A Cysteine Desulfhydrase Specific for ᴅ-Cysteine from the Green Alga Chlorella fusca

Schmidt, Ahlert; Erdle, Ingrid

doi:10.1515/znc-1983-5-616

Cited by 23 publications

(9 citation statements)

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“…In several organisms, D-cysteine desulfhydrase activity (EC 4.4.1.15), decomposing D-cysteine into pyruvate, H 2 S, and ammonium, has been measured (Nagasawa et al, 1985(Nagasawa et al, , 1988. Similar activity was detected in several plant species, such as Spinacia oleracea, Chlorella fusca, Cucurbita pepo, Cucumis sativus and in suspension cultures of Nicotiana tabacum (Schmidt, 1982;Schmidt and Erdle, 1983;Rennenberg, 1983;Rennenberg et al, 1987). In all publications cited, the D-cysteine desulfhydrase activity could be clearly separated from L-cysteine desulfhydrase activity, by demonstrating different pH optima for enzyme activity (Schmidt, 1982), different sensitivity to inhibitors (Rennenberg et al, 1987), and different localizations in the cell (Rennenberg et al, 1987).…”

Section: Identification and Characterization Of Cysteine Desulfhydrasmentioning

confidence: 86%

Characterization of Cysteine‐Degrading and H₂S‐Releasing Enzymes of Higher Plants ‐ From the Field to the Test Tube and Back

Papenbrock

Riemenschneider

Kamp³

et al. 2007

Plant Biology

179

103

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Due to the clean air acts and subsequent reduction of emission of gaseous sulfur compounds sulfur deficiency became one of the major nutrient disorders in Northern Europe. Typical sulfur deficiency symptoms can be diagnosed. Especially plants of the Cruciferae family are more susceptible against pathogen attack. Sulfur fertilization can in part recover or even increase resistance against pathogens in comparison to sulfur-deficient plants. The term sulfur-induced resistance (SIR) was introduced, however, the molecular basis for SIR is largely unknown. There are several sulfur-containing compounds in plants which might be involved in SIR, such as high levels of thiols, glucosinolates, cysteine-rich proteins, phytoalexins, elemental sulfur, or H2S. Probably more than one strategy is used by plants. Species- or even variety-dependent differences in the development of SIR are probably used. Our research focussed mainly on the release of H2S as defence strategy. In field experiments using different BRASSICA NAPUS genotypes it was shown that the genetic differences among BRASSICA genotypes lead to differences in sulfur content and L-cysteine desulfhydrase activity. Another field experiment demonstrated that sulfur supply and infection with PYRENOPEZIZA BRASSICA influenced L-cysteine desulfhydrase activity in BRASSICA NAPUS. Cysteine-degrading enzymes such as cysteine desulfhydrases are hypothesized to be involved in H2S release. Several L- and D-cysteine-specific desulfhydrase candidates have been isolated and partially analyzed from the model plant ARABIDOPSIS THALIANA. However, it cannot be excluded that H2S is also released in a partial back reaction of O-acetyl-L-serine(thiol)lyase or enzymes not yet characterized. For the exact determination of the H2S concentration in the cell a H2S-specific microsensor was used the first time for plant cells. The transfer of the results obtained for application back on BRASSICA was initiated.

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Section: Identification and Characterization Of Cysteine Desulfhydrasmentioning

confidence: 86%

Characterization of Cysteine‐Degrading and H₂S‐Releasing Enzymes of Higher Plants ‐ From the Field to the Test Tube and Back

Papenbrock

Riemenschneider

Kamp³

et al. 2007

Plant Biology

179

103

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“…Green Alga and Plant D-Cysteine Desulfhydrases-D-Cysteine desulfhydrase activity was early identified in the green alga C. fusca and in the plant S. oleracea (2,3) Cyanobacteria can also utilize D-cysteine as their sole sulfur source. From this property, it was suggested that these bacteria contain a D-cysteine desulfhydrase (3).…”

Section: Comparison Of E Coli Desulfhydrase With Related Enzymesmentioning

confidence: 99%

Role of d-Cysteine Desulfhydrase in the Adaptation of Escherichia coli to d-Cysteine

Soutourina

Blanquet

Plateau

2001

Journal of Biological Chemistry

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Escherichia coli contains an enzyme activity capable of catalyzing the transformation of D-cysteine into pyruvate, H 2 S, and NH 3 (1). This enzyme was named D-cysteine desulfhydrase. A similar activity could be detected in Citrobacter freundii, Klebsiella pneumoniae, Enterobacter cloacae, Chlorella fusca, Spinacia oleracea, and various Fusobacterium species (1-4). On the other hand, no activity was found in the bacteria of genera Arthrobacter, Alcaligenes, Agrobacterium, Bacillus, Brevibacterium, Corynebacterium, Erwinia, Flavobacterium, Micrococcus, Proteus, Pseudomonas, Salmonella, Sarcina, Serratia The physiological role of bacterial D-cysteine desulfhydrase is unknown. However, recent studies indicated that E. coli growth is impaired in the presence of micromolar amounts of D-cysteine (6), suggesting that D-cysteine desulfhydrase has a detoxification function.In the present work, we report the cloning of the E. coli gene (yedO) encoding D-cysteine desulfhydrase activity. Involvement of the product of this gene in the adaptation of the cell to D-cysteine is established. Overexpression of the desulfhydrase gene protects E. coli against D-cysteine, whereas its inactivation renders the bacterium hypersensitive to the D-amino acid. Moreover, our data suggest that, similarly to L-cysteine (7), D-cysteine exerts its toxicity through an inhibition of threonine deaminase. Decomposition of D-cysteine by the desulfhydrase produces H 2 S, which the bacterium can utilize as a sulfur source. Accordingly, we establish that the presence of the yedO gene stimulates cell growth in the presence of D-cysteine as sole sulfur source. In agreement with an involvement of the desulfhydrase in sulfur metabolism, sulfate starvation induces expression of yedO. MATERIALS AND METHODSL-and D-Amino acids, ␤-chloro-D-alanine, NЈ,NЈ-dimethyl-p-phenylenediamine, 1-aminocyclopropane-1-carboxylate, and 2,4-dinitrophenylhydrazine were from Sigma. Pyridoxal phosphate and 2-oxobutyrate (sodium salt) were from Fluka. Rabbit muscle L-lactate dehydrogenase and NADH were from Roche Molecular Biochemicals. DEAE-Sephacel and Superdex 75 were from Amersham Pharmacia Biotech.Purification of D-Cysteine Desulfhydrase-Cells from E. coli strain K37 (Table I) were grown at 37°C in 8 liters of 2ϫ TY medium and harvested by centrifugation for 35 min at 3,000 ϫ g. All buffers used for the purification of D-cysteine desulfhydrase contained 10 M pyridoxal phosphate, 0.1 mM dithiothreitol, and 0.1 mM EDTA. The cell pellet was suspended in 0.1 M potassium phosphate (pH 7.0) at an optical density of 140 at 650 nm. Crude extract preparation, nucleic acid precipitation with streptomycin sulfate, and ammonium sulfate fractionation of proteins were performed as described previously (8), except that ammonium sulfate precipitations were at 35 and 55% saturation, instead of 50 and 80%, respectively. Then the protein pellet was dissolved in 50 ml of 0.1 M potassium phosphate (pH 7.0) and dialyzed overnight against 5 liters of the same buffer. The resulting solution was app...

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“…The Escherichia coli d ‐CDes (EC 4.1.99.4) is capable of catalysing the transformation of d ‐cysteine into pyruvate, H 2 S, and NH 3 [9,10]. A similar activity was detected in several plant species, such as Spinacia oleracea , Chlorella fusca , Cucurbita pepo , Cucumis sativus and in suspension cultures of Nicotiana tabacum [11–14]. In all publications cited, the d ‐CDes activity could be clearly separated from l ‐CDes activity by demonstrating different pH optima for the enzyme activity [11], different sensitivity to inhibitors [14], and different localization in the cell [14].…”

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confidence: 98%

Isolation and characterization of a D‐cysteine desulfhydrase protein from Arabidopsis thaliana

Riemenschneider¹,

Wegele²,

Schmidt³

et al. 2005

The FEBS Journal

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188

111

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It is well documented that, in general, amino acids are used in the l-form, and enzymes involved in their metabolism are stereospecific for the l-enantiomers. However, d-amino acids are widely distributed in living organisms [1]. Examples of the natural occurrence of d-amino acids include d-amino acid-containing natural peptide toxins [2], antibacterial diastereomeric peptides [3], and the presence of d-amino acids at high concentrations in human brain [4]. In plants d-amino acids were detected in gymnosperms as well as monoand dicotyledonous angiosperms of major plant families. Free d-amino acids in the low percentage range of 0.5-3% relative to their l-enantiomers are principle constituents of plants [5]. The functions of d-amino acids and their metabolism are largely unknown. Various pyridoxal-5¢-phosphate (PLP)-dependent enzymes that catalyse elimination and replacement reactions of amino acids have been purified and characterized [6]. In several organisms d-cysteine desulfhydrase (d-CDes) activity (EC 4.1.99.4) was measured; this enzyme decomposes d-cysteine into pyruvate, H 2 S, and NH 3 . A gene encoding a putative d-CDes protein was identified in Arabidopsis thaliana (L) Heynh. based on high homology to an Escherichia coli protein called YedO that has d-CDes activity. The deduced Arabidopsis protein consists of 401 amino acids and has a molecular mass of 43.9 kDa. It contains a pyridoxal-5¢-phosphate binding site. The purified recombinant mature protein had a K m for d-cysteine of 0.25 mm. Only d-cysteine but not l-cysteine was converted by d-CDes to pyruvate, H 2 S, and NH 3 . The activity was inhibited by aminooxy acetic acid and hydroxylamine, inhibitors specific for pyridoxal-5¢-phosphate dependent proteins, at low micromolar concentrations. The protein did not exhibit 1-aminocyclopropane-1-carboxylate deaminase activity (EC 3.5.99.7) as homologous bacterial proteins. Western blot analysis of isolated organelles and localization studies using fusion constructs with the green fluorescent protein indicated an intracellular localization of the nuclear encoded d-CDes protein in the mitochondria. d-CDes RNA levels increased with proceeding development of Arabidopsis but decreased in senescent plants; d-CDes protein levels remained almost unchanged in the same plants whereas specific d-CDes activity was highest in senescent plants. In plants grown in a 12-h light ⁄ 12-h dark rhythm d-CDes RNA levels were highest in the dark, whereas protein levels and enzyme activity were lower in the dark period than in the light indicating post-translational regulation. Plants grown under low sulfate concentration showed an accumulation of d-CDes RNA and increased protein levels, the d-CDes activity was almost unchanged. Putative in vivo functions of the Arabidopsis d-CDes protein are discussed.Abbreviations

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A Cysteine Desulfhydrase Specific for ᴅ-Cysteine from the Green Alga Chlorella fusca

Cited by 23 publications

References 0 publications

Characterization of Cysteine‐Degrading and H₂S‐Releasing Enzymes of Higher Plants ‐ From the Field to the Test Tube and Back

Characterization of Cysteine‐Degrading and H₂S‐Releasing Enzymes of Higher Plants ‐ From the Field to the Test Tube and Back

Role of d-Cysteine Desulfhydrase in the Adaptation of Escherichia coli to d-Cysteine

Isolation and characterization of a D‐cysteine desulfhydrase protein from Arabidopsis thaliana

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A Cysteine Desulfhydrase Specific for ᴅ-Cysteine from the Green Alga Chlorella fusca

Cited by 23 publications

References 0 publications

Characterization of Cysteine‐Degrading and H2S‐Releasing Enzymes of Higher Plants ‐ From the Field to the Test Tube and Back

Characterization of Cysteine‐Degrading and H2S‐Releasing Enzymes of Higher Plants ‐ From the Field to the Test Tube and Back

Role of d-Cysteine Desulfhydrase in the Adaptation of Escherichia coli to d-Cysteine

Isolation and characterization of a D‐cysteine desulfhydrase protein from Arabidopsis thaliana

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Characterization of Cysteine‐Degrading and H₂S‐Releasing Enzymes of Higher Plants ‐ From the Field to the Test Tube and Back

Characterization of Cysteine‐Degrading and H₂S‐Releasing Enzymes of Higher Plants ‐ From the Field to the Test Tube and Back