Sulfate: Turbidimetric and nephelometric methods

Bacterial degradation of sulfoacetate, a widespread natural product, proceeds via sulfoacetaldehyde and requires a considerable initial energy input. Whereas the fate of sulfoacetaldehyde in Cupriavidus necator (Ralstonia eutropha) H16 is known, the pathway from sulfoacetate to sulfoacetaldehyde is not. The genome sequence of the organism enabled us to hypothesize that the inducible pathway, which initiates sau (sulfoacetate utilization), involved a four-gene cluster (sauRSTU; H16_A2746 to H16_A2749). The sauR gene, divergently orientated to the other three genes, probably encodes the transcriptional regulator of the presumed sauSTU operon, which is subject to inducible transcription. SauU was tentatively identified as a transporter of the major facilitator superfamily, and SauT was deduced to be a sulfoacetate-CoA ligase. SauT was a labile protein, but it could be separated and shown to generate AMP and an unknown, labile CoA-derivative from sulfoacetate, CoA, and ATP. This unknown compound, analyzed by MALDI-TOF-MS, had a relative molecular mass of 889.7, which identified it as protonated sulfoacetyl-CoA (calculated 889.6). SauS was deduced to be sulfoacetaldehyde dehydrogenase (acylating). The enzyme was purified 175-fold to homogeneity and characterized. Peptide mass fingerprinting confirmed the sauS locus (H16_A2747). SauS converted sulfoacetyl-CoA and NADPH to sulfoacetaldehyde, CoA, and NADP ؉ , thus confirming the hypothesis.

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“…Sulfate was quantified turbidimetrically as an insoluble suspension of BaSO 4 (29). Sulfite was quantified as the fuchsin adduct (30).…”

Section: Methodsmentioning

confidence: 99%

Sulfoacetate Is Degraded via a Novel Pathway Involving Sulfoacetyl-CoA and Sulfoacetaldehyde in Cupriavidus necator H16

Weinitschke

Hollemeyer

Kusian

et al. 2010

Journal of Biological Chemistry

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“…Reversed-phase chromatography was used to quantify taurine (Laue et al 1997) or sulfoacetaldehyde (Cunningham et al 1998) after derivatization. Sulfate was determined turbidimetrically as a suspension of BaSO 4 (Sörbo 1987). Ammonium ion was assayed colorimetrically by the Berthelot reaction (Gesellschaft Deutscher Chemiker 1996).…”

Section: Methodsmentioning

confidence: 99%

Sulfoacetate generated by Rhodopseudomonas palustris from taurine

et al. 2004

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Genes thought to encode (a) the regulator of taurine catabolism under carbon-limiting or nitrogen-limiting conditions and (b) taurine dehydrogenase were found in the genome of Rhodopseudomonas palustris. The organism utilized taurine quantitatively as a sole source of nitrogen (but not of carbon) for aerobic and photoheterotrophic growth. No sulfate was released, and the C-sulfonate bond was recovered stoichiometrically as sulfoacetate, which was identified by mass spectrometry. An inducible sulfoacetaldehyde dehydrogenase was detected. R. palustris thus contains a pathway to generate a natural product that was previously believed to be formed solely from sulfoquinovose.

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“…Sulfate was occasionally assayed as the suspended barium salt (Sörbo 1987). Sulfite was routinely determined as the fuchsin complex (Kondo et al 1982) and occasionally examined by ion chromatography (Laue et al 1996).…”

Section: Methodsmentioning

confidence: 99%

Ethanedisulfonate is degraded via sulfoacetaldehyde in Ralstonia sp. strain EDS1

Denger

Cook

2001

Archives of Microbiology

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Aerobic enrichment cultures (11) yielded three cultures able to utilise ethane-1,2-disulfonate as sole source of carbon and energy in salts medium. Two pure cultures were obtained and we worked with strain EDS1, which was assigned to the genus Ralstonia on the basis of its 16S rDNA sequence and simple taxonomic tests. Strain EDS1 utilised at least seven alkane(di)sulfonates, ethane-1,2-disulfonate, taurine, isethionate, sulfoacetate, sulfoacetaldehyde and propane-1,3-disulfonate, as well as methanesulfonate and formate. Growth with ethanedisulfonate was concomitant with substrate disappearance and the formation of 2 mol sulfate per mol substrate. The growth yield, 7 g protein (mol C) -1 , indicated quantitative utilisation of the substrate. Ethanedisulfonate-dependent oxygen uptake of whole cells during growth rose to a maximum before the end of growth and then sank rapidly; this was interpreted as evidence for an inducible desulfonative oxygenase that was not active in cell extracts. Inducible sulfoacetaldehyde sulfo-lyase was detected at high activity. Inducible degradation of taurine or isethionate or sulfoacetate via sulfoacetaldehyde sulfo-lyase is interpreted from the data.

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Sulfate: Turbidimetric and nephelometric methods

Cited by 134 publications

References 15 publications

Sulfoacetate Is Degraded via a Novel Pathway Involving Sulfoacetyl-CoA and Sulfoacetaldehyde in Cupriavidus necator H16

Sulfoacetate Is Degraded via a Novel Pathway Involving Sulfoacetyl-CoA and Sulfoacetaldehyde in Cupriavidus necator H16

Sulfoacetate generated by Rhodopseudomonas palustris from taurine

Ethanedisulfonate is degraded via sulfoacetaldehyde in Ralstonia sp. strain EDS1

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