Development of SynBio tools to more predictably clone, express and select biocatalytic activities for metabolic pathway optimization

The deployment of bio-based manufacturing requires the optimization of multicomponent biosynthetic pathways to generate intermediates and commercial end products.  However, unlike physical/computer engineering, the rules for predictive bioengineering are not well understood.  I will describe the application of synthetic biology tools to enable predictably engineer biological production systems with reference to commercially relevant examples.  These tools include protein engineering to address poor response controls for gene expression, the development of synthetic landing pads to optimize the genomic operating environment around delivered genes, the use of genome editing and RNA trafficking systems to improve gene expression, the application of transciptomics and metabolomics to enhance cell system performance, the development of synthetic regulatory elements to better control gene expression and the deployment of our proprietary inABLE combinatorial genetics platform for large scale gene/pathway assembly.  Together these have been used to rapidly clone, express, select and optimize target activities for many enzymatic reactions, from thousands of independent genes derived from metagenomic and phylogenetic discovery.  Obvious synergy exists between this approach and versatile, solid phase screening and selection methods using growth-based, crossfeeding or colorimetric methods to identify engineered cells of interest.  This is illustrated through the rapid identification of critical pathway enzymes, optimal gene coding sequences and enzyme variants from inABLE^®-derived high quality variant libraries for industrial applications in bio-based polymers, chemicals and personal care products.  In developing this suite of technologies we aims to bring increasing predictability and overcome persistent limitations associated with today’s iterative and empirical processes for microbial strain improvement.