9-04: The design, engineering, and optimization of a microbial process for manufacturing of 1,4-butanediol

Genomatica has utilized its computational and experimental competencies in an integrated metabolic engineering platform to design, create, and optimize novel organisms and bioprocesses.  We present the use of our platform to develop E. coli strains for the production of the industrial chemical 1,4-butanediol (BDO) from glucose and sucrose. BDO is a four-carbon diol that currently is manufactured exclusively through various petrochemical routes. It is part of a large volume family of solvents and polymer intermediates with an overall market opportunity exceeding $4.0B. Therefore, this is an opportunity to make a significant impact on the replacement of traditional petrochemical processes with relatively benign bioprocesses using renewable feedstocks.

Here we describe using this technology platform to design and construct a high-performing microorganism capable of producing BDO from carbohydrates. The Biopathway Predictor algorithm was employed to reveal all possible routes to BDO from central metabolites, and choose the most favorable for implementation. We then utilized the OptKnock methodology to identify a set of gene deletions designed to couple BDO formation to growth. After design-based production of the biocatalyst, our models facilitated the analysis of fermentation data to evaluate performance. Systems biology approaches including microarrays, 13C-flux analysis, and metabolomics were applied to characterize the strain, identify targets for further improvement, and optimize the fermentation process. The presentation will show our progress in BDO titer, production rate, and yield through various strain and process improvements, ultimately resulting in an economically attractive process that is being validated at the pilot and demo scale.