Whole-genome sequencing of solvent-tolerant Pseudomonas putida S12 (ATCC 700801)

This project focuses mainly on the mechanisms of Pseudomonas putida S12 solvent tolerance and its genome stability. P. putida S12 ATCC 700801 is widely tolerant towards various organic solvents. This strain has been used to produce aromatic chemicals and biotransformations . Recently, P. putida S12 genome have also been fully sequenced. The genome of P. putida S12 consists of 5.8-mega base pairs of chromosome Acc. no. CP009974 and a single copy of 580-kilo base pairs mega plasmid Acc. no. CP009975. An RND efflux pump SrpABC and styrene degradation pathway are encoded in the mega plasmid. These two systems play an important role in inciting solvent-tolerance trait of P. putida S12.In this study, we resequenced P. putida S12 obtained from ATCC repository. Megaplasmid was cured from this strain using mitomycin C and the resulting strains were sequenced to investigate the removal of megaplasmid pTTS12. This megaplasmid cured strains were adapted to higher toluene concentration and thus regained its solvent tolerance. SNPs and other mutations occurred in the adapted strains were mapped in this sequencing project.