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A novel toxin-antitoxin module SlvT-SlvA governs megaplasmid stability and incites solvent tolerance in Pseudomonas putida S12

By Hadiastri Kusumawardhani, David van Dijk, Rohola Hosseini, Johannes H. de Winde

Posted 03 Jan 2020
bioRxiv DOI: 10.1101/2020.01.02.893495

Pseudomonas putida S12 is highly tolerant towards organic solvents in saturating concentrations, rendering this microorganism suitable for the industrial production of various aromatic compounds. Previous studies reveal that P. putida S12 contains a single-copy 583 kbp megaplasmid pTTS12. This pTTS12 encodes several important operons and gene clusters facilitating P. putida S12 to survive and grow in the presence of toxic compounds or other environmental stresses. We wished to revisit and further scrutinize the role of pTTS12 in conferring solvent tolerance. To this end, we cured the megaplasmid from P. putida S12 and conclusively confirmed that the SrpABC efflux pump is the major contributor of solvent tolerance on the megaplasmid pTTS12. Importantly, we identified a novel toxin-antitoxin module (proposed gene names slvT and slvA respectively) encoded on pTTS12 which contributes to the solvent tolerant phenotype and is essential in conferring genetic stability to the megaplasmid. Chromosomal introduction of the srp operon in combination with slvAT gene pair created a solvent tolerance phenotype in non-solvent tolerant strains such as P. putida KT2440, E. coli TG1, and E. coli BL21(DE3).

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