Rapid Development of Acetogenic Clostridia using Highly Multiplexed Genome Engineering for Control of C1 Bioconversion

This project will develop a commercially scalable emerging model organism, called Clostridium autoethanogenum, that converts a single carbon (C1) feedstock (carbon dioxide, carbon monoxide, etc. from waste gas emissions) into 3-hydroxyproprionic acid. 3-hydroxypropionic acid is an ideal biorenewable precursor to industrially important polymers such as acrylates. To do this, we will apply several systems and synthetic biology technologies, coupling together algorithmic design approaches, highly multiplexed genome-scale engineering techniques, and omics measurements, to exert complete control over the metabolism of Clostridium autoethanogenum. First, we will employ an integrated computational-experimental approach to engineer optimized biosynthesis pathways for 3-hydroxypropionic acid in Clostridium autoethanogenum. Second, to redirect metabolic flows towards 3- hydroxypropionic acid production, we will develop and demonstrate a very highly multiplexed version of CRISPR that utilizes highly non-repetitive genetic parts to up-regulate or down-regulate up to 20 targeted genes simultaneously. Third, we will perform technoeconomic assessments of C1 bioconversion to 3-hydroxypropionic acid and couple those assessments to algorithm-designed genetic modifications, determining genotype-phenotype-cost relationships across several metrics. This project will result in a commercially scalable emerging model organism capable of producing 3-hydroxypropionic acid at economically competitive, high productivities from low-cost C1 feedstock. This project is a collaboration with LanzaTech Inc.