title: Aerobic degradation of mercaptosuccinate by the gram-negative bacterium Variovorax paradoxus strain B4.
abstract: The Gram-negative bacterium Variovorax paradoxus strain B4 was isolated from soil under mesophilic and aerobic conditions to elucidate the so far unknown catabolism of mercaptosuccinate (MS). During growth with MS this strain released significant amounts of sulfate into the medium. Tn5::mob-induced mutagenesis was successfully employed and yielded nine independent mutants incapable of using MS as a carbon source. In six of these mutants, Tn5::mob insertions were mapped in a putative gene encoding a molybdenum (Mo) cofactor biosynthesis protein (moeA). In two further mutants the Tn5::mob insertion was mapped in the gene coding for a putative molybdopterin (MPT) oxidoreductase. In contrast to the wild type, these eight mutants also showed no growth on taurine. In another mutant a gene putatively encoding a 3-hydroxyacyl-coenzyme A dehydrogenase (paaH2) was disrupted by transposon insertion. Upon subcellular fractionation of wild-type cells cultivated with MS as sole carbon and sulfur source, MPT oxidoreductase activity was detected in only the cytoplasmic fraction. Cells grown with succinate, taurine, or gluconate as a sole carbon source exhibited no activity or much lower activity. MPT oxidoreductase activity in the cytoplasmic fraction of the Tn5::mob-induced mutant Icr6 was 3-fold lower in comparison to the wild type. Therefore, a new pathway for MS catabolism in V. paradoxus strain B4 is proposed: (i) MPT oxidoreductase catalyzes the conversion of MS first into sulfinosuccinate (a putative organo-sulfur compound composed of succinate and a sulfino group) and then into sulfosuccinate by successive transfer of oxygen atoms, (ii) sulfosuccinate is cleaved into oxaloacetate and sulfite, and (iii) sulfite is oxidized to sulfate.
authors.affiliation: Institut für Molekulare Mikrobiologie und Biotechnologie, Westfälische Wilhelms-Universität Münster, D-48149 Münster, Germany.
authors.forename: Irma
authors.initials: I
authors.lastname: Carbajal-Rodríguez
authors.forename: Nadine
authors.initials: N
authors.lastname: Stöveken
authors.forename: Barbara
authors.initials: B
authors.lastname: Satola
authors.forename: Jan Hendrik
authors.initials: JH
authors.lastname: Wübbeler
authors.forename: Alexander
authors.initials: A
authors.lastname: Steinbüchel
mesh_terms: D015094:3-Hydroxyacyl CoA Dehydrogenases / Q000235:genetics / Q000378:metabolism; D000332:Aerobiosis; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D016000:Cluster Analysis; D042621:Comamonadaceae / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D003593:Cytoplasm / Q000201:enzymology; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D005942:Gluconates / Q000378:metabolism; D053858:Metabolic Networks and Pathways / Q000235:genetics; D008954:Models, Biological; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D010802:Phylogeny; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D017422:Sequence Analysis, DNA; D012988:Soil Microbiology; D013431:Sulfates / Q000378:metabolism; D013466:Sulfurtransferases / Q000235:genetics / Q000378:metabolism; D013654:Taurine / Q000378:metabolism; D013696:Temperature; D013870:Thiomalates / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D005942:Gluconates; D012336:RNA, Ribosomal, 16S; D013431:Sulfates; D013870:Thiomalates; D013654:Taurine; C046062:2-thiomalic acid; D015094:3-Hydroxyacyl CoA Dehydrogenases; D013466:Sulfurtransferases; C030691:gluconic acid