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Literature summary extracted from

  • Lee, P.; Raj, S.; Zhou, S.; Ashok, S.; Edwardraja, S.; Park, S.
    3-Hydroxyisobutyrate dehydrogenase-I from Pseudomonas denitrificans ATCC 13867 degrades 3-hydroxypropionic acid (2014), Biotechnol. Bioprocess Eng., 19, 1-7.
No PubMed abstract available

Activating Compound

EC Number Activating Compound Comment Organism Structure
1.1.1.31 2-mercaptoethanol about 1575.34% activity at 1 mM Pseudomonas denitrificans (nom. rej.)
1.1.1.31 dithiothreitol about 182.11% activity at 1 mM Pseudomonas denitrificans (nom. rej.)
1.1.1.31 EDTA about 156.21% activity at 1 mM Pseudomonas denitrificans (nom. rej.)

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.31 expressed in Escherichia coli BL21(DE3) cells Pseudomonas denitrificans (nom. rej.)

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.1.31 Ag+ complete inhibition at 1 mM Pseudomonas denitrificans (nom. rej.)
1.1.1.31 Co2+ about 68.1% residual activity at 1 mM Pseudomonas denitrificans (nom. rej.)
1.1.1.31 Cu2+ about 15.67% residual activity at 1 mM Pseudomonas denitrificans (nom. rej.)
1.1.1.31 Fe2+ complete inhibition at 1 mM Pseudomonas denitrificans (nom. rej.)
1.1.1.31 Hg2+ complete inhibition at 1 mM Pseudomonas denitrificans (nom. rej.)
1.1.1.31 K+ about 92.11% residual activity at 1 mM Pseudomonas denitrificans (nom. rej.)
1.1.1.31 Li+ about 92.32% residual activity at 1 mM Pseudomonas denitrificans (nom. rej.)
1.1.1.31 Mg2+ about 87.34% residual activity at 1 mM Pseudomonas denitrificans (nom. rej.)
1.1.1.31 NH4+ about 90.17% residual activity at 1 mM Pseudomonas denitrificans (nom. rej.)
1.1.1.31 Ni2+ about 63.67% residual activity at 1 mM Pseudomonas denitrificans (nom. rej.)
1.1.1.31 Zn2+ about 16.13% residual activity at 1 mM Pseudomonas denitrificans (nom. rej.)

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.31 0.12
-
 (S)-3-hydroxyisobutyrate at 37°C and pH 9.0 Pseudomonas denitrificans (nom. rej.)
1.1.1.31 2.09
-
 3-hydroxypropionic acid at 37°C and pH 9.0 Pseudomonas denitrificans (nom. rej.)
1.1.1.31 10.37
-
 methyl DL-serine at 37°C and pH 9.0 Pseudomonas denitrificans (nom. rej.)
1.1.1.31 10.48
-
 methyl (2S)-3-hydroxy-2-methylpropanoate at 37°C and pH 9.0 Pseudomonas denitrificans (nom. rej.)
1.1.1.31 31.84
-
 L-serine at 37°C and pH 9.0 Pseudomonas denitrificans (nom. rej.)

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.1.1.31 Mn2+ about 127.2% activity at 1 mM Pseudomonas denitrificans (nom. rej.)
1.1.1.31 additional information not influenced by Na+ Pseudomonas denitrificans (nom. rej.)

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.31 25000
-
 x * 25000, SDS-PAGE Pseudomonas denitrificans (nom. rej.)

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.31 Pseudomonas denitrificans (nom. rej.)
-
-
-
1.1.1.31 Pseudomonas denitrificans (nom. rej.) ATCC 13867
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.31 Ni-NTA column chromatography and Superdex 200 gel filtration Pseudomonas denitrificans (nom. rej.)

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.1.1.31 1.62
-
 crude extract, at 37°C and pH 9.0 Pseudomonas denitrificans (nom. rej.)
1.1.1.31 58.67
-
 after 36.22fold purification, at 37°C and pH 9.0 Pseudomonas denitrificans (nom. rej.)

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.31 (S)-3-hydroxyisobutyrate + NAD+ the enzyme is very specific to (S)-3-hydroxyisobutyrate Pseudomonas denitrificans (nom. rej.) 2-methyl-3-oxopropanoate + NADH + H+
-
 ?
1.1.1.31 (S)-3-hydroxyisobutyrate + NAD+ the enzyme is very specific to (S)-3-hydroxyisobutyrate Pseudomonas denitrificans (nom. rej.) ATCC 13867 2-methyl-3-oxopropanoate + NADH + H+
-
 ?
1.1.1.31 3-hydroxypropionic acid + NAD+
-
 Pseudomonas denitrificans (nom. rej.) malonate semialdehyde + NADH + H+
-
 r
1.1.1.31 3-hydroxypropionic acid + NAD+
-
 Pseudomonas denitrificans (nom. rej.) ATCC 13867 malonate semialdehyde + NADH + H+
-
 r
1.1.1.31 L-serine + NAD+
-
 Pseudomonas denitrificans (nom. rej.) 3-oxo-L-alanine + NADH + H+
-
 r
1.1.1.31 L-serine + NAD+
-
 Pseudomonas denitrificans (nom. rej.) ATCC 13867 3-oxo-L-alanine + NADH + H+
-
 r
1.1.1.31 methyl (2S)-3-hydroxy-2-methylpropanoate + NAD+
-
 Pseudomonas denitrificans (nom. rej.) methyl (2S)-2-methyl-3-oxopropanoate + NADH + H+
-
 r
1.1.1.31 methyl (2S)-3-hydroxy-2-methylpropanoate + NAD+
-
 Pseudomonas denitrificans (nom. rej.) ATCC 13867 methyl (2S)-2-methyl-3-oxopropanoate + NADH + H+
-
 r
1.1.1.31 methyl DL-serine + NAD+
-
 Pseudomonas denitrificans (nom. rej.) methyl 3-oxo-DL-alanine + NADH + H+
-
 r
1.1.1.31 additional information no activity with (R)-3-hydroxyisobutyrate Pseudomonas denitrificans (nom. rej.) ?
-
 ?
1.1.1.31 additional information no activity with (R)-3-hydroxyisobutyrate Pseudomonas denitrificans (nom. rej.) ATCC 13867 ?
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.1.31 ? x * 25000, SDS-PAGE Pseudomonas denitrificans (nom. rej.)

Synonyms

EC Number Synonyms Comment Organism
1.1.1.31 3-hydroxyisobutyrate dehydrogenase-I
-
 Pseudomonas denitrificans (nom. rej.)
1.1.1.31 3HIBDHI
-
 Pseudomonas denitrificans (nom. rej.)

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.31 37
-
-
 Pseudomonas denitrificans (nom. rej.)

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
1.1.1.31 30 45 more than 50% activity between 30 and 40°C. Approximately 80% of the activity is lost at 45°C Pseudomonas denitrificans (nom. rej.)

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.31 7.94
-
 methyl DL-serine at 37°C and pH 9.0 Pseudomonas denitrificans (nom. rej.)
1.1.1.31 32.17
-
 3-hydroxypropionic acid at 37°C and pH 9.0 Pseudomonas denitrificans (nom. rej.)
1.1.1.31 41.35
-
 methyl (2S)-3-hydroxy-2-methylpropanoate at 37°C and pH 9.0 Pseudomonas denitrificans (nom. rej.)
1.1.1.31 47.16
-
 L-serine at 37°C and pH 9.0 Pseudomonas denitrificans (nom. rej.)
1.1.1.31 70.81
-
 (S)-3-hydroxyisobutyrate at 37°C and pH 9.0 Pseudomonas denitrificans (nom. rej.)

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.31 9
-
-
 Pseudomonas denitrificans (nom. rej.)

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.1.1.31 8 10 approximately 40% of the activity is retained at pH 11.0. At pH 7.0, the activity is reduced to 30% of the activity observed at pH 9.0 Pseudomonas denitrificans (nom. rej.)

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.31 NAD+ the enzyme prefers NAD+ over NADP+ as a cofactor Pseudomonas denitrificans (nom. rej.)

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.1.31 0.75
-
 methyl DL-serine at 37°C and pH 9.0 Pseudomonas denitrificans (nom. rej.)
1.1.1.31 1.46
-
 L-serine at 37°C and pH 9.0 Pseudomonas denitrificans (nom. rej.)
1.1.1.31 4.04
-
 methyl (2S)-3-hydroxy-2-methylpropanoate at 37°C and pH 9.0 Pseudomonas denitrificans (nom. rej.)
1.1.1.31 15.4
-
 3-hydroxypropionic acid at 37°C and pH 9.0 Pseudomonas denitrificans (nom. rej.)
1.1.1.31 604.1
-
 (S)-3-hydroxyisobutyrate at 37°C and pH 9.0 Pseudomonas denitrificans (nom. rej.)