Assimilation of homotaurine-nitrogen by<i>Burkholderia</i>sp. and excretion of sulfopropanoate

Mayer, Jutta; Denger, Karin; Kaspar, Katrin; Hollemeyer, Klaus; Smits, Theo H. M.; Huhn, Thomas; Cook, Alasdair M.

doi:10.1111/j.1574-6968.2007.01014.x

Cited by 5 publications

(9 citation statements)

References 36 publications

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“…We chose to use C. necator H16 because it is a nonpathogenic organism that is easy to cultivate (e.g., see reference 54) and because it showed the same phenotype as Burkholderia sp. strain N-APS2, with which the project was initiated (35).…”

Section: Resultsmentioning

confidence: 99%

“…The sodium salt of 3-sulfopropanoate was synthesized from propanoic acid and sulfonyl chloride in the presence of the radical starter azoisobutyronitrile as described previously (35). The bisulfite addition complex of 3-sulfopropanal was generated (19,29,55), but it was not a substrate for the 3-sulfopropanal dehydrogenase, and we could not convert it to the free aldehyde by published methods (23,29,55).…”

Section: Methodsmentioning

confidence: 99%

“…Homotaurine, GABA, glutamate, alanine, glycine, and 2-aminobutyrate were determined after derivatization with 2,4-dinitrofluorobenzene (DNFB) (44) and separation by reversed-phase high-pressure liquid chromatography as described previously (35). 3-Sulfopropanal and other aldehydes were separated by reversed-phase high-pressure liquid chromatography after derivatization with 2-(diphenylacetyl)indane-1,3-dione-1-hydrazone (DIH) as detailed elsewhere (35). Alternatively, a colorimetric assay after derivatization with o-aminobenzaldehyde was used to detect 3-sulfopropanal (and other aldehydes) with succinate semialdehyde as the standard (53).…”

Section: Methodsmentioning

confidence: 99%

“…Homotaurine:2-oxoglutarate aminotransferase activity was quantified by the discontinuous assay of glutamate formation (or homotaurine depletion), with reaction conditions described elsewhere (35). Kinetic constants were determined with reaction mixtures containing various homotaurine or FIG.…”

Section: Methodsmentioning

confidence: 99%

“…1A) is a natural product of marine red algae (26,36), a component (sometimes derivatized) of accredited and experimental pharmaceuticals (e.g., see reference 49), an over-the-counter "memory enhancer," and a moiety of Good buffer CAPS [3-(cyclohexylamino)-1-propanesulfonate]. The enrichment and isolation of organisms to degrade homotaurine yielded terrestrial bacteria (35); largely of relevance here is the utilization of the nitrogen moiety and excretion of sulfopropanoate. One of the isolates was identified as Cupriavidus sp.…”

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confidence: 99%

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Homotaurine Metabolized to 3-Sulfopropanoate in Cupriavidus necator H16: Enzymes and Genes in a Patchwork Pathway

Mayer

Cook

2009

J Bacteriol

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Homotaurine (3-aminopropanesulfonate), a natural product and an analogue of GABA (4-aminobutyrate), was found to be a sole source of nitrogen for Cupriavidus necator (Ralstonia eutropha) H16, whose genome sequence is known. Homotaurine nitrogen was assimilated into cell material, and the quantitative fate of the organosulfonate was sulfopropanoate, which was recovered in the growth medium. The first scalar reaction was shown to be inducible homotaurine:2-oxoglutarate aminotransferase, which released 3-sulfopropanal from homotaurine. This aminotransferase was purified to homogeneity and characterized. Peptide mass fingerprinting yielded locus tag H16_B0981, which was annotated gabT, for GABA transaminase (EC 2.6.1.19). Inducible, NAD(P) ؉ -coupled 3-sulfopropanal dehydrogenase, which yielded 3-sulfopropanoate from 3-sulfopropanal, was also purified and characterized. Peptide mass fingerprinting yielded locus tag H16_B0982, which was annotated gabD1, for succinate-semialdehyde dehydrogenase (EC 1.2.1.16). GabT and GabD1 were each induced during growth with GABA, and cotranscription of gabTD was observed. In other organisms, regulator GabC or GabR is encoded contiguous with gabTD: candidate GabR was found in strain H16 and in many other organisms. An orthologue of the GABA permease (GabP), established in Escherichia coli, is present at H16_B1890, and it was transcribed constitutively. We presume that GabRPTD are responsible for the inducible metabolism of homotaurine to intracellular 3-sulfopropanoate. The nature of the exporter of this highly charged compound was unclear until we realized from the sodium dodecyl sulfate-polyacrylamide gel electrophoresis data that sulfoacetaldehyde acetyltransferase (EC 2.3.3.15; H16_B1872) was strongly induced during growth with homotaurine and inferred that the sulfite exporter encoded at the end of the gene cluster (H16_B1874) has a broad substrate range that includes 3-sulfopropanoate.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Homotaurine Metabolized to 3-Sulfopropanoate in Cupriavidus necator H16: Enzymes and Genes in a Patchwork Pathway

Mayer

Cook

2009

J Bacteriol

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Die hormonale Steuerung der geschlechtlichen und der Fortpflanzungsvorgänge

Seitz

1939

Wachstum, Geschlecht Und Fortpflanzung

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(R)-Cysteate-nitrogen assimilation by Cupriavidus necator H16 with excretion of 3-sulfolactate: a patchwork pathway

et al. 2012

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Cupriavidus necator H16 grew exponentially with (R)-cysteate, a structural analogue of aspartate, as sole source of nitrogen in succinate-salts medium. Utilization of cysteate was quantitative and concomitant with growth and with the excretion of the deaminated product (R)-sulfolactate, which was identified thoroughly. The deaminative pathway started with transport of (R)-cysteate into the cell, which we attributed to an aspartate transporter. Transamination to sulfopyruvate involved an aspartate/(R)-cysteate:2-oxoglutarate aminotransferase (Aoa/Coa) and regeneration of the amino group acceptor by NADP⁺-coupled glutamate dehydrogenase. Reduction of sulfopyruvate to (R)-sulfolactate was catalyzed by a (S)-malate/(R)-sulfolactate dehydrogenase (Mdh/Sdh). Excretion of the sulfolactate could be attributed to the sulfite/organosulfonate exporter TauE, which was co-encoded and co-expressed, with sulfoacetaldehyde acetyltransferase (Xsc), though Xsc was irrelevant to the current pathway. The metabolic enzymes could be assayed biochemically. Aoa/Coa and Mdh/Sdh were highly enriched by protein separation, partly characterized, and the relevant locus-tags identified by peptide-mass fingerprinting. Finally, RT-PCR was used to confirm the transcription of all appropriate genes. We thus demonstrated that Cupriavidus necator H16 uses a patchwork pathway by recruitment of 'housekeeping' genes and sulfoacetaldehyde-degradative genes to scavenge for (R)-cysteate-nitrogen.

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Assimilation of homotaurine-nitrogen byBurkholderiasp. and excretion of sulfopropanoate

Cited by 5 publications

References 36 publications

Homotaurine Metabolized to 3-Sulfopropanoate in Cupriavidus necator H16: Enzymes and Genes in a Patchwork Pathway

Homotaurine Metabolized to 3-Sulfopropanoate in Cupriavidus necator H16: Enzymes and Genes in a Patchwork Pathway

Die hormonale Steuerung der geschlechtlichen und der Fortpflanzungsvorgänge

(R)-Cysteate-nitrogen assimilation by Cupriavidus necator H16 with excretion of 3-sulfolactate: a patchwork pathway

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