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Literature summary for 1.1.1.31 extracted from

  • Srikalaivani, R.; Singh, A.; Vijayan, M.; Surolia, A.
    Structure, interactions and action of Mycobacterium tuberculosis 3-hydroxyisobutyric acid dehydrogenase (2018), Biochem. J., 475, 2457-2471 .
    View publication on PubMed
Search for more literature for 1.1.1.31

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(lambdaDE3) cells Mycobacterium tuberculosis

Crystallization (Commentary)

Crystallization (Comment) Organism
native enzyme, the holoenzyme, binary complexes with NAD+, (S)-3-hydroxyisobutyrate, (R)-3-hydroxyisobutyrate, L-serine and 3-hydroxypropanoic acid and ternary complexes involving the substrates and NAD+, microbatch-under-oil method, using 0.02 M MgCl2, 0.1 M HEPES (pH 7.5), 22% (w/v) polyacrylic acid sodium salt 5100 Mycobacterium tuberculosis

Protein Variants

Protein Variants Comment Organism
D244A the mutant shows strong reduction in the activity compared to the wild type enzyme Mycobacterium tuberculosis
D275A the mutant shows a marginal loss of activity compared to the wild type enzyme Mycobacterium tuberculosis
F236A the mutant shows reduction in the activity compared to the wild type enzyme Mycobacterium tuberculosis
K168A inactive Mycobacterium tuberculosis
N172A the mutant shows dramatic reduction in the activity compared to the wild type enzyme Mycobacterium tuberculosis
S119A inactive Mycobacterium tuberculosis
T207A the mutant shows reduction in the activity compared to the wild type enzyme Mycobacterium tuberculosis
W211A the mutant shows dramatic reduction in the activity compared to the wild type enzyme Mycobacterium tuberculosis

Inhibitors

Inhibitors Comment Organism Structure
NADH competitive inhibition in the presence of NADH when NAD+ is varied, noncompetitive inhibition with NADH when (S)-3-hydroxyisobutyrate is varied Mycobacterium tuberculosis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.563
-
 NAD+ with (S)-3-hydroxyisobutyrate as cosubstrate, pH 8.5 and 30°C Mycobacterium tuberculosis
0.574
-
 (S)-3-hydroxyisobutyrate at pH 8.5 and 30°C Mycobacterium tuberculosis
0.582
-
 NADP+ with (S)-3-hydroxyisobutyrate as cosubstrate, pH 8.5 and 30°C Mycobacterium tuberculosis
2.5 3 3-hydroxypropanoic acid at pH 8.5 and 30°C Mycobacterium tuberculosis
4.56
-
 (R)-3-hydroxyisobutyrate at pH 8.5 and 30°C Mycobacterium tuberculosis
21.33
-
 L-serine at pH 8.5 and 30°C Mycobacterium tuberculosis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
66000
-
 gel filtration Mycobacterium tuberculosis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
(S)-3-hydroxyisobutyrate + NAD+ Mycobacterium tuberculosis the enzyme is specific to (S)-3-hydroxyisobutyrate and prefers NAD+ as cofactor 2-methyl-3-oxopropanoate + NADH + H+
-
 ?
(S)-3-hydroxyisobutyrate + NAD+ Mycobacterium tuberculosis H37Rv the enzyme is specific to (S)-3-hydroxyisobutyrate and prefers NAD+ as cofactor 2-methyl-3-oxopropanoate + NADH + H+
-
 ?

Organism

Organism UniProt Comment Textmining
Mycobacterium tuberculosis P9WNY5
-
-
Mycobacterium tuberculosis H37Rv P9WNY5
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA resin column chromatography Mycobacterium tuberculosis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(R)-3-hydroxyisobutyrate + NAD+ substrate with much less efficiency compared to (S)-3-hydroxyisobutyrate Mycobacterium tuberculosis 2-methyl-3-oxopropanoate + NADH + H+
-
 ?
(R)-3-hydroxyisobutyrate + NAD+ substrate with much less efficiency compared to (S)-3-hydroxyisobutyrate Mycobacterium tuberculosis H37Rv 2-methyl-3-oxopropanoate + NADH + H+
-
 ?
(S)-3-hydroxyisobutyrate + NAD+ the enzyme is specific to (S)-3-hydroxyisobutyrate and prefers NAD+ as cofactor Mycobacterium tuberculosis 2-methyl-3-oxopropanoate + NADH + H+
-
 ?
(S)-3-hydroxyisobutyrate + NAD+ the enzyme is specific to (S)-3-hydroxyisobutyrate Mycobacterium tuberculosis 2-methyl-3-oxopropanoate + NADH + H+
-
 ?
(S)-3-hydroxyisobutyrate + NAD+ the enzyme is specific to (S)-3-hydroxyisobutyrate and prefers NAD+ as cofactor Mycobacterium tuberculosis H37Rv 2-methyl-3-oxopropanoate + NADH + H+
-
 ?
(S)-3-hydroxyisobutyrate + NAD+ the enzyme is specific to (S)-3-hydroxyisobutyrate Mycobacterium tuberculosis H37Rv 2-methyl-3-oxopropanoate + NADH + H+
-
 ?
(S)-3-hydroxyisobutyrate + NADP+
-
 Mycobacterium tuberculosis 2-methyl-3-oxopropanoate + NADPH + H+
-
 ?
3-hydroxypropanoic acid + NAD+ substrate with much less efficiency compared to (S)-3-hydroxyisobutyrate Mycobacterium tuberculosis 3-oxopropanoate + NADH + H+
-
 ?
L-serine + NAD+ substrate with much less efficiency compared to (S)-3-hydroxyisobutyrate Mycobacterium tuberculosis ?
-
 ?
L-serine + NAD+ substrate with much less efficiency compared to (S)-3-hydroxyisobutyrate Mycobacterium tuberculosis H37Rv ?
-
 ?
additional information no activity with D-serine, (S)-3-hydroxyisobutyrate methyl ester, (R)-3-hydroxyisobutyrate methyl ester, L-threonine and D-threonine Mycobacterium tuberculosis ?
-
-
additional information no activity with D-serine, (S)-3-hydroxyisobutyrate methyl ester, (R)-3-hydroxyisobutyrate methyl ester, L-threonine and D-threonine Mycobacterium tuberculosis H37Rv ?
-
-

Subunits

Subunits Comment Organism
homodimer 2 * 30875, calculated from amino acid sequence Mycobacterium tuberculosis

Synonyms

Synonyms Comment Organism
3-hydroxyisobutyric acid dehydrogenase
-
 Mycobacterium tuberculosis
HIBADH
-
 Mycobacterium tuberculosis

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
34.5
-
 at pH 11.0, the melting temperature is 34.5°C Mycobacterium tuberculosis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.205
-
 (R)-3-hydroxyisobutyrate at pH 8.5 and 30°C Mycobacterium tuberculosis
0.262
-
 L-serine at pH 8.5 and 30°C Mycobacterium tuberculosis
0.29
-
 NADP+ with (S)-3-hydroxyisobutyrate as cosubstrate, pH 8.5 and 30°C Mycobacterium tuberculosis
1.018
-
 (S)-3-hydroxyisobutyrate at pH 8.5 and 30°C Mycobacterium tuberculosis
1.182
-
 NAD+ with (S)-3-hydroxyisobutyrate as cosubstrate, pH 8.5 and 30°C Mycobacterium tuberculosis
2.53
-
 3-hydroxypropanoic acid at pH 8.5 and 30°C Mycobacterium tuberculosis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
11
-
-
 Mycobacterium tuberculosis

Cofactor

Cofactor Comment Organism Structure
NAD+ the enzyme preferentially uses NAD+ as the cofactor Mycobacterium tuberculosis

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.044
-
 L-serine at pH 8.5 and 30°C Mycobacterium tuberculosis
0.0448
-
 (R)-3-hydroxyisobutyrate at pH 8.5 and 30°C Mycobacterium tuberculosis
0.1036
-
 3-hydroxypropanoic acid at pH 8.5 and 30°C Mycobacterium tuberculosis
0.498
-
 NADP+ with (S)-3-hydroxyisobutyrate as cosubstrate, pH 8.5 and 30°C Mycobacterium tuberculosis
1.773
-
 (S)-3-hydroxyisobutyrate at pH 8.5 and 30°C Mycobacterium tuberculosis
2.1
-
 NAD+ with (S)-3-hydroxyisobutyrate as cosubstrate, pH 8.5 and 30°C Mycobacterium tuberculosis