Novel Toxin-Antitoxin Module SlvT-SlvA Regulates Megaplasmid Stability and Incites Solvent Tolerance in Pseudomonas putida S12

Applied and Environmental Microbiology, vol. 86

Abstract

Sustainable alternatives for high-value chemicals can be achieved by using renewable feedstocks in bacterial biocatalysis. However, during the bioproduction of such chemicals and biopolymers, aromatic compounds that function as products, substrates, or intermediates in the production process may exert toxicity to microbial host cells and limit the production yield. Therefore, solvent tolerance is a highly preferable trait for microbial hosts in the biobased production of aromatic chemicals and biopolymers. In this study, we revisit the essential role of megaplasmid pTTS12 from solvent-tolerant Pseudomonas putida S12 for molecular adaptation to an organic solvent. In addition to the solvent extrusion pump (SrpABC), we identified a novel toxin-antitoxin module (SlvAT) which contributes to short-term tolerance in moderate solvent concentrations, as well as to the stability of pTTS12. These two gene clusters were successfully expressed in non-solvent-tolerant strains of P. putida and Escherichia coli strains to confer and enhance solvent tolerance.

Authors 4

1. Hadiastri Kusumawardhani Aachen Institute of Biology Leiden

   Leiden University

   Affiliation as printed

   Institute of Biology Leiden, Leiden University, Leiden, The Netherlands

2. David van Dijk Aachen Institute of Biology Leiden

   Leiden University

   Affiliation as printed

   Institute of Biology Leiden, Leiden University, Leiden, The Netherlands

3. Rohola Hosseini Aachen Institute of Biology Leiden

   Leiden University

   Affiliation as printed

   Institute of Biology Leiden, Leiden University, Leiden, The Netherlands

4. Johannes H. DE WINDE corresponding Aachen Institute of Biology Leiden

   Leiden University

   Affiliation as printed

   Institute of Biology Leiden, Leiden University, Leiden, The Netherlands