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Literature summary extracted from

  • Farsi, A.; Lodha, P.H.; Skanes, J.E.; Los, H.; Kalidindi, N.; Aitken, S.M.
    Interconversion of a pair of active-site residues in Escherichia coli cystathionine gamma-synthase, E. coli cystathionine beta-lyase, and Saccharomyces cerevisiae cystathionine gamma-lyase and development of tools for the investigation of their mechanisms (2009), Biochem. Cell Biol., 87, 445-457.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.4.1.1 the His-tagged enzyme is expressed in Escherichia coli KS1000 cells Saccharomyces cerevisiae
4.4.1.13 mutant enzymes are expressed in Escherichia coli KS1000 cells Escherichia coli

Protein Variants

EC Number Protein Variants Comment Organism
2.5.1.48 D45F reduction in catalytic effciency, without change in reaction specificity Escherichia coli
2.5.1.48 D45F/E325Y reduction in catalytic effciency, without change in reaction specificity Escherichia coli
2.5.1.48 E325Y reduction in catalytic effciency, without change in reaction specificity Escherichia coli
4.4.1.1 E333A the mutation leads to 30fold reduction of kcat/Km for L-cystathionine Saccharomyces cerevisiae
4.4.1.1 E333Y the mutation leads to 30fold reduction of kcat/Km for L-cystathionine Saccharomyces cerevisiae
4.4.1.1 E48A the mutation leads to about 6fold reduction of kcat/Km for L-cystathionine Saccharomyces cerevisiae
4.4.1.1 E48D the mutation leads to about 5fold reduction of kcat/Km for L-cystathionine Saccharomyces cerevisiae
4.4.1.1 E48F the mutation leads to about 9fold reduction of kcat/Km for L-cystathionine Saccharomyces cerevisiae
4.4.1.13 F55D the mutant shows 74fold reduced catalytic efficiency compared to the wild type enzyme Escherichia coli
4.4.1.13 F55D/Y338E the mutant shows 58000fold reduced catalytic efficiency compared to the wild type enzyme Escherichia coli
4.4.1.13 Y338E the mutant shows 2850fold reduced catalytic efficiency compared to the wild type enzyme Escherichia coli

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.5.1.48 0.11
-
 L-cystathionine wild-type, 25°C, pH 7.8 Escherichia coli
2.5.1.48 0.26
-
 L-cystathionine mutant E325Y 25°C, pH 7.8 Escherichia coli
2.5.1.48 1.3
-
 O-succinyl-L-homoserine wild-type, gamma-elimination, 25°C, pH 7.8 Escherichia coli
2.5.1.48 1.5
-
 O-succinyl-L-homoserine mutant D45F/E325Y, gamma-elimination, 25°C, pH 7.8 Escherichia coli
2.5.1.48 1.7
-
 O-succinyl-L-homoserine mutant E325Y, gamma-elimination, 25°C, pH 7.8 Escherichia coli
2.5.1.48 2
-
 O-succinyl-L-homoserine mutant D45F, gamma-elimination, 25°C, pH 7.8 Escherichia coli
2.5.1.48 2.1
-
 O-succinyl-L-homoserine mutant E325Y, gamma-replacement, 25°C, pH 7.8 Escherichia coli
2.5.1.48 2.4
-
 L-cystathionine mutant D45F, 25°C, pH 7.8 Escherichia coli
2.5.1.48 2.5
-
 O-succinyl-L-homoserine wild-type, gamma-replacement, 25°C, pH 7.8 Escherichia coli
2.5.1.48 5.4
-
 O-succinyl-L-homoserine mutant D45F, gamma-replacement, 25°C, pH 7.8 Escherichia coli
4.4.1.1 0.42
-
 L-cystathionine His-tagged recombinant wild type enzyme, in 50 mM potassium phosphate, pH 7.8, at 25°C Saccharomyces cerevisiae
4.4.1.1 0.45
-
 L-cystathionine native wild type enzyme, in 50 mM potassium phosphate, pH 7.8, at 25°C Saccharomyces cerevisiae
4.4.1.1 0.81
-
 L-cystathionine mutant enzyme E48F, in 50 mM potassium phosphate, pH 7.8, at 25°C Saccharomyces cerevisiae
4.4.1.1 0.82
-
 L-cystathionine mutant enzyme E48A, in 50 mM potassium phosphate, pH 7.8, at 25°C Saccharomyces cerevisiae
4.4.1.1 1.11
-
 L-cystathionine mutant enzyme E48D, in 50 mM potassium phosphate, pH 7.8, at 25°C Saccharomyces cerevisiae
4.4.1.1 5.2
-
 L-cystathionine mutant enzyme E333A, in 50 mM potassium phosphate, pH 7.8, at 25°C Saccharomyces cerevisiae

Organism

EC Number Organism UniProt Comment Textmining
2.5.1.48 Escherichia coli
-
-
-
4.4.1.1 Saccharomyces cerevisiae
-
-
-
4.4.1.13 Escherichia coli
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.4.1.1 ammonium sulfate precipitation, butyl-Toyopearl 650M resin column chromatography, and DEAE column chromatography Saccharomyces cerevisiae
4.4.1.13 ammonium sulfate precipitation, Toyopearl butyl-650M resin column chromatography, and DEAE column chromatography (untagged enzyme), or Ni-NTA column chromatography (His-tagged enzyme) Escherichia coli

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.5.1.48 L-cystathionine + acetate
-
 Escherichia coli O-acetyl-L-homoserine + L-cysteine
-
 r
2.5.1.48 O-acetyl-L-homoserine + L-cysteine gamma-replacement Escherichia coli L-cystathionine + acetate
-
 r
2.5.1.48 O-succinyl-L-homoserine + H2O gamma-elimination Escherichia coli succinate + 2-oxobutyrate + NH3
-
 ?
4.4.1.1 L-cystathionine + H2O
-
 Saccharomyces cerevisiae L-homocysteine + pyruvate + NH3
-
 ?
4.4.1.13 L-cystathionine + H2O
-
 Escherichia coli L-homocysteine + NH3 + pyruvate
-
 ?

Synonyms

EC Number Synonyms Comment Organism
4.4.1.1 cgl
-
 Saccharomyces cerevisiae
4.4.1.13 CBL
-
 Escherichia coli

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.5.1.48 1.23
-
 O-succinyl-L-homoserine mutant D45F/E325Y, gamma-elimination, 25°C, pH 7.8 Escherichia coli
2.5.1.48 1.39
-
 O-succinyl-L-homoserine mutant E325Y, gamma-elimination, 25°C, pH 7.8 Escherichia coli
2.5.1.48 1.4
-
 O-succinyl-L-homoserine mutant D45F, gamma-elimination, 25°C, pH 7.8 Escherichia coli
2.5.1.48 1.8
-
 O-succinyl-L-homoserine wild-type, gamma-elimination, 25°C, pH 7.8 Escherichia coli
2.5.1.48 5.6
-
 L-cystathionine mutant D45F, 25°C, pH 7.8 Escherichia coli
2.5.1.48 11.1
-
 L-cystathionine mutant E325Y 25°C, pH 7.8 Escherichia coli
2.5.1.48 121
-
 L-cystathionine wild-type, 25°C, pH 7.8 Escherichia coli
4.4.1.1 0.39
-
 L-cystathionine mutant enzyme E48F, in 50 mM potassium phosphate, pH 7.8, at 25°C Saccharomyces cerevisiae
4.4.1.1 0.67
-
 L-cystathionine mutant enzyme E48A, in 50 mM potassium phosphate, pH 7.8, at 25°C Saccharomyces cerevisiae
4.4.1.1 0.69
-
 L-cystathionine mutant enzyme E333A, in 50 mM potassium phosphate, pH 7.8, at 25°C Saccharomyces cerevisiae
4.4.1.1 0.76
-
 L-cystathionine mutant enzyme E48D, in 50 mM potassium phosphate, pH 7.8, at 25°C Saccharomyces cerevisiae
4.4.1.1 0.9
-
 L-cystathionine native wild type enzyme, in 50 mM potassium phosphate, pH 7.8, at 25°C Saccharomyces cerevisiae
4.4.1.1 1.81
-
 L-cystathionine His-tagged recombinant wild type enzyme, in 50 mM potassium phosphate, pH 7.8, at 25°C Saccharomyces cerevisiae

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
4.4.1.1 7 8
-
 Saccharomyces cerevisiae

Cofactor

EC Number Cofactor Comment Organism Structure
4.4.1.1 pyridoxal 5'-phosphate
-
 Saccharomyces cerevisiae
4.4.1.13 pyridoxal 5'-phosphate
-
 Escherichia coli

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.5.1.48 0.69
-
 O-succinyl-L-homoserine mutant D45F, gamma-elimination, 25°C, pH 7.8 Escherichia coli
2.5.1.48 0.8
-
 O-succinyl-L-homoserine mutant D45F/E325Y, gamma-elimination, 25°C, pH 7.8 Escherichia coli
2.5.1.48 0.8
-
 O-succinyl-L-homoserine mutant E325Y, gamma-elimination, 25°C, pH 7.8 Escherichia coli
2.5.1.48 1.35
-
 O-succinyl-L-homoserine wild-type, gamma-elimination, 25°C, pH 7.8 Escherichia coli
2.5.1.48 2
-
 L-cystathionine mutant D45F, 25°C, pH 7.8 Escherichia coli
2.5.1.48 3
-
 O-succinyl-L-homoserine mutant D45F, gamma-replacement, 25°C, pH 7.8 Escherichia coli
2.5.1.48 7
-
 O-succinyl-L-homoserine mutant E325Y, gamma-replacement, 25°C, pH 7.8 Escherichia coli
2.5.1.48 41
-
 L-cystathionine mutant E325Y 25°C, pH 7.8 Escherichia coli
2.5.1.48 49
-
 O-succinyl-L-homoserine wild-type, gamma-replacement, 25°C, pH 7.8 Escherichia coli
2.5.1.48 1060
-
 L-cystathionine wild-type, 25°C, pH 7.8 Escherichia coli
4.4.1.1 0.132
-
 L-cystathionine mutant enzyme E333A, in 50 mM potassium phosphate, pH 7.8, at 25°C Saccharomyces cerevisiae
4.4.1.1 0.147
-
 L-cystathionine mutant enzyme E333Y, in 50 mM potassium phosphate, pH 7.8, at 25°C Saccharomyces cerevisiae
4.4.1.1 0.49
-
 L-cystathionine mutant enzyme E48F, in 50 mM potassium phosphate, pH 7.8, at 25°C Saccharomyces cerevisiae
4.4.1.1 0.68
-
 L-cystathionine mutant enzyme E48D, in 50 mM potassium phosphate, pH 7.8, at 25°C Saccharomyces cerevisiae
4.4.1.1 0.82
-
 L-cystathionine mutant enzyme E48A, in 50 mM potassium phosphate, pH 7.8, at 25°C Saccharomyces cerevisiae
4.4.1.1 1.98
-
 L-cystathionine native wild type enzyme, in 50 mM potassium phosphate, pH 7.8, at 25°C Saccharomyces cerevisiae
4.4.1.1 4.3
-
 L-cystathionine His-tagged recombinant wild type enzyme, in 50 mM potassium phosphate, pH 7.8, at 25°C Saccharomyces cerevisiae