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Literature summary extracted from

  • Maddock, D.J.; Patrick, W.M.; Gerth, M.L.
    Substitutions at the cofactor phosphate-binding site of a clostridial alcohol dehydrogenase lead to unexpected changes in substrate specificity (2015), Protein Eng. Des. Sel., 28, 251-258.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.4 expression in Escherichia coli Clostridium autoethanogenum

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.4 G198D/S199 V/P201E/Y218A mutant displays no detectable activity with NADPH as cofactor, but uses NADH with kcat/Km value of 12 per s and mM Clostridium autoethanogenum
1.1.1.4 additional information variation of residue S199 does not yield any NADH-dependent enzymes Clostridium autoethanogenum
1.1.1.4 S199A increase in catalytic efficiencies Clostridium autoethanogenum
1.1.1.4 S199C decrease in catalytic efficiencies, with no significant changes in substrate preference Clostridium autoethanogenum
1.1.1.4 S199G decrease in catalytic efficiencies, with no significant changes in substrate preference Clostridium autoethanogenum
1.1.1.4 S199R increase in catalytic efficiencies Clostridium autoethanogenum
1.1.1.4 S199W decrease in catalytic efficiencies, with no significant changes in substrate preference Clostridium autoethanogenum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.4 0.0017
-
 NADPH wild-type, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 0.0056
-
 NADPH mutant S199A, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 0.036
-
 NADPH mutant S199R, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 0.8
-
 2-butanone mutant S199A, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 0.9
-
 acetone mutant S199A, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 1
-
 acetone wild-type, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 1.1
-
 acetone mutant S199R, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 1.2
-
 acetone mutant S199C, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 1.5
-
 2-butanone mutant S199C, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 1.6
-
 acetone mutant S199G, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 1.8
-
 2-butanone wild-type, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 1.9
-
 2-butanone mutant S199R, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 2
-
 acetone mutant S199W, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 2.4
-
 2-butanone mutant S199G, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 3.6
-
 2-butanone mutant S199W, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 34
-
 (R)-acetoin mutant S199R, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 45
-
 (R)-acetoin mutant S199C, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 98
-
 (R)-acetoin wild-type, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 120
-
 (R)-acetoin mutant S199A, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 190
-
 (R)-acetoin mutant S199W, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 230
-
 (R)-acetoin mutant S199G, pH 7.5, 25°C Clostridium autoethanogenum

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.4 Clostridium autoethanogenum F8TEL7
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.4 (R)-acetoin + NADPH + H+
-
 Clostridium autoethanogenum (2R,3R)-2,3-butanediol + NAD+
-
 ?
1.1.1.4 2-butanone + NADP+
-
 Clostridium autoethanogenum 2-butanol + NADPH + H+
-
 ?
1.1.1.4 acetone + NADPH + H+
-
 Clostridium autoethanogenum propan-2-ol + NADP+
-
 ?

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.4 2 8 2-butanone mutant S199W, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 3 6 acetone mutant S199C, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 23
-
 2-butanone mutant S199C, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 32
-
 NADPH mutant S199R, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 37
-
 2-butanone wild-type, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 40
-
 2-butanone mutant S199G, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 41
-
 (R)-acetoin mutant S199C, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 46
-
 2-butanone mutant S199A, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 48
-
 acetone mutant S199W, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 50
-
 NADPH mutant S199A, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 50
-
 2-butanone mutant S199R, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 55
-
 acetone wild-type, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 60
-
 acetone mutant S199A, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 62
-
 acetone mutant S199G, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 66
-
 (R)-acetoin mutant S199W, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 68
-
 NADPH wild-type, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 89
-
 acetone mutant S199R, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 95
-
 (R)-acetoin mutant S199R, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 110
-
 (R)-acetoin mutant S199G, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 150
-
 (R)-acetoin wild-type, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 230
-
 (R)-acetoin mutant S199A, pH 7.5, 25°C Clostridium autoethanogenum

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.4 NADH cofoactor of mutant G198D/S199 V/P201E/Y218A Clostridium autoethanogenum
1.1.1.4 NADPH strictly dependent on NADPH Clostridium autoethanogenum

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.1.4 0.35
-
 (R)-acetoin mutant S199W, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 0.48
-
 (R)-acetoin mutant S199G, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 0.9
-
 (R)-acetoin mutant S199C, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 1.6
-
 (R)-acetoin wild-type, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 1.9
-
 (R)-acetoin mutant S199A, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 2.8
-
 (R)-acetoin mutant S199R, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 7.8
-
 2-butanone mutant S199W, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 15
-
 2-butanone mutant S199C, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 17
-
 2-butanone mutant S199G, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 21
-
 2-butanone wild-type, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 24
-
 acetone mutant S199W, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 26
-
 2-butanone mutant S199R, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 30
-
 acetone mutant S199C, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 39
-
 acetone mutant S199G, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 55
-
 acetone wild-type, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 61
-
 2-butanone mutant S199A, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 67
-
 acetone mutant S199A, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 81
-
 acetone mutant S199R, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 890
-
 NADPH mutant S199R, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 8900
-
 NADPH mutant S199A, pH 7.5, 25°C Clostridium autoethanogenum
1.1.1.4 39000
-
 NADPH wild-type, pH 7.5, 25°C Clostridium autoethanogenum