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Literature summary for 1.1.1.31 extracted from

  • Yao, T.; Xu, L.; Ying, H.; Huang, H.; Yan, M.
    The catalytic property of 3-hydroxyisobutyrate dehydrogenase from Bacillus cereus on 3-hydroxypropionate (2009), Appl. Biochem. Biotechnol., 160, 694-703.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
additional information ethylene glycol tetraacetic acid or EDTA (all at 0.005 and 0.2 mM) do not affect 3-HIBADH activity Bacillus cereus

Application

Application Comment Organism
additional information 3-HIBADH may play a role in biosynthesis of 3-hydroxypropionate as a biological source Bacillus cereus

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21 (DE3) Bacillus cereus
MmsB gene, overexpression in Escherichia coli strain BL21, subcloning in strain DH5alpha Bacillus cereus
pEB81 recombinant vector introduced into Escherichia coli BL21 to overexpress 3-HIBADH protein Bacillus cereus

Inhibitors

Inhibitors Comment Organism Structure
additional information is not inhibited by 1-propanol, lactate potassium, malate potassium, malonate potassium, acetaldehyde, ethanol, and glycerin Bacillus cereus
Zn2+ inhibits at 0.2 mM Bacillus cereus
ZnCl2 0.2 mM, 60% inhibition; inhibits approximately 60% of 3-HIBADH activity at 0.2 mM but shows no effect at 0.005 mM Bacillus cereus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.25
-
NADP+
-
Bacillus cereus
0.25
-
NADP+ pH 8.5, 37┬░C Bacillus cereus
2.4
-
NAD+
-
Bacillus cereus
2.4
-
NAD+ pH 8.5, 37┬░C Bacillus cereus
16.8
-
3-hydroxypropionate
-
Bacillus cereus
16.8
-
3-hydroxypropionate pH 8.5, 37┬░C, KM-value for 3-hydroxyisobutyrate is approximately 20fold lower Bacillus cereus

Metals/Ions

Metals/Ions Comment Organism Structure
additional information no metal ion requirement Bacillus cereus
additional information has no metal ion requirement, MnSO4, CuSO4, CaCl2, MgSO4, and FeSO4 have no effect Bacillus cereus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
32000
-
gel filtration Bacillus cereus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3-hydroxy-2-methylpropanoate + NAD+ Bacillus cereus
-
2-methyl-3-oxopropanoate + NADH + H+
-
r
3-hydroxy-2-methylpropanoate + NAD+ Bacillus cereus ATCC 14579
-
2-methyl-3-oxopropanoate + NADH + H+
-
r
3-hydroxy-2-methylpropanoate + NADP+ Bacillus cereus
-
2-methyl-3-oxopropanoate + NADPH + H+
-
r
3-hydroxy-2-methylpropanoate + NADP+ Bacillus cereus ATCC 14579
-
2-methyl-3-oxopropanoate + NADPH + H+
-
r
additional information Bacillus cereus MmsB from Bacillus cereus exhibits 3-hydroxyisobutyrate dehydrogenase as well as 3-hydroxypropionate dehydrogenase activity, EC 1.1.1.59 ?
-
?
additional information Bacillus cereus ATCC 14579 MmsB from Bacillus cereus exhibits 3-hydroxyisobutyrate dehydrogenase as well as 3-hydroxypropionate dehydrogenase activity, EC 1.1.1.59 ?
-
?

Organism

Organism UniProt Comment Textmining
Bacillus cereus
-
ATCC 14579
-
Bacillus cereus
-
-
-
Bacillus cereus
-
MmsB gene
-
Bacillus cereus ATCC 14579
-
-
-
Bacillus cereus ATCC 14579
-
MmsB gene
-

Purification (Commentary)

Purification (Comment) Organism
96fold purified at 4┬░C by ammonium sulfate fractionation and gel filtration Bacillus cereus
ammonium sulfate precipitation, hydrophobic interaction chromatography (Phenyl-Sepharose), anion exchange chromatography Bacillus cereus
native enzyme 100fold by ammonium sulfate fractionation, and hydrophobic interaction and anion exchange chromatography Bacillus cereus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [┬Ámol/min/mg] Specific Activity Maximum [┬Ámol/min/mg] Comment Organism
0.091
-
homogenate Bacillus cereus
8.7
-
purified enzyme Bacillus cereus
8.77
-
pH 8.5, 37┬░C Bacillus cereus
8.77
-
purified enzyme, pH 8.5, 37┬░C Bacillus cereus

Storage Stability

Storage Stability Organism
-20┬░C, 50 mM Tris-HCl buffer, pH 8.5, 2 mM EDTA, 2 mM DTT, 50% glycerol Bacillus cereus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3-hydroxy-2-methylpropanoate + NAD+
-
Bacillus cereus 2-methyl-3-oxopropanoate + NADH + H+
-
r
3-hydroxy-2-methylpropanoate + NAD+
-
Bacillus cereus ATCC 14579 2-methyl-3-oxopropanoate + NADH + H+
-
r
3-hydroxy-2-methylpropanoate + NADP+
-
Bacillus cereus 2-methyl-3-oxopropanoate + NADPH + H+
-
r
3-hydroxy-2-methylpropanoate + NADP+
-
Bacillus cereus ATCC 14579 2-methyl-3-oxopropanoate + NADPH + H+
-
r
3-hydroxyisobutyrate + NAD+
-
Bacillus cereus methylmalonate semialdehyde + NADH + H+
-
r
3-hydroxypropanoate + NAD(P)+ is more active with NADP+ than NAD+ Bacillus cereus ?
-
?
3-hydroxypropanoate + NAD(P)+ is more active with NADP+ than NAD+ Bacillus cereus ATCC 14579 ?
-
?
3-hydroxypropanoate + NAD+
-
Bacillus cereus malonate semialdehyde + NADH + H+
-
r
3-hydroxypropanoate + NADP+
-
Bacillus cereus malonate semialdehyde + NADPH + H+
-
r
additional information MmsB from Bacillus cereus exhibits 3-hydroxyisobutyrate dehydrogenase as well as 3-hydroxypropionate dehydrogenase activity, EC 1.1.1.59 Bacillus cereus ?
-
?
additional information wide substrate specificity of the 3-HIBADH Bacillus cereus ?
-
?
additional information MmsB from Bacillus cereus exhibits 3-hydroxyisobutyrate dehydrogenase as well as 3-hydroxypropionate dehydrogenase activity, EC 1.1.1.59 Bacillus cereus ATCC 14579 ?
-
?
additional information wide substrate specificity of the 3-HIBADH Bacillus cereus ATCC 14579 ?
-
?

Subunits

Subunits Comment Organism
? x * 32000, SDS-PAGE Bacillus cereus

Synonyms

Synonyms Comment Organism
3-HIBADH
-
Bacillus cereus
3-hydroxyisobutyrate dehydrogenase cf. EC1.1.1.59, wide substrate specificity (hydroxyacid derivatives) Bacillus cereus

Temperature Optimum [┬░C]

Temperature Optimum [┬░C] Temperature Optimum Maximum [┬░C] Comment Organism
37
-
-
Bacillus cereus
37
-
assay at Bacillus cereus

Temperature Range [┬░C]

Temperature Minimum [┬░C] Temperature Maximum [┬░C] Comment Organism
35 45 shows more than 90% of its optimized activity Bacillus cereus

Temperature Stability [┬░C]

Temperature Stability Minimum [┬░C] Temperature Stability Maximum [┬░C] Comment Organism
45
-
enzyme activity is not decreased during preincubation at 45┬░C within 30 min, but preincubation at 55┬░C in 3 min can significantly denaturate and deactivate the enzyme Bacillus cereus
45
-
stable at 45┬░C for 30 min, about 40% activity after 3 min at 55┬░C Bacillus cereus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.21
-
3-hydroxypropionate
-
Bacillus cereus
0.21
-
3-hydroxypropionate pH 8.5, 37┬░C Bacillus cereus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.5
-
assay at Bacillus cereus
8.8 9
-
Bacillus cereus
8.8 9 oxidation of 3-hydroxypropionate by 3-HIBADH Bacillus cereus

pH Range

pH Minimum pH Maximum Comment Organism
7 10
-
Bacillus cereus
7 10 oxidation of 3-hydroxypropionate by 3-HIBADH Bacillus cereus

Cofactor

Cofactor Comment Organism Structure
additional information 3-HIBADH is more active with NADP+ than NAD+ Bacillus cereus
NAD(P)+ is more active with NADP+ than NAD+ Bacillus cereus
NAD+
-
Bacillus cereus
NADH
-
Bacillus cereus
NADP+
-
Bacillus cereus
NADPH
-
Bacillus cereus

General Information

General Information Comment Organism
physiological function 3-hydroxyisobutyrate dehydrogenase is a key enzyme for the metabolism of valine and some keto-bodies. It can catalyze reversible conversion of 3-hydroxyisobutyrate to methylmalonate semialdehyde Bacillus cereus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.0125
-
3-hydroxypropionate pH 8.5, 37┬░C Bacillus cereus