(86e) Development of a Genetic Toolset for Acinetobacter Baylyi (ADP1), a Host for Lignin-Based Metabolic Engineering

Efficient valorization of carbon sources currently treated as waste represents a key growth sector for metabolic engineering, especially as the field looks to advance sustainability efforts. Environmental microbes possess remarkable capacity to assimilate and repurpose a wide variety of feedstocks, and leveraging this ability provides opportunity for both remediation and synthesis of value-added compounds. One interesting material to this end is lignin, the most abundant aromatic polymer in nature, which is currently treated as a waste product. Several organisms have shown capacity to degrade and consume lignin, but the the facile genetics of the bacterium Acinetobacter baylyi (ADP1) make it an exciting potential model organism for lignin bioprocessing. However, this host currently lacks the necessary synthetic biology tools for desired applications. Here, we establish easy and rapid molecular cloning workflows, a promoter library, RBS variants, and an unprecedented number of bacterial chromosomal expression tools to enable a wide variety of synthetic biology applications in ADP1. In addition, we demonstrate the efficacy of these tools for lignin-based metabolic engineering.