Evolution of A. baylyi to grow using a PCA meta-cleavage pathway

The native route for aromatic compound catabolism in Acinetobacter baylyi ADP1 was replaced with a foreign pathway that uses different enzymes, metabolites, and redox carriers. The goal was to improve biotechnology applications involving aromatic compound biotransformation. One chromosomal copy of the nonnative genes for protocatechuate (PCA) degradation via a 2,3-cleavage route failed to enable growth on the target substrates. To enable growth, genomic segments encompassing the foreign genes were amplified to establish tandem head-to-tail arrays of heterologous pra-genes. This gene dosage change enabled growth on 4 hydroxybenzoate (4HB) or PCA as the sole carbon source. During adaptive laboratory evolution, compensatory mutations arose that obviated the need for multiple gene copies. Whole genome sequence data were generated for strains isolated before, during, and after adaptive laboratory evolution.