(65e) Biorenewable Chemical Production from Pyrolysate By Lactic Acid Bacteria

Lignocellulosic biomass is an abundant source of carbon, especially in agricultural areas like Iowa. Fast pyrolysis is one method that can be used to create fermentable sugars from cellulosic sources, but product of this process, bio-oil, is rich in (1) levoglucosan, an anhydrosugar form of glucose, and (2) phenolic and other inhibitory compounds. Here we report the engineering of Lactococcus lactis to utilize purified levoglucosan for the production of enantiomerically pure L-lactic acid and, by expressing a heterologous three-gene pathway, 1,2-propanediol. We also report media conditions that allow the production of these chemicals from diluted bio-oil and strategies to engineer tolerance to bio-oil and other "dirty" substrates. Finally, we report the development of CRISPR-assisted genome-scale engineering and library creation tools in this important industrial organism.