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Literature summary extracted from

  • Yeon, Y.; Park, H.; Park, K.; Park, H.; Yoo, Y.
    Structural basis for the substrate specificity of 3-hydroxybutyrate dehydrogenase (2016), Biotechnol. Bioprocess Eng., 21, 364-372.
No PubMed abstract available

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.30 expressed in Escherichia coli BL21(DE3) cells Alcaligenes faecalis

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.30 H144A the mutant shows reduced catalytic efficiency with levulinic acid compared to the wild type enzyme Alcaligenes faecalis
1.1.1.30 H144F the mutant shows increased catalytic efficiency with levulinic acid compared to the wild type enzyme Alcaligenes faecalis
1.1.1.30 H144I the mutant shows increased catalytic efficiency with levulinic acid compared to the wild type enzyme Alcaligenes faecalis
1.1.1.30 H144L the mutant shows increased catalytic efficiency with levulinic acid compared to the wild type enzyme Alcaligenes faecalis
1.1.1.30 H144L/W187A the mutant shows reduced catalytic efficiency with levulinic acid compared to the wild type enzyme Alcaligenes faecalis
1.1.1.30 H144L/W187F the mutant shows strongly increased catalytic efficiency with levulinic acid compared to the wild type enzyme Alcaligenes faecalis
1.1.1.30 H144L/W187I the mutant shows reduced catalytic efficiency with levulinic acid compared to the wild type enzyme Alcaligenes faecalis
1.1.1.30 H144L/W187L the mutant shows strongly increased catalytic efficiency with levulinic acid compared to the wild type enzyme Alcaligenes faecalis
1.1.1.30 H144L/W187V the mutant shows reduced catalytic efficiency with levulinic acid compared to the wild type enzyme Alcaligenes faecalis
1.1.1.30 H144V the mutant shows wild type catalytic efficiency with levulinic acid Alcaligenes faecalis
1.1.1.30 W187A the mutant shows reduced catalytic efficiency with levulinic acid compared to the wild type enzyme Alcaligenes faecalis
1.1.1.30 W187F the mutant shows increased catalytic efficiency with levulinic acid compared to the wild type enzyme Alcaligenes faecalis
1.1.1.30 W187I the mutant shows reduced catalytic efficiency with levulinic acid compared to the wild type enzyme Alcaligenes faecalis
1.1.1.30 W187L the mutant shows reduced catalytic efficiency with levulinic acid compared to the wild type enzyme Alcaligenes faecalis
1.1.1.30 W187V the mutant shows reduced catalytic efficiency with levulinic acid compared to the wild type enzyme Alcaligenes faecalis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.30 0.63
-
 levulinic acid mutant enzyme H144L/W187L, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.63
-
 levulinic acid mutant enzyme W187F, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.64
-
 levulinic acid mutant enzyme H144L/W187F, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.78
-
 levulinic acid wild type enzyme, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.91
-
 levulinic acid mutant enzyme H144L, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 1.72
-
 levulinic acid mutant enzyme H144F, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 2.81
-
 levulinic acid mutant enzyme H144I, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 3.84
-
 levulinic acid mutant enzyme H144V, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 4 5.4 levulinic acid mutant enzyme W187I, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 9.87
-
 levulinic acid mutant enzyme H144A, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 13.03
-
 levulinic acid mutant enzyme H144L/W187A, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 13.49
-
 levulinic acid mutant enzyme W187L, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 25.57
-
 levulinic acid mutant enzyme H144L/W187V, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 25.76
-
 levulinic acid mutant enzyme H144L/W187I, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 36.83
-
 levulinic acid mutant enzyme W187A, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 42.1
-
 levulinic acid mutant enzyme W187V, at pH 6.5 and 30°C Alcaligenes faecalis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.30 acetoacetate + NADH + H+ Alcaligenes faecalis
-
 (R)-3-hydroxybutanoate + NAD+
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.30 Alcaligenes faecalis D0VWQ0
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.30 Ni-NTA column chromatography Alcaligenes faecalis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.30 acetoacetate + NADH + H+
-
 Alcaligenes faecalis (R)-3-hydroxybutanoate + NAD+
-
 ?
1.1.1.30 levulinic acid + NADH + H+ the wild type enzyme shows low activity with levulinic acid, while it's a good substrate for mutant enzyme H144L/W187F Alcaligenes faecalis 4-hydroxyvaleric acid + NAD+
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.1.1.30 3HBDH
-
 Alcaligenes faecalis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.30 0.00016
-
 levulinic acid mutant enzyme W187A, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.0003
-
 levulinic acid mutant enzyme W187I, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.0003
-
 levulinic acid mutant enzyme W187V, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.00084
-
 levulinic acid mutant enzyme H144A, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.00122
-
 levulinic acid mutant enzyme H144L/W187A, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.002
-
 levulinic acid mutant enzyme H144L/W187V, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.00289
-
 levulinic acid mutant enzyme H144L/W187I, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.00465
-
 levulinic acid mutant enzyme W187L, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.00766
-
 levulinic acid mutant enzyme H144V, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.00768
-
 levulinic acid wild type enzyme, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.00921
-
 levulinic acid mutant enzyme H144F, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.01069
-
 levulinic acid mutant enzyme H144I, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.02969
-
 levulinic acid mutant enzyme W187F, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.06594
-
 levulinic acid mutant enzyme H144L, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.07107
-
 levulinic acid mutant enzyme H144L/W187L, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.2565
-
 levulinic acid mutant enzyme H144L/W187F, at pH 6.5 and 30°C Alcaligenes faecalis

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.30 NADH
-
 Alcaligenes faecalis

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.1.30 0.006
-
 levulinic acid wild type enzyme, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.006
-
 levulinic acid mutant enzyme W187A, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.008
-
 levulinic acid mutant enzyme H144A, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.013
-
 levulinic acid mutant enzyme W187V, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.014
-
 levulinic acid mutant enzyme W187I, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.016
-
 levulinic acid mutant enzyme H144F, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.016
-
 levulinic acid mutant enzyme H144L/W187A, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.019
-
 levulinic acid mutant enzyme W187F, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.03
-
 levulinic acid mutant enzyme H144I, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.03
-
 levulinic acid mutant enzyme H144V, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.045
-
 levulinic acid mutant enzyme H144L/W187L, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.051
-
 levulinic acid mutant enzyme H144L/W187V, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.06
-
 levulinic acid mutant enzyme H144L, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.063
-
 levulinic acid mutant enzyme W187L, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.074
-
 levulinic acid mutant enzyme H144L/W187I, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.164
-
 levulinic acid mutant enzyme H144L/W187F, at pH 6.5 and 30°C Alcaligenes faecalis