title: Regulation of solvent tolerance in Pseudomonas putida S12 mediated by mobile elements.
abstract: Organic solvent-tolerant bacteria are outstanding and versatile hosts for the bio-based production of a broad range of generally toxic aromatic compounds. The energetically costly solvent tolerance mechanisms are subject to multiple levels of regulation, involving among other mobile genetic elements. The genome of the solvent-tolerant Pseudomonas putida S12 contains many such mobile elements that play a major role in the regulation and adaptation to various stress conditions, including the regulation of expression of the solvent efflux pump SrpABC. We recently sequenced the genome of P. putida S12. Detailed annotation identified a threefold higher copy number of the mobile element ISS12 in contrast to earlier observations. In this study, we describe the mobile genetic elements and elaborate on the role of ISS12 in the establishment and maintenance of solvent tolerance in P. putida. We identified three different variants of ISS12 of which a single variant exhibits a high translocation rate. One copy of this variant caused a loss of solvent tolerance in the sequenced strain by disruption of srpA. Solvent tolerance could be restored by applying selective pressure, leading to a clean excision of the mobile element.
authors.affiliation: Microbial Biotechnology and Health, Institute of Biology, Leiden University, Leiden, The Netherlands.
authors.forename: Rohola
authors.initials: R
authors.lastname: Hosseini
authors.affiliation: Institute of Applied Microbiology - iAMB, Aachen Biology and Biotechnology - ABBt, RWTH Aachen University, Aachen, Germany.
authors.forename: Jannis
authors.initials: J
authors.lastname: Kuepper
authors.affiliation: Institute of Applied Microbiology - iAMB, Aachen Biology and Biotechnology - ABBt, RWTH Aachen University, Aachen, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Koebbing
authors.affiliation: Institute of Applied Microbiology - iAMB, Aachen Biology and Biotechnology - ABBt, RWTH Aachen University, Aachen, Germany.
authors.forename: Lars M
authors.initials: LM
authors.lastname: Blank
authors.affiliation: Institute of Applied Microbiology - iAMB, Aachen Biology and Biotechnology - ABBt, RWTH Aachen University, Aachen, Germany.
authors.forename: Nick
authors.initials: N
authors.lastname: Wierckx
authors.affiliation: Microbial Biotechnology and Health, Institute of Biology, Leiden University, Leiden, The Netherlands.
authors.forename: Johannes H
authors.initials: JH
authors.lastname: de Winde
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D020071:Interspersed Repetitive Sequences; D026901:Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism; D016254:Mutagenesis, Insertional; D016958:Pseudomonas putida / Q000187:drug effects* / Q000235:genetics* / Q000378:metabolism; D012997:Solvents / Q000494:pharmacology*
chemical_list: D001426:Bacterial Proteins; D026901:Membrane Transport Proteins; D012997:Solvents