(174ch) Biosensor Enabled Microbial Strain Improvement for Bioplastics Manufacturing

Microbial production of bio-based products from renewable feedstocks is a promising sustainable alternative to petroleum-based products. Synthetic biological engineering can produce superior metabolic designs that can be easily identified by genetic biosensors coupled to a fluorescence readout; and applied to strain improvement, enzyme and metabolic pathway evolution. Cis,cis muconic acid (CCM) and b-ketoadipate (BKAP) are potential platform chemicals for the manufacturing of bioplastics. The metabolically robust Pseudomonas putida engineered for overproduction of CCM, suffered from poor growth physiology. A CCM biosensor was developed and coupled to flow cytometry to screen a library of strains adapted for better growth. The improved strains exhibited superior growth and > 60% increase in CCM titers. The responsible mutations identified through resequencing analysis were further modulated and synergistically explored. In another instance a BKAP biosensor was developed to screen for BKAP producing P. putida strain adapted to grow on corn stover hydrolysate, that exhibited better growth and improved productivity.