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Literature summary for 1.1.1.76 extracted from

  • Takusagawa, Y.; Otagiri, M.; Ui, S.; Ohtsuki, T.; Mimura, A.; Ohkuma, M.; Kudo, T.
    Purification and characterization of L-2,3-butanediol dehydrogenase of Brevibacterium saccharolyticum C-1012 expressed in Escherichia coli (2001), Biosci. Biotechnol. Biochem., 65, 1876-1878.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
additional information Ba2+, Ca2+, Mn2+, Mg2+ and Co2+ activate enzyme 1.6-1.8 fold at a concentration of 5 mM Corynebacterium glutamicum

Application

Application Comment Organism
synthesis preparation of chiral acetoinic compounds, enzymic identification for chiral acetoinic compounds or as model enzyme for studying the interrelation between enzymic stereospecificity and structure Corynebacterium glutamicum

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli JM109/pLBD2-CTC Corynebacterium glutamicum
L-butanediol gene containing plasmid pLBD2-CTC is expressed in Escherichia coli JM109 Corynebacterium glutamicum

Inhibitors

Inhibitors Comment Organism Structure
2-mercaptoethanol competitive inhibition; competitive inhibitor Corynebacterium glutamicum
acetate slight inhibition at 5 mM Corynebacterium glutamicum
Cu2+ 5 mM, 89% inhibition; 89% inhibition at 5 mM Corynebacterium glutamicum
formate slight inhibition at 5 mM Corynebacterium glutamicum
Hg2+ 100% inhibition at 5 mM; 5 mM, 100% inhibition Corynebacterium glutamicum
Lactate slight inhibition at 5 mM Corynebacterium glutamicum
additional information slight inhibition of 10-20% by organic acids at concentrations of 5 mM: lactate, pyruvate, succinate, acetate and formate Corynebacterium glutamicum
pyruvate slight inhibition at 5 mM Corynebacterium glutamicum
succinate slight inhibition at 5 mM Corynebacterium glutamicum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.07
-
NAD+ pH 6.0 Corynebacterium glutamicum
0.07
-
NAD+ pH 10.5 Corynebacterium glutamicum
0.1
-
NADH pH 6.0 Corynebacterium glutamicum
0.22
-
(S,S)-butane-2,3-diol pH 6.0 Corynebacterium glutamicum
0.22
-
(S,S)-butane-2,3-diol pH 10.5 Corynebacterium glutamicum
0.44
-
L-acetoin pH 6.0 Corynebacterium glutamicum

Metals/Ions

Metals/Ions Comment Organism Structure
Ba2+
-
Corynebacterium glutamicum
Ba2+ Ba2+, Ca2+, Mn2+, Mg2+, and Co2+ activate the enzyme 1.6-1.8fold at a concentration of 5 mM Corynebacterium glutamicum
Ca2+
-
Corynebacterium glutamicum
Ca2+ Ba2+, Ca2+, Mn2+, Mg2+, and Co2+ activate the enzyme 1.6-1.8fold at a concentration of 5 mM Corynebacterium glutamicum
Co2+
-
Corynebacterium glutamicum
Co2+ Ba2+, Ca2+, Mn2+, Mg2+, and Co2+ activate the enzyme 1.6-1.8fold at a concentration of 5 mM Corynebacterium glutamicum
Mg2+
-
Corynebacterium glutamicum
Mg2+ Ba2+, Ca2+, Mn2+, Mg2+, and Co2+ activate the enzyme 1.6-1.8fold at a concentration of 5 mM Corynebacterium glutamicum
Mn2+
-
Corynebacterium glutamicum
Mn2+ Ba2+, Ca2+, Mn2+, Mg2+, and Co2+ activate the enzyme 1.6-1.8fold at a concentration of 5 mM Corynebacterium glutamicum
additional information no significant changes in enzyme activity with 5 mM Ni2+, Zn2+ and Cd2+ Corynebacterium glutamicum

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
27107
-
4 * 30000, SDS-PAGE, 4 * 27107, estimation from gene sequence Corynebacterium glutamicum
30000
-
4 * 30000, SDS-PAGE Corynebacterium glutamicum
30000
-
4 * 30000, SDS-PAGE, 4 * 27107, estimation from gene sequence Corynebacterium glutamicum
110000
-
gel filtration Corynebacterium glutamicum

Organism

Organism UniProt Comment Textmining
Corynebacterium glutamicum
-
C-1012
-
Corynebacterium glutamicum C-1012
-
C-1012
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme Corynebacterium glutamicum
streptomycin sulfate precipitation, affinity chromatography, gel filtration Corynebacterium glutamicum

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
-
Corynebacterium glutamicum
277.6
-
-
Corynebacterium glutamicum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2,3-pentanedione + NADH + H+ 7% activity in comparison to L-acetoin Corynebacterium glutamicum ?
-
r
2,3-pentanedione + NADH + H+ 7% activity in comparison to L-acetoin Corynebacterium glutamicum C-1012 ?
-
r
diacetyl + NADH also reduction of 2,3-pentanedione Corynebacterium glutamicum L-acetoin + NAD+
-
?
diacetyl + NADH also reduction of 2,3-pentanedione Corynebacterium glutamicum C-1012 L-acetoin + NAD+
-
?
diacetyl + NADH + H+ 35% activity in comparison to L-acetoin Corynebacterium glutamicum ?
-
r
L-acetoin + NADH short chain dehydrogenase reductase family Corynebacterium glutamicum L-2,3-butanediol
-
r
L-acetoin + NADH no oxidadion of several alcohols Corynebacterium glutamicum L-2,3-butanediol
-
r
L-acetoin + NADH short chain dehydrogenase reductase family Corynebacterium glutamicum C-1012 L-2,3-butanediol
-
r
L-acetoin + NADH no oxidadion of several alcohols Corynebacterium glutamicum C-1012 L-2,3-butanediol
-
r
L-acetoin + NADH + H+ 100% activity Corynebacterium glutamicum (S,S)-butane-2,3-diol + NAD+
-
r
additional information not: meso-butanediol, D-butanediol, 2-butanol, 1,2-propanediol, ethanol, acetol, 1,2-butanediol, 1,3-butanediol, n-butanol, n-propanol, D-acetoin, acetol, dihydroxyacetone, 2,4-pentanedione Corynebacterium glutamicum ?
-
?
additional information not: meso-butanediol, D-butanediol, 2-butanol, 1,2-propanediol, ethanol, acetol, 1,2-butanediol, 1,3-butanediol, n-butanol, n-propanol, D-acetoin, acetol, dihydroxyacetone, 2,4-pentanedione Corynebacterium glutamicum C-1012 ?
-
?

Subunits

Subunits Comment Organism
tetramer 4 * 30000, SDS-PAGE Corynebacterium glutamicum
tetramer 4 * 30000, SDS-PAGE, 4 * 27107, estimation from gene sequence Corynebacterium glutamicum

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
37
-
up to, rapidly inactivated over 40°C, 30 min incubation before the enzyme assay Corynebacterium glutamicum
37 40 The enzyme is stable up to 37°C but rapidly inactivated over 40°C Corynebacterium glutamicum

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6
-
reduction of L-acetoin Corynebacterium glutamicum
10.5
-
oxidation of L-butanediol Corynebacterium glutamicum
10.5
-
oxidation of (S,S)-butane-2,3-diol Corynebacterium glutamicum

pH Stability

pH Stability pH Stability Maximum Comment Organism
7 9
-
Corynebacterium glutamicum
7 9 stable Corynebacterium glutamicum

Cofactor

Cofactor Comment Organism Structure
NAD+
-
Corynebacterium glutamicum
NADH
-
Corynebacterium glutamicum

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
4.28
-
2-mercaptoethanol Dixon plot Corynebacterium glutamicum
4.28
-
2-mercaptoethanol pH 6.0 Corynebacterium glutamicum

pI Value

Organism Comment pI Value Maximum pI Value
Corynebacterium glutamicum isoelectric focussing
-
4.9