The Development of Platform-Based Technologies for the Optimization of Sustainably Produced Chemicals

Genomatica has established an integrated computational/experimental bioengineering platform to design, create, and optimize novel high-producing organisms and bioprocesses. Here we present the use of our platform to develop E. coli strains for the production of the industrial chemical 1,4-butanediol (BDO) from carbohydrate feedstocks. These efforts have culminated in commercial scale production of over 5M lbs of BDO from dextrose. BDO is a chemical intermediate (>3B lbs/yr) produced commercially through energy-intensive petrochemical processes from hydrocarbon feedstocks and is used in a variety of products including automotive, electronics and apparel. Therefore, this product represents an opportunity to make a significant impact on the replacement of traditional petrochemical processes with benign bioprocesses using renewable feedstocks.

We are continuing to enhance our technology platform, enabling faster development of new processes with increased chance of success. Here we describe the application of cell-free transcription-translation (TX-TL), towards improving the design-build-test-learn cycle for the construction of a high-performing microorganism capable of producing BDO and other industrial chemicals from carbohydrates. We used systems biology approaches including ¹³C-flux analysis, proteomics, and metabolomics to characterize TX-TL applied to different engineered metabolic pathways, including BDO, and compared the results to the analogous in vivo processes. These studies provide critical insight into the active metabolism of TX-TL and mechanisms of redox and energy regeneration. Learnings from these studies were applied towards improving the TX-TL platform for the production of intermediate and specialty chemicals. The presentation will highlight the successful integration of the TX-TL technology into our rational and high-throughput strain engineering platform, and will show the potential advantages for using cell-free systems towards the optimization of sustainably produced chemicals.