BRENDA - Enzyme Database
show all sequences of 1.1.1.337

L-2-Hydroxyisocaproate dehydrogenase - A new enzyme from Lactobacillus confusus for the stereospecific reduction of 2-ketocarboxylic acids

Schütte, W.; Hummel, W.; Kula, M.R.; Appl. Microbiol. Biotechnol. 19, 167-176 (1984)
No PubMed abstract available

Data extracted from this reference:

Inhibitors
Inhibitors
Commentary
Organism
Structure
1,10-phenanthroline
10 mM, 15% inhibition
Weissella confusa
4-chloromercuribenzoate
weak inhibition
Weissella confusa
iodoacetamide
weak inhibition
Weissella confusa
iodoacetate
weak inhibition
Weissella confusa
KCN
weak inhibition
Weissella confusa
additional information
reduced glutathione (20 mM) has no effect. EDTA, 2,2'-bipyridyl in 1 mM and 10 mM final concentrations have no effect on enzymatic activity
Weissella confusa
NADH
inhibition above 0.24 mM
Weissella confusa
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.06
-
2-oxoisocaproate
pH 7.0, 30°C
Weissella confusa
0.065
-
2-oxoisovalerate
pH 7.0, 30°C
Weissella confusa
0.1
-
2-oxovalerate
pH 7.0, 30°C
Weissella confusa
0.1
2
phenylpyruvate
pH 7.0, 30°C
Weissella confusa
0.1
2
2-oxocaproate
pH 7.0, 30°C
Weissella confusa
0.17
-
2-oxooctanoate
pH 7.0, 30°C
Weissella confusa
0.45
-
2-oxobutyrate
pH 7.0, 30°C
Weissella confusa
0.6
-
2-Hydroxyisovalerate
pH 8.0, 30°C
Weissella confusa
0.62
-
2-hydroxyisocaproate
pH 8.0, 30°C
Weissella confusa
0.64
-
3-Phenyllactate
pH 8.0, 30°C
Weissella confusa
1.8
-
2-hydroxyvalerate
pH 8.0, 30°C
Weissella confusa
1.9
-
2-Hydroxyoctanoate
pH 8.0, 30°C
Weissella confusa
2.2
-
2-hydroxycaproate
pH 8.0, 30°C
Weissella confusa
4.5
-
pyruvate
pH 7.0, 30°C
Weissella confusa
100
-
L-lactate
pH 8.0, 30°C
Weissella confusa
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
additional information
at 1 mM no influence is observed with Mg2+, Ca2+, Co2+, Ni2+, and Cd2+ and very weak effects with Mn2+, Cu2+, and Zn2+
Weissella confusa
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
33000
-
4 * 33000, SDS-PAGE
Weissella confusa
125000
-
gel filtration
Weissella confusa
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
a 2-oxocarboxylate + NADH + H+
Weissella confusa
-
(2S)-2-hydroxycarboxylate + NAD+
-
-
r
Organism
Organism
UniProt
Commentary
Textmining
Weissella confusa
P14295
i.e. Lactobacillus confusus
-
Purification (Commentary)
Purification (Commentary)
Organism
-
Weissella confusa
Source Tissue
Source Tissue
Commentary
Organism
Textmining
culture condition
maximum specific and volume activities are found at the end of the exponential growth phase, during further fermentation enzyme activity declines rapidly and is no longer detectable after 15 h
Weissella confusa
-
Storage Stability
Storage Stability
Organism
4°C, pH 6.5, concentrated enzyme solution, 10% loss of activity after 6 months
Weissella confusa
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
2-hydroxycaproate + NAD+
-
726750
Weissella confusa
2-oxocaproate + NADH + H+
-
-
-
r
2-hydroxyisocaproate + NAD+
the initial rate of the reduction of 2-oxoisocaproate is about seven times faster than the reverse reaction, the dehydrogenation of L-2-hydroxyisocaproate, measured in buffers at the respective pH optima
726750
Weissella confusa
2-oxoisocaproate + NADH + H+
-
-
-
r
2-hydroxyisovalerate + NAD+
-
726750
Weissella confusa
2-oxoisovalerate + NADH + H+
-
-
-
r
2-hydroxyoctanoate + NAD+
-
726750
Weissella confusa
2-oxooctanoate + NADH + H+
-
-
-
?
2-hydroxyvalerate + NAD+
-
726750
Weissella confusa
2-oxovalerate + NADH + H+
-
-
-
r
2-oxobutyrate + NADH + H+
-
726750
Weissella confusa
2-hydroxybutyrate + NAD+
-
-
-
r
2-oxocaproate + NADH + H+
-
726750
Weissella confusa
2-hydroxycaproate + NAD+
-
-
-
r
2-oxoisocaproate + NADH + H+
2-oxoisocaproate is the best substrate with the lowest KM-valus of 0.065 mM. Tthe initial rate of the reduction of 2-oxoisocaproate is about seven times faster than the reverse reaction, the dehydrogenation of L-2-hydroxyisocaproate, measured in buffers at the respective pH optima
726750
Weissella confusa
2-hydroxyisocaproate + NAD+
-
-
-
r
2-oxoisovalerate + NADH + H+
-
726750
Weissella confusa
2-hydroxyisovalerate + NAD+
-
-
-
r
2-oxooctanoate + NADH + H+
-
726750
Weissella confusa
2-hydroxyoctanoate + NAD+
-
-
-
r
2-oxovalerate + NADH + H+
-
726750
Weissella confusa
2-hydroxyvalerate + NAD+
-
-
-
r
3-phenyllactate + NAD+
-
726750
Weissella confusa
pyruvate + NADH + H+
-
-
-
r
a 2-oxocarboxylate + NADH + H+
-
726750
Weissella confusa
(2S)-2-hydroxycarboxylate + NAD+
-
-
-
r
a 2-oxocarboxylate + NADH + H+
various 2-oxocarboxylic acids are stereospecifically reduced to the corresponding (S)-2-hydroxycarboxylic acids. In the reverse reaction the NAD+-dependent dehydrogenation of L-2-hydroxycarboxylic acids is observed
726750
Weissella confusa
(2S)-2-hydroxycarboxylate + NAD+
-
-
-
r
L-lactate + NAD+
-
726750
Weissella confusa
pyruvate + NADH + H+
-
-
-
r
phenylpyruvate + NADH + H+
-
726750
Weissella confusa
3-phenyllactate + NAD+
-
-
-
r
pyruvate + NADH + H+
-
726750
Weissella confusa
L-lactate + NAD+
-
-
-
r
Subunits
Subunits
Commentary
Organism
tetramer
4 * 33000, SDS-PAGE
Weissella confusa
Synonyms
Synonyms
Commentary
Organism
L-2-hydroxyisocaproate dehydrogenase
-
Weissella confusa
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
30
-
assay at
Weissella confusa
50
-
-
Weissella confusa
Temperature Range [°C]
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
Organism
30
50
the maximal temperature is around 50°C. At this temperature the reaction proceeded at about 3fold increase of the rate compared to 30°C
Weissella confusa
Temperature Stability [°C]
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
40
-
1 h, stable up to
Weissella confusa
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7
-
reduction of 2-oxoisocaproate
Weissella confusa
8
8.5
dehydrogenation of L-2-hydroxyisocaproate
Weissella confusa
pH Stability
pH Stability
pH Stability Maximum
Commentary
Organism
5.5
9
4°C, 24 h, stable
Weissella confusa
6.5
-
4°C, 6 months, concentrated enzyme solutions, 10% loss of activity
Weissella confusa
Cofactor
Cofactor
Commentary
Organism
Structure
NAD+
the enzyme requires NAD+ as a cofactor, no reaction with NADP+ in concentrations up to 3 mM
Weissella confusa
NADH
the enzyme requires NADH as a cofactor, no reaction with NADPH up to 0.24 mM
Weissella confusa
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NAD+
the enzyme requires NAD+ as a cofactor, no reaction with NADP+ in concentrations up to 3 mM
Weissella confusa
NADH
the enzyme requires NADH as a cofactor, no reaction with NADPH up to 0.24 mM
Weissella confusa
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
1,10-phenanthroline
10 mM, 15% inhibition
Weissella confusa
4-chloromercuribenzoate
weak inhibition
Weissella confusa
iodoacetamide
weak inhibition
Weissella confusa
iodoacetate
weak inhibition
Weissella confusa
KCN
weak inhibition
Weissella confusa
additional information
reduced glutathione (20 mM) has no effect. EDTA, 2,2'-bipyridyl in 1 mM and 10 mM final concentrations have no effect on enzymatic activity
Weissella confusa
NADH
inhibition above 0.24 mM
Weissella confusa
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.06
-
2-oxoisocaproate
pH 7.0, 30°C
Weissella confusa
0.065
-
2-oxoisovalerate
pH 7.0, 30°C
Weissella confusa
0.1
-
2-oxovalerate
pH 7.0, 30°C
Weissella confusa
0.1
2
phenylpyruvate
pH 7.0, 30°C
Weissella confusa
0.1
2
2-oxocaproate
pH 7.0, 30°C
Weissella confusa
0.17
-
2-oxooctanoate
pH 7.0, 30°C
Weissella confusa
0.45
-
2-oxobutyrate
pH 7.0, 30°C
Weissella confusa
0.6
-
2-Hydroxyisovalerate
pH 8.0, 30°C
Weissella confusa
0.62
-
2-hydroxyisocaproate
pH 8.0, 30°C
Weissella confusa
0.64
-
3-Phenyllactate
pH 8.0, 30°C
Weissella confusa
1.8
-
2-hydroxyvalerate
pH 8.0, 30°C
Weissella confusa
1.9
-
2-Hydroxyoctanoate
pH 8.0, 30°C
Weissella confusa
2.2
-
2-hydroxycaproate
pH 8.0, 30°C
Weissella confusa
4.5
-
pyruvate
pH 7.0, 30°C
Weissella confusa
100
-
L-lactate
pH 8.0, 30°C
Weissella confusa
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
additional information
at 1 mM no influence is observed with Mg2+, Ca2+, Co2+, Ni2+, and Cd2+ and very weak effects with Mn2+, Cu2+, and Zn2+
Weissella confusa
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
33000
-
4 * 33000, SDS-PAGE
Weissella confusa
125000
-
gel filtration
Weissella confusa
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
a 2-oxocarboxylate + NADH + H+
Weissella confusa
-
(2S)-2-hydroxycarboxylate + NAD+
-
-
r
Purification (Commentary) (protein specific)
Commentary
Organism
-
Weissella confusa
Source Tissue (protein specific)
Source Tissue
Commentary
Organism
Textmining
culture condition
maximum specific and volume activities are found at the end of the exponential growth phase, during further fermentation enzyme activity declines rapidly and is no longer detectable after 15 h
Weissella confusa
-
Storage Stability (protein specific)
Storage Stability
Organism
4°C, pH 6.5, concentrated enzyme solution, 10% loss of activity after 6 months
Weissella confusa
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
2-hydroxycaproate + NAD+
-
726750
Weissella confusa
2-oxocaproate + NADH + H+
-
-
-
r
2-hydroxyisocaproate + NAD+
the initial rate of the reduction of 2-oxoisocaproate is about seven times faster than the reverse reaction, the dehydrogenation of L-2-hydroxyisocaproate, measured in buffers at the respective pH optima
726750
Weissella confusa
2-oxoisocaproate + NADH + H+
-
-
-
r
2-hydroxyisovalerate + NAD+
-
726750
Weissella confusa
2-oxoisovalerate + NADH + H+
-
-
-
r
2-hydroxyoctanoate + NAD+
-
726750
Weissella confusa
2-oxooctanoate + NADH + H+
-
-
-
?
2-hydroxyvalerate + NAD+
-
726750
Weissella confusa
2-oxovalerate + NADH + H+
-
-
-
r
2-oxobutyrate + NADH + H+
-
726750
Weissella confusa
2-hydroxybutyrate + NAD+
-
-
-
r
2-oxocaproate + NADH + H+
-
726750
Weissella confusa
2-hydroxycaproate + NAD+
-
-
-
r
2-oxoisocaproate + NADH + H+
2-oxoisocaproate is the best substrate with the lowest KM-valus of 0.065 mM. Tthe initial rate of the reduction of 2-oxoisocaproate is about seven times faster than the reverse reaction, the dehydrogenation of L-2-hydroxyisocaproate, measured in buffers at the respective pH optima
726750
Weissella confusa
2-hydroxyisocaproate + NAD+
-
-
-
r
2-oxoisovalerate + NADH + H+
-
726750
Weissella confusa
2-hydroxyisovalerate + NAD+
-
-
-
r
2-oxooctanoate + NADH + H+
-
726750
Weissella confusa
2-hydroxyoctanoate + NAD+
-
-
-
r
2-oxovalerate + NADH + H+
-
726750
Weissella confusa
2-hydroxyvalerate + NAD+
-
-
-
r
3-phenyllactate + NAD+
-
726750
Weissella confusa
pyruvate + NADH + H+
-
-
-
r
a 2-oxocarboxylate + NADH + H+
-
726750
Weissella confusa
(2S)-2-hydroxycarboxylate + NAD+
-
-
-
r
a 2-oxocarboxylate + NADH + H+
various 2-oxocarboxylic acids are stereospecifically reduced to the corresponding (S)-2-hydroxycarboxylic acids. In the reverse reaction the NAD+-dependent dehydrogenation of L-2-hydroxycarboxylic acids is observed
726750
Weissella confusa
(2S)-2-hydroxycarboxylate + NAD+
-
-
-
r
L-lactate + NAD+
-
726750
Weissella confusa
pyruvate + NADH + H+
-
-
-
r
phenylpyruvate + NADH + H+
-
726750
Weissella confusa
3-phenyllactate + NAD+
-
-
-
r
pyruvate + NADH + H+
-
726750
Weissella confusa
L-lactate + NAD+
-
-
-
r
Subunits (protein specific)
Subunits
Commentary
Organism
tetramer
4 * 33000, SDS-PAGE
Weissella confusa
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
30
-
assay at
Weissella confusa
50
-
-
Weissella confusa
Temperature Range [°C] (protein specific)
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
Organism
30
50
the maximal temperature is around 50°C. At this temperature the reaction proceeded at about 3fold increase of the rate compared to 30°C
Weissella confusa
Temperature Stability [°C] (protein specific)
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
40
-
1 h, stable up to
Weissella confusa
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7
-
reduction of 2-oxoisocaproate
Weissella confusa
8
8.5
dehydrogenation of L-2-hydroxyisocaproate
Weissella confusa
pH Stability (protein specific)
pH Stability
pH Stability Maximum
Commentary
Organism
5.5
9
4°C, 24 h, stable
Weissella confusa
6.5
-
4°C, 6 months, concentrated enzyme solutions, 10% loss of activity
Weissella confusa
Other publictions for EC 1.1.1.337
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Synonyms
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
728749
Bao
An irreversible and kineticall ...
Weissella confusa
Protein J.
26
143-151
2007
-
-
-
-
-
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
2
-
-
2
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
1
-
-
-
-
-
-
662791
Rein
Dissimilation of cysteate via ...
Paracoccus pantotrophus, Paracoccus pantotrophus NKNCYSA / DSM 12449
Microbiology
151
737-747
2005
-
-
-
-
-
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
1
-
2
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
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-
-
-
-
-
-
-
-
1
-
2
-
-
-
-
-
-
-
-
-
-
1
1
-
-
-
728733
Chatterjee
High level expression and sing ...
Weissella confusa
Protein Expr. Purif.
39
137-143
2005
-
-
1
-
-
-
-
-
-
-
-
-
-
4
-
-
1
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
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-
-
1
-
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-
-
-
-
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-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
719359
Irimia
Methanoarchaeal sulfolactate d ...
Methanocaldococcus jannaschii
EMBO J.
23
1234-1244
2004
-
-
-
1
-
-
-
5
-
-
2
-
-
5
-
-
-
-
-
-
-
-
6
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
5
-
-
2
-
-
-
-
-
-
-
-
-
6
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
718960
Graupner
The first examples of (S)-2-hy ...
Methanothermus fervidus, Methanocaldococcus jannaschii
Biochim. Biophys. Acta
158
169-173
2001
-
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
5
-
3
-
-
-
-
-
-
-
1
-
-
-
-
-
-
1
-
-
-
-
-
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-
-
-
-
-
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-
-
-
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5
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
389520
Graupner
Identification of an archaeal ...
Methanothermus fervidus, Methanocaldococcus jannaschii
J. Bacteriol.
182
3688-3692
2000
-
-
2
-
-
-
-
10
-
-
-
-
-
8
-
-
-
-
-
-
-
-
12
-
2
-
-
-
-
-
-
-
4
-
-
-
-
-
2
4
-
-
-
-
-
-
10
-
-
-
-
-
-
-
-
-
-
-
-
12
-
-
-
-
-
-
-
-
-
-
2
2
-
-
-
728726
Feil
Modified substrate specificity ...
Weissella confusa
Protein Eng.
10
255-262
1997
-
-
-
-
9
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
7
-
2
-
-
-
54
-
-
-
-
-
-
-
-
-
-
-
-
9
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
7
-
-
-
-
54
-
-
-
-
-
-
-
-
54
54
728117
Niefind
Crystal structure of L-2-hydro ...
Weissella confusa
J. Mol. Biol.
251
256-281
1995
-
-
-
1
-
-
-
-
-
-
1
-
-
4
-
-
-
-
-
-
-
-
-
1
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
727405
Feil
Deletion variants of L-hydroxy ...
Weissella confusa
Eur. J. Biochem.
223
857-863
1994
-
-
-
-
9
-
-
35
-
-
-
-
-
4
-
-
1
-
-
-
-
-
4
-
2
1
-
-
35
1
-
-
-
-
-
-
-
-
-
-
-
9
-
-
-
-
35
-
-
-
-
-
-
-
1
-
-
-
-
4
-
1
-
-
35
1
-
-
-
-
-
-
-
35
35
727592
Lerch
Cloning, sequencing and expres ...
Weissella confusa
Gene
83
263-270
1989
-
1
1
-
-
-
-
-
-
-
-
-
-
11
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
1
1
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-
-
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-
-
-
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-
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-
-
-
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-
-
-
-
-
-
726750
Schütte
-
L-2-Hydroxyisocaproate dehydro ...
Weissella confusa
Appl. Microbiol. Biotechnol.
19
167-176
1984
-
-
-
-
-
-
7
15
-
1
2
1
-
1
-
-
1
-
-
1
-
1
17
1
1
2
1
1
-
2
-
2
2
-
-
-
-
-
-
2
-
-
-
-
7
-
15
-
1
2
1
-
-
-
1
-
1
-
1
17
1
2
1
1
-
2
-
2
-
-
-
-
-
-
-