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Literature summary extracted from

  • Pu, Z.; Ji, F.; Wang, J.; Zhang, Y.; Sun, W.; Bao, Y.
    Rational design of meso-2,3-butanediol dehydrogenase by molecular dynamics simulation and experimental evaluations (2017), FEBS Lett., 591, 3402-3413 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.B20 gene budC, recombinant expression of SUMO-His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21-Gold from plasmid pET28a-SUMO-KpBDH Klebsiella pneumoniae
1.1.1.76 gene budC, recombinant expression of SUMO-His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21-Gold from plasmid pET28a-SUMO-KpBDH Klebsiella pneumoniae

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.B20 F212S site-directed mutagenesis, the mutant shows highly reduced activity compared to wild-type Klebsiella pneumoniae
1.1.1.B20 F212W site-directed mutagenesis, the mutant shows reduced activity compared to wild-type Klebsiella pneumoniae
1.1.1.B20 F212Y site-directed mutagenesis, the kcat of the mutant is enhanced 4-8fold compared to wild-type Klebsiella pneumoniae
1.1.1.B20 N146A site-directed mutagenesis, inactive mutant Klebsiella pneumoniae
1.1.1.B20 N146Q site-directed mutagenesis, the mutant shows unaltered activity compared to wild-type Klebsiella pneumoniae
1.1.1.76 F212S site-directed mutagenesis, the mutant shows highly reduced activity compared to wild-type Klebsiella pneumoniae
1.1.1.76 F212W site-directed mutagenesis, the mutant shows highly reduced activity compared to wild-type Klebsiella pneumoniae
1.1.1.76 F212Y site-directed mutagenesis, the kcat of the mutant is enhanced 4-8fold compared to wild-type Klebsiella pneumoniae
1.1.1.76 N146A site-directed mutagenesis, inactive mutant Klebsiella pneumoniae
1.1.1.76 N146Q site-directed mutagenesis, the mutant shows unaltered activity compared to wild-type Klebsiella pneumoniae

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.B20 1.23
-
 (2R,3S)-butane-2,3-diol pH 8.0, 25°C, recombinant mutant F212W Klebsiella pneumoniae
1.1.1.B20 1.69
-
 (2R,3S)-butane-2,3-diol pH 8.0, 25°C, recombinant wild-type enzyme Klebsiella pneumoniae
1.1.1.B20 1.76
-
 (2R,3S)-butane-2,3-diol pH 8.0, 25°C, recombinant mutant F212Y Klebsiella pneumoniae
1.1.1.B20 6.83
-
 (2R,3S)-butane-2,3-diol pH 8.0, 25°C, recombinant mutant N146Q Klebsiella pneumoniae
1.1.1.B20 15.57
-
 (2R,3S)-butane-2,3-diol pH 8.0, 25°C, recombinant mutant F212S Klebsiella pneumoniae
1.1.1.76 6.26
-
 (2S,3S)-butane-2,3-diol pH 8.0, 25°C, recombinant wild-type enzyme Klebsiella pneumoniae
1.1.1.76 7.45
-
 (2S,3S)-butane-2,3-diol pH 8.0, 25°C, recombinant mutant F212Y Klebsiella pneumoniae
1.1.1.76 10.2
-
 (2S,3S)-butane-2,3-diol pH 8.0, 25°C, recombinant mutant F212W Klebsiella pneumoniae
1.1.1.76 10.86
-
 (2S,3S)-butane-2,3-diol pH 8.0, 25°C, recombinant mutant N146Q Klebsiella pneumoniae
1.1.1.76 23.95
-
 (2S,3S)-butane-2,3-diol pH 8.0, 25°C, recombinant mutant F212S Klebsiella pneumoniae

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.B20 (2R,3S)-butane-2,3-diol + NAD+ Klebsiella pneumoniae
-
 (3R)-acetoin + NADH + H+
-
 r
1.1.1.B20 (2S,3S)-butane-2,3-diol + NAD+ Klebsiella pneumoniae
-
 (S)-acetoin + NADH + H+
-
 r
1.1.1.76 (2R,3S)-butane-2,3-diol + NAD+ Klebsiella pneumoniae
-
 (3R)-acetoin + NADH + H+
-
 r
1.1.1.76 (2S,3S)-butane-2,3-diol + NAD+ Klebsiella pneumoniae
-
 (S)-acetoin + NADH + H+
-
 r

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.B20 Klebsiella pneumoniae Q48436
-
-
1.1.1.76 Klebsiella pneumoniae Q48436
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.B20 recombinant SUMO-His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21-Gold by affinity chromatography, desalting gel filtration, and protease-mediated tag cleavage Klebsiella pneumoniae
1.1.1.76 recombinant SUMO-His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21-Gold by affinity chromatography, desalting gel filtration, and protease-mediated tag cleavage Klebsiella pneumoniae

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.B20 (2R,3S)-butane-2,3-diol + NAD+
-
 Klebsiella pneumoniae (3R)-acetoin + NADH + H+
-
 r
1.1.1.B20 (2R,3S)-butane-2,3-diol + NAD+ preferred substrate Klebsiella pneumoniae (3R)-acetoin + NADH + H+
-
 r
1.1.1.B20 (2S,3S)-butane-2,3-diol + NAD+
-
 Klebsiella pneumoniae (S)-acetoin + NADH + H+
-
 r
1.1.1.B20 additional information the meso-2,3-butanediol dehydrogenase from Klebsiella pneumoniae is active with meso-2,3-butanediol, but also with (2S,3S)-butane-2,3-diol (EC 1.1.1.76) converting them to (3R)-acetoin and (3S)-acetoin, respectively. Additionally the enzyme also has diacetyl reductase [(S)-acetoin forming] activity (EC 1.1.1.304) Klebsiella pneumoniae ?
-
-
1.1.1.76 (2R,3S)-butane-2,3-diol + NAD+
-
 Klebsiella pneumoniae (3R)-acetoin + NADH + H+
-
 r
1.1.1.76 (2R,3S)-butane-2,3-diol + NAD+ preferred substrate Klebsiella pneumoniae (3R)-acetoin + NADH + H+
-
 r
1.1.1.76 (2S,3S)-butane-2,3-diol + NAD+
-
 Klebsiella pneumoniae (S)-acetoin + NADH + H+
-
 r
1.1.1.76 additional information the meso-2,3-butanediol dehydrogenase from Klebsiella pneumoniae is active with meso-2,3-butanediol, but also with (2S,3S)-butane-2,3-diol converting them to (3R)-acetoin and (3S)-acetoin, respectively. Additionally the enzyme also has diacetyl reductase [(S)-acetoin forming] activity (EC 1.1.1.304) Klebsiella pneumoniae ?
-
-

Synonyms

EC Number Synonyms Comment Organism
1.1.1.B20 budC
-
 Klebsiella pneumoniae
1.1.1.B20 meso-2,3-BDH
-
 Klebsiella pneumoniae
1.1.1.B20 meso-2,3-butanediol dehydrogenase
-
 Klebsiella pneumoniae
1.1.1.76 budC
-
 Klebsiella pneumoniae

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.B20 25
-
 assay at Klebsiella pneumoniae
1.1.1.76 25
-
 assay at Klebsiella pneumoniae

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.B20 21.15
-
 (2R,3S)-butane-2,3-diol pH 8.0, 25°C, recombinant mutant F212S Klebsiella pneumoniae
1.1.1.B20 21.23
-
 (2R,3S)-butane-2,3-diol pH 8.0, 25°C, recombinant mutant F212W Klebsiella pneumoniae
1.1.1.B20 31.64
-
 (2R,3S)-butane-2,3-diol pH 8.0, 25°C, recombinant wild-type enzyme Klebsiella pneumoniae
1.1.1.B20 33.16
-
 (2R,3S)-butane-2,3-diol pH 8.0, 25°C, recombinant mutant N146Q Klebsiella pneumoniae
1.1.1.B20 112.03
-
 (2R,3S)-butane-2,3-diol pH 8.0, 25°C, recombinant mutant F212Y Klebsiella pneumoniae
1.1.1.76 0.56
-
 (2S,3S)-butane-2,3-diol pH 8.0, 25°C, recombinant mutant F212S Klebsiella pneumoniae
1.1.1.76 0.56
-
 (2S,3S)-butane-2,3-diol pH 8.0, 25°C, recombinant mutant F212W Klebsiella pneumoniae
1.1.1.76 0.81
-
 (2S,3S)-butane-2,3-diol pH 8.0, 25°C, recombinant mutant N146Q Klebsiella pneumoniae
1.1.1.76 3.51
-
 (2S,3S)-butane-2,3-diol pH 8.0, 25°C, recombinant wild-type enzyme Klebsiella pneumoniae
1.1.1.76 11.06
-
 (2S,3S)-butane-2,3-diol pH 8.0, 25°C, recombinant mutant F212Y Klebsiella pneumoniae

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.B20 8
-
 assay at Klebsiella pneumoniae
1.1.1.76 8
-
 assay at Klebsiella pneumoniae

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.B20 NAD+
-
 Klebsiella pneumoniae
1.1.1.B20 NADH
-
 Klebsiella pneumoniae
1.1.1.76 NAD+
-
 Klebsiella pneumoniae
1.1.1.76 NADH
-
 Klebsiella pneumoniae

General Information

EC Number General Information Comment Organism
1.1.1.B20 evolution the enzyme belongs to the short-chain dehydrogenases/reductases Klebsiella pneumoniae
1.1.1.B20 additional information identification of the the active tunnel of meso-2,3-BDH. The two short alpha-helices positioned away from the alpha4-helix possibly expose the hydrophobic ligand-binding cavity, gating the exit of product and cofactor from the activity pocket. AC binds in the active pocket including Ser139, Gln140, Ala141, Leu149, Tyr152, Gly183, Ile184, and Trp190. Residues Phe212 and Asn146 function as the key product-release sites. Three catalytic residues are Ser139, Tyr152, and Lys156. Docking study using the structure of meso-2,3-BDH (PDB ID 1GEG), molecular dynamics simulation Klebsiella pneumoniae
1.1.1.B20 physiological function the meso-2,3-butanediol dehydrogenase (meso-2,3-BDH) catalyzes NAD+-dependent conversion of meso-2,3-butanediol to acetoin (AC), a crucial external energy storage molecule in fermentive bacteria. The interconversion between (3R)-AC and meso-2,3-BD or (3S)-AC and (2S,3S)-2,3-BD is catalyzed by meso-2,3-butanediol dehydrogenase (meso-2,3-BDH) Klebsiella pneumoniae
1.1.1.76 evolution the enzyme belongs to the short-chain dehydrogenases/reductases Klebsiella pneumoniae
1.1.1.76 additional information identification of the the active tunnel of meso-2,3-BDH. The two short alpha-helices positioned away from the alpha4-helix possibly expose the hydrophobic ligand-binding cavity, gating the exit of product and cofactor from the activity pocket. AC binds in the active pocket including Ser139, Gln140, Ala141, Leu149, Tyr152, Gly183, Ile184, and Trp190. Residues Phe212 and Asn146 function as the key product-release sites. Three catalytic residues are Ser139, Tyr152, and Lys156. Docking study using the structure of meso-2,3-BDH (PDB ID 1GEG), molecular dynamics simulation Klebsiella pneumoniae
1.1.1.76 physiological function the meso-2,3-butanediol dehydrogenase (meso-2,3-BDH) catalyzes NAD+-dependent conversion of meso-2,3-butanediol to acetoin (AC), a crucial external energy storage molecule in fermentive bacteria. The interconversion between (3R)-AC and meso-2,3-BD or (3S)-AC and (2S,3S)-2,3-BD is catalyzed by meso-2,3-butanediol dehydrogenase (meso-2,3-BDH) Klebsiella pneumoniae

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.1.B20 1.36
-
 (2R,3S)-butane-2,3-diol pH 8.0, 25°C, recombinant mutant F212S Klebsiella pneumoniae
1.1.1.B20 4.86
-
 (2R,3S)-butane-2,3-diol pH 8.0, 25°C, recombinant mutant N146Q Klebsiella pneumoniae
1.1.1.B20 17.26
-
 (2R,3S)-butane-2,3-diol pH 8.0, 25°C, recombinant mutant F212W Klebsiella pneumoniae
1.1.1.B20 18.72
-
 (2R,3S)-butane-2,3-diol pH 8.0, 25°C, recombinant wild-type enzyme Klebsiella pneumoniae
1.1.1.B20 63.65
-
 (2R,3S)-butane-2,3-diol pH 8.0, 25°C, recombinant mutant F212Y Klebsiella pneumoniae
1.1.1.76 0.023
-
 (2S,3S)-butane-2,3-diol pH 8.0, 25°C, recombinant mutant F212S Klebsiella pneumoniae
1.1.1.76 0.055
-
 (2S,3S)-butane-2,3-diol pH 8.0, 25°C, recombinant mutant F212W Klebsiella pneumoniae
1.1.1.76 0.075
-
 (2S,3S)-butane-2,3-diol pH 8.0, 25°C, recombinant mutant N146Q Klebsiella pneumoniae
1.1.1.76 0.56
-
 (2S,3S)-butane-2,3-diol pH 8.0, 25°C, recombinant wild-type enzyme Klebsiella pneumoniae
1.1.1.76 1.49
-
 (2S,3S)-butane-2,3-diol pH 8.0, 25°C, recombinant mutant F212Y Klebsiella pneumoniae