Generation of <i>Pseudomonas putida</i> KT2440 Strains with Efficient Utilization of Xylose and Galactose via Adaptive Laboratory Evolution

While Pseudomonas putida KT2440 has great potential for biomass-converting processes, its inability to utilize the biomass abundant sugars xylose and galactose limited applications. In this study, we utilized Adaptive Laboratory Evolution (ALE) optimize engineered with heterologous expression of xylD encoding xylonate dehydratase from Caulobacter crescentus galETKM UDP-glucose 4-epimerase, galactose-1-phosphate uridylyltransferase, galactokinase, galactose-1-epimerase Escherichia coli K-12 MG1655. Poor starting strain growth (<0.1 h–1 or none) was evolutionarily optimized rates up 0.25 on 0.52 galactose. Whole-genome sequencing, transcriptomic analysis, screens revealed significant roles kguT a 2-ketogluconate operon repressor transporter, gtsABCD an ATP-binding cassette (ABC) sugar transporting system in conditions, respectively. Finally, expressed indigoidine production pathway evolved unevolved strains successfully produced 3.2 g/L 2.2 10 either whereas did not produce any detectable product. Thus, generated have broad application as platform chassis develop efficient microorganism-based biomass-utilizing bioprocesses.