title: Proteomic analysis of organic sulfur compound utilisation in Advenella mimigardefordensis strain DPN7T.
abstract: 2-Mercaptosuccinate (MS) and 3,3´-ditiodipropionate (DTDP) were discussed as precursor substance for production of polythioesters (PTE). Therefore, degradation of MS and DTDP was investigated in Advenella mimigardefordensis strain DPN7T, applying differential proteomic analysis, gene deletion and enzyme assays. Protein extracts of cells cultivated with MS, DTDP or 3-sulfinopropionic acid (SP) were compared with those cultivated with propionate (P) and/or succinate (S). The chaperone DnaK (ratio DTDP/P 9.2, 3SP/P 4.0, MS/S 6.1, DTDP/S 6.2) and a Do-like serine protease (DegP) were increased during utilization of all organic sulfur compounds. Furthermore, a putative bacterioferritin (locus tag MIM_c12960) showed high abundance (ratio DTDP/P 5.3, 3SP/P 3.2, MS/S 4.8, DTDP/S 3.9) and is probably involved in a thiol-specific stress response. The deletion of two genes encoding transcriptional regulators (LysR (MIM_c31370) and Xre (MIM_c31360)) in the close proximity of the relevant genes of DTDP catabolism (acdA, mdo and the genes encoding the enzymes of the methylcitric acid cycle; prpC,acnD, prpF and prpB) showed that these two regulators are essential for growth of A. mimigardefordensis strain DPN7T with DTDP and that they most probably regulate transcription of genes mandatory for this catabolic pathway. Furthermore, proteome analysis revealed a high abundance (ratio MS/S 10.9) of a hypothetical cupin-2-domain containing protein (MIM_c37420). This protein shows an amino acid sequence similarity of 60% to a newly identified MS dioxygenase from Variovorax paradoxus strain B4. Deletion of the gene and the adjacently located transcriptional regulator LysR, as well as heterologous expression of MIM_c37420, the putative mercaptosuccinate dioxygenase (Msdo) from A. mimigardefordensis, showed that this protein is the key enzyme of MS degradation in A. mimigardefordensis strain DPN7T (KM 0.2 mM, specific activity 17.1 μmol mg-1 min-1) and is controlled by LysR (MIM_c37410).
authors.affiliation: Institut für Molekulare Mikrobiologie und Biotechnologie, Westfälische Wilhelms-Universität, Münster, Germany.
authors.forename: Christina
authors.initials: C
authors.lastname: Meinert
authors.affiliation: Institut für Molekulare Mikrobiologie und Biotechnologie, Westfälische Wilhelms-Universität, Münster, Germany.
authors.forename: Ulrike
authors.initials: U
authors.lastname: Brandt
authors.affiliation: Institut für Molekulare Mikrobiologie und Biotechnologie, Westfälische Wilhelms-Universität, Münster, Germany.
authors.forename: Viktoria
authors.initials: V
authors.lastname: Heine
authors.affiliation: Institut für Molekulare Mikrobiologie und Biotechnologie, Westfälische Wilhelms-Universität, Münster, Germany.
authors.forename: Jessica
authors.initials: J
authors.lastname: Beyert
authors.affiliation: Institut für Molekulare Mikrobiologie und Biotechnologie, Westfälische Wilhelms-Universität, Münster, Germany.
authors.forename: Sina
authors.initials: S
authors.lastname: Schmidl
authors.affiliation: Institut für Molekulare Mikrobiologie und Biotechnologie, Westfälische Wilhelms-Universität, Münster, Germany.
authors.forename: Jan Hendrik
authors.initials: JH
authors.lastname: Wübbeler
authors.affiliation: Institut für Mikrobiologie, Ernst-Moritz-Arndt-Universität, Greifswald, Germany.
authors.forename: Birgit
authors.initials: B
authors.lastname: Voigt
authors.affiliation: Institut für Mikrobiologie, Ernst-Moritz-Arndt-Universität, Greifswald, Germany.
authors.forename: Katharina
authors.initials: K
authors.lastname: Riedel
authors.affiliation: Institut für Molekulare Mikrobiologie und Biotechnologie, Westfälische Wilhelms-Universität, Münster, Germany.|Environmental Science Department, King Abdulaziz University, Jeddah, Saudi Arabia.
authors.forename: Alexander
authors.initials: A
authors.lastname: Steinbüchel
mesh_terms: D042401:Alcaligenaceae / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D013058:Mass Spectrometry; D009930:Organic Chemicals / Q000378:metabolism*; D011422:Propionates / Q000378:metabolism; D020543:Proteome / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D012984:Software; D013870:Thiomalates / Q000378:metabolism
chemical_list: D009930:Organic Chemicals; D011422:Propionates; D020543:Proteome; D013870:Thiomalates; C009351:3-sulfinopropionic acid; C046062:2-thiomalic acid