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Literature summary extracted from

  • Carsten, J.M.; Philipp, A.; Sieber, V.
    Characterization of recombinantly expressed dihydroxy-acid dehydratase from Sulfobus solfataricus - a key enzyme for the conversion of carbohydrates into chemicals (2015), J. Biotechnol., 211, 31-41.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
4.2.1.9 2-mercaptoethanol increases activity up to 3fold. Activation modulates thermotolerance at 50°C Saccharolobus solfataricus
4.2.1.9 additional information heat activation: the heat treatment step in purification at 80°C increases the total activity about 2.5 to 3-fold and the specific activity about 20 to 30-fold Saccharolobus solfataricus

Application

EC Number Application Comment Organism
4.2.1.9 synthesis the enzyme can be used for a convenient one-step synthesis route from D-gluconate to 2-dehydro-3-deoxy-D-gluconate Saccharolobus solfataricus

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.2.1.9 expression in Escherichia coli BL21 (DE3) Saccharolobus solfataricus

Inhibitors

EC Number Inhibitors Comment Organism Structure
4.2.1.9 (NH4)2Fe(SO4)2 10 mM, 50°C, 45% loss of activity, dehydration of D-gluconate Saccharolobus solfataricus
4.2.1.9 BaCl2 100 mM, 50°C, 43% loss of activity, dehydration of D-gluconate Saccharolobus solfataricus
4.2.1.9 butanol
-
 Saccharolobus solfataricus
4.2.1.9 CaCl2 10 mM, 50°C, 77% loss of activity, dehydration of D-gluconate Saccharolobus solfataricus
4.2.1.9 CuSO4 10 mM, 50°C, 86% loss of activity, dehydration of D-gluconate Saccharolobus solfataricus
4.2.1.9 D-glycerate competitive. Presence of 20 mM D-glycerate decreases catalytic activity towards D-gluconate drastically to 5% residual activity Saccharolobus solfataricus
4.2.1.9 ethanol
-
 Saccharolobus solfataricus
4.2.1.9 imidazole leads to a complete elimination of enzyme activity towards D-gluconate Saccharolobus solfataricus
4.2.1.9 isobutanol
-
 Saccharolobus solfataricus
4.2.1.9 ZnCl2 10 mM, 50°C, 95% loss of activity, dehydration of D-gluconate Saccharolobus solfataricus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.2.1.9 2.1
-
 2,3-Dihydroxy-3-methylbutanoate pH 7, 50°C Saccharolobus solfataricus
4.2.1.9 7.8
-
 D-gluconate pH 7, 50°C Saccharolobus solfataricus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
4.2.1.9 Fe-S center
-
 Saccharolobus solfataricus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4.2.1.9 2,3-dihydroxy-3-methylbutanoate Saccharolobus solfataricus biosynthesis pathway of the branched chain amino acids valine and isoleucine 3-methyl-2-oxobutanoate + H2O
-
 ?
4.2.1.9 2,3-dihydroxy-3-methylbutanoate Saccharolobus solfataricus P2 biosynthesis pathway of the branched chain amino acids valine and isoleucine 3-methyl-2-oxobutanoate + H2O
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
4.2.1.9 Saccharolobus solfataricus Q97UB2
-
-
4.2.1.9 Saccharolobus solfataricus P2 Q97UB2
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.2.1.9
-
 Saccharolobus solfataricus

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
4.2.1.9 10.7
-
 pH 7, 50°C, conversion of D-glycerate to pyruvate Saccharolobus solfataricus

Storage Stability

EC Number Storage Stability Organism
4.2.1.9 -80°C, after purification of His-tagged enzyme the total enzyme activity is decreased to 43% of the crude extract and even further to 17% when stored at -80 °C for one day Saccharolobus solfataricus
4.2.1.9 -80°C, specific activity for D-gluconate of the frozen/thawed enzyme is stable for 60 min, but loses about 70% after a maximum of 100 min Saccharolobus solfataricus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.2.1.9 2,3-dihydroxy-3-methylbutanoate
-
 Saccharolobus solfataricus 3-methyl-2-oxobutanoate + H2O
-
 ?
4.2.1.9 2,3-dihydroxy-3-methylbutanoate biosynthesis pathway of the branched chain amino acids valine and isoleucine Saccharolobus solfataricus 3-methyl-2-oxobutanoate + H2O
-
 ?
4.2.1.9 2,3-dihydroxy-3-methylbutanoate
-
 Saccharolobus solfataricus P2 3-methyl-2-oxobutanoate + H2O
-
 ?
4.2.1.9 2,3-dihydroxy-3-methylbutanoate biosynthesis pathway of the branched chain amino acids valine and isoleucine Saccharolobus solfataricus P2 3-methyl-2-oxobutanoate + H2O
-
 ?
4.2.1.9 D-gluconate
-
 Saccharolobus solfataricus 2-dehydro-3-deoxy-D-gluconate + H2O
-
 ?
4.2.1.9 D-gluconate
-
 Saccharolobus solfataricus P2 2-dehydro-3-deoxy-D-gluconate + H2O
-
 ?
4.2.1.9 D-glycerate the enzyme catalyzes the non-natural dehydration of D-glycerate to pyruvate Saccharolobus solfataricus pyruvate + H2O
-
 ?
4.2.1.9 D-glycerate the enzyme catalyzes the non-natural dehydration of D-glycerate to pyruvate Saccharolobus solfataricus P2 pyruvate + H2O
-
 ?

Synonyms

EC Number Synonyms Comment Organism
4.2.1.9 SsDHAD
-
 Saccharolobus solfataricus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.2.1.9 77
-
 dehydration of D-gluconate Saccharolobus solfataricus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
4.2.1.9 50
-
 half-life: 1028 min Saccharolobus solfataricus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.2.1.9 0.011
-
 D-glycerate pH 7, 50°C, non-activated enzyme Saccharolobus solfataricus
4.2.1.9 0.03
-
 D-glycerate pH 7, 50°C, enzyme activated by 2-mercaptoethanol Saccharolobus solfataricus
4.2.1.9 0.31
-
 2,3-Dihydroxy-3-methylbutanoate pH 7, 50°C, enzyme activated by 2-mercaptoethanol Saccharolobus solfataricus
4.2.1.9 0.31
-
 2,3-Dihydroxy-3-methylbutanoate pH 7, 50°C, non-activated enzyme Saccharolobus solfataricus
4.2.1.9 0.4
-
 D-gluconate pH 7, 50°C, non-activated enzyme Saccharolobus solfataricus
4.2.1.9 1.19
-
 D-gluconate pH 7, 50°C, enzyme activated by 2-mercaptoethanol Saccharolobus solfataricus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
4.2.1.9 6.2
-
 dehydration of D-gluconate Saccharolobus solfataricus

pH Range

EC Number pH Minimum pH Maximum Comment Organism
4.2.1.9 5.2 8.2 pH 5.2: about 40% of maximal activity, pH 8.2: about 40% of maximal activity, dehydration of D-gluconate Saccharolobus solfataricus

Cofactor

EC Number Cofactor Comment Organism Structure
4.2.1.9 Fe-S center
-
 Saccharolobus solfataricus

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
4.2.1.9 2.3
-
 D-glycerate 50°C, pH 7 Saccharolobus solfataricus

IC50 Value

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
4.2.1.9 additional information
-
 IC50: 15% (v/v) Saccharolobus solfataricus ethanol
4.2.1.9 additional information
-
 IC50: 4% (v/v) Saccharolobus solfataricus isobutanol
4.2.1.9 additional information
-
 IC50: 4.5% (v/v) Saccharolobus solfataricus butanol

General Information

EC Number General Information Comment Organism
4.2.1.9 metabolism biosynthesis pathway of the branched chain amino acids valine and isoleucine Saccharolobus solfataricus