make sustainable aviation fuel would benefit if isoprenol could be made in microorganisms that use fermentable sugars from plant material as a source of carbon. P. putida could be such a microorganism, but it needs engineering to be an optimal choice.

The team used computational modeling to predict targets for gene editing and to optimize metabolism in P. putida to maximize the production of isoprenol. This approach allowed the researchers to select and prioritize gene editing targets and therefore to test a smaller number of engineered strains.

Genome-scale metabolic and pathway engineering for production of the precursor of sustainable aviation fuel DMCO (1,4-dimethylcyclooctane), isoprenol, in Pseudomonas putida. Banerjee et al.

In addition, they applied known genetic edits to further improve isoprenol production and used proteomics to optimize the process.

The research achieved a 3.5 grams per liter isoprenol production titer, the highest reported for P. putida. The researchers concluded that their pathway optimization therefore resulted in a 10-fold improvement of isoprenol in P. putida. The researchers suggest additional improvements must be made to improve isoprenol yields for industrial applications.

Commercial-scale production of isoprenol and DMCO at commercial scale still requires additional improvements such as the inclusion of CRISPR gene editing and other bioprocess technologies.

The research was funded by the Joint BioEnergy Institute supported by the Department of Energy Office of Science, Biological and Environmental Research Program.

Resources

• Deepanwita Banerjee, Ian S. Yunus, Xi Wang, Jinho Kim, Aparajitha Srinivasan, Russel Menchavez, Yan Chen, Jennifer W. Gin, Christopher J. Petzold, Hector Garcia Martin, Jon K. Magnuson, Paul D. Adams, Blake A. Simmons, Aindrila Mukhopadhyay, Joonhoon Kim, Taek Soon Lee (2024) “Genome-scale and pathway engineering for the sustainable aviation fuel precursor isoprenol production in Pseudomonas putida,” Metabolic Engineering, Volume 82, Pages 157-170 doi: 10.1016/j.ymben.2024.02.004