Cysteine desulfhydrase activities of Salmonella typhimurium and Escherichia coli

Guarneros, Gabriel; Ortega, Manuel V.

doi:10.1016/0005-2744(70)90041-0

Cited by 49 publications

(35 citation statements)

References 8 publications

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“…Endogenous activities, representing sulfide found in the absence of added sulfite, were detected even in the cysG mutant, strain EB232. Perhaps this sulfide was the product of cysteine desulfhydrase activity (9,13). The same assay, but with thiosulfate instead of sulfite, was also found suitable for the assay of thiosulfate reductase.…”

Section: Resultsmentioning

confidence: 92%

Characterization of anaerobic sulfite reduction by Salmonella typhimurium and purification of the anaerobically induced sulfite reductase

1989

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Mutants of Salmonella typhimurium that lack the biosynthetic sulfite reductase (cysI and cysJ mutants) retain the ability to reduce sulfite for growth under anaerobic conditions (E. L. Barrett and G. W. Chang, J. Gen. Microbiol., 115:513-516, 1979). Here we report studies of sulfite reduction by a cysI mutant of S. typhimurium and purification of the associated anaerobic sulfite reductase. Sulfite reduction for anaerobic growth did not require a reducing atmosphere but was prevented by an argon atmosphere contaminated with air (<0.33%). It was also prevented by the presence of 0.1 mM nitrate, which argues against a strictly biosynthetic role for anaerobic sulfite reduction. Anaerobic growth in liquid minimal medium, but not on agar, was found to require additions of trace amounts (lo-7 M) of cysteine. Spontaneous mutants that grew under the argon contaminated with air also lost the requirement for 10-7 M cysteine for anaerobic growth in liquid. A role for sulfite reduction in anaerobic energy generation was contraindicated by the findings that sulfite reduction did not improve cell yields, and anaerobic sulfite reductase activity was greatest during the stationary phase of growth. Sulfite reductase was purified from the cytoplasmic fraction of the anaerobically grown cysI mutant and was purified 190-fold. The most effective donor in crude extracts was NADH. NADPH and methyl viologen were, respectively, 40 and 30% as effective as NADH. Oxygen reversibly inhibited the enzyme. Two highmolecular-weight proteins separated by gel filtration (Mr 360,000 and 490,000, respectively) were required for maximal activity with NADH. Indirect evidence, including in vitro complementation experiments with a cysG mutant extract, suggested that the 3609000-Mr component contains siroheme and is the terminal reductase.This component was further purified to near homogeneity and was found to consist of a single subunit of molecular weight 67,500. The anaerobic sulfite reductase showed some resemblance to the biosynthetic sulfite reductase, but apparently it has a unique, as yet unidentified function.Although much is known about sulfite reduction by the sulfate-reducing bacteria (1, 23, 28), very little is known about the anaerobic production of H2S from sulfite by members of the family Enterobacteriaceae. The ability to perform this reaction is routinely used in diagnostic laboratories to differentiate Salmonella species (H2S positive) from Escherichia coli (H2S negative). We have hypothesized that the enzymes involved may explain the observation that cysI and cysJ (biosynthetic sulfite reductase) mutants of S. typhimurium behave as prototrophs under anaerobic conditions (2), whereas E. coli cysI and cysJ mutants are auxotrophs under both aerobic and anaerobic conditions (E. L. Barrett, unpublished findings). Like E. coli, S. typhimurium mutants defective in cysG, which is required for synthesis of the siroheme prosthetic group (25), do not produce H2S from sulfite and do not behave as prototrophs under anaerobic growth conditions. The...

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Section: Resultsmentioning

confidence: 92%

Characterization of anaerobic sulfite reduction by Salmonella typhimurium and purification of the anaerobically induced sulfite reductase

1989

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“…It has been reported that hydrogen sulfide is produced from -cysteine by the enzymic action of -cysteine desulfhydrase, which catalyses the α,β-elimination of -cysteine to hydrogen sulfide, pyruvate and ammonia (Guarneros & Ortega, 1970). Since hydrogen sulfide is highly toxic for mammalian cells (Beauchamp et al, 1984) and induces the modification and release of haemoglobin in erythrocytes (Kurzban et al, 1999), enzymes participating in the production of hydrogen sulfide in the subgingival sulcus might contribute to the initiation and progression of periodontal diseases, and to abscess formation (Carlsson et al, 1993 ;Persson et al, 1990).…”

Section: Introductionmentioning

confidence: 99%

lcd from Streptococcus anginosus encodes a C-S lyase with α,β-elimination activity that degrades l-cysteine The DDBJ accession number for the Streptococcus anginosus lcd gene sequence reported in this paper is AB084812.

et al. 2002

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Hydrogen sulfide is highly toxic to mammalian cells. It has also been postulated that hydrogen sulfide modifies haemoglobin resulting in haemolysis. The enzyme that produces hydrogen sulfide from L-cysteine was purified from Streptococcus anginosus. Using the N-terminal amino acid sequence of the purified enzyme, the lcd gene encoding L-cysteine desulfhydrase was cloned ; the recombinant protein was then purified to examine its enzymic and biological characteristics. This L-cysteine desulfhydrase had the Michaelis-Menten kinetics K m l062 mM and

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“…S. enterica and E. coli have several metabolic enzymes with cysteine desulfhydrase activity, including cystathionine ␤-lyase (MetC), cysteine synthase A (CysK), cysteine synthase B (CysM), ␤-cystathionase (MalY), and tryptophanase (TnaA; E. coli only) (25,27). In contrast to E. coli, S. enterica has a dedicated cysteine desulfhydrase activity that is strongly induced in response to cysteine (26,28,29). It was recently demonstrated that this dedicated cysteine desulfhydrase (CDS; EC 2.5.1.47) is encoded by stm0458, which was renamed cdsH (25).…”

mentioning

confidence: 99%

Endogenous Synthesis of 2-Aminoacrylate Contributes to Cysteine Sensitivity in Salmonella enterica

et al. 2014

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RidA, the archetype member of the widely conserved RidA/YER057c/UK114 family of proteins, prevents reactive enamine/imine intermediates from accumulating in Salmonella enterica by catalyzing their hydrolysis to stable keto acid products. In the absence of RidA, endogenous 2-aminoacrylate persists in the cellular environment long enough to damage a growing list of essential metabolic enzymes. Prior studies have focused on the dehydration of serine by the pyridoxal 5=-phosphate (PLP)-dependent serine/threonine dehydratases, IlvA and TdcB, as sources of endogenous 2-aminoacrylate. The current study describes an additional source of endogenous 2-aminoacrylate derived from cysteine. The results of in vivo analysis show that the cysteine sensitivity of a ridA strain is contingent upon CdsH, the predominant cysteine desulfhydrase in S. enterica. The impact of cysteine on 2-aminoacrylate accumulation is shown to be unaffected by the presence of serine/threonine dehydratases, revealing another mechanism of endogenous 2-aminoacrylate production. Experiments in vitro suggest that 2-aminoacrylate is released from CdsH following cysteine desulfhydration, resulting in an unbound aminoacrylate substrate for RidA. This work expands our understanding of the role played by RidA in preventing enamine stress resulting from multiple normal metabolic processes.

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Cysteine desulfhydrase activities of Salmonella typhimurium and Escherichia coli

Cited by 49 publications

References 8 publications

Characterization of anaerobic sulfite reduction by Salmonella typhimurium and purification of the anaerobically induced sulfite reductase

Characterization of anaerobic sulfite reduction by Salmonella typhimurium and purification of the anaerobically induced sulfite reductase

lcd from Streptococcus anginosus encodes a C-S lyase with α,β-elimination activity that degrades l-cysteine The DDBJ accession number for the Streptococcus anginosus lcd gene sequence reported in this paper is AB084812.

Endogenous Synthesis of 2-Aminoacrylate Contributes to Cysteine Sensitivity in Salmonella enterica

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