Information on EC 4.2.1.34 - (S)-2-Methylmalate dehydratase

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The expected taxonomic range for this enzyme is: Bacteria

EC NUMBER
COMMENTARY hide
4.2.1.34
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RECOMMENDED NAME
GeneOntology No.
(S)-2-Methylmalate dehydratase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
(S)-2-Methylmalate = 2-methylfumarate + H2O
show the reaction diagram
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
condensation
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elimination
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
C5-Branched dibasic acid metabolism
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glutamate and glutamine metabolism
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L-glutamate degradation VI (to pyruvate)
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Metabolic pathways
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SYSTEMATIC NAME
IUBMB Comments
(S)-2-methylmalate hydro-lyase (2-methylfumarate-forming)
Also hydrates fumarate to (S)-malate.
CAS REGISTRY NUMBER
COMMENTARY hide
9027-94-5
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Aerobic bacterium
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Manually annotated by BRENDA team
YG1002
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Manually annotated by BRENDA team
YG1002
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Manually annotated by BRENDA team
YG-0504
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Manually annotated by BRENDA team
YG-0504
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
YG-0601
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Manually annotated by BRENDA team
YG-0601
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Manually annotated by BRENDA team
IAM 1133
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Manually annotated by BRENDA team
IAM 1133
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Manually annotated by BRENDA team
strain 1R
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Manually annotated by BRENDA team
strain 1R
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Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(L)-citramalate
mesaconate + H2O
show the reaction diagram
(S)-malate
fumarate + H2O
show the reaction diagram
Citramalate + H2O
Mesaconate
show the reaction diagram
Fumarate + H2O
L-Malate
show the reaction diagram
mesaconate
citramalate + H2O
show the reaction diagram
additional information
?
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
additional information
?
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
D-malate
PCMB
Aerobic bacterium
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0.15 mM, complete inhibition. 80% protection by 5 mM S-citramalate
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2,3-dimercapto-1-propanol
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about half as effective as L-Cys
2-mercaptoethylamine
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at pH 8.2-8.4 as effective as L-Cys
acetate
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activity is higher in extracts of cells grown on acetate
D-Cys
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at pH 8.2-8.4 as effective as L-Cys
DL-homocysteine
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at pH 8.2-8.4 as effective as L-Cys
L-Cys
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required. The optimal concentration of L-Cys is 0.5-2.0 mM
sulfhydryl compounds
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required
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2 - 4
(+)-citramalate
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0.11 - 0.95
fumarate
2.4
L-citramalate
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6.7
L-Malate
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0.07 - 0.59
mesaconate
0.45
S-citramalate
Aerobic bacterium
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0.56
S-malate
Aerobic bacterium
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.1
Aerobic bacterium
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additional information
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of component I and II
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5 - 7
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in presence of Cys
8 - 8.5
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in presence of 2-mercaptoethanol
8.5
Aerobic bacterium
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with mesaconate as substrate
8.7
Aerobic bacterium
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with fumarate as substrate
9
Aerobic bacterium
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S-malate as substrate
9.1
Aerobic bacterium
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with S-citramalate as substrate
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.7 - 7.7
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about 50% of maximal activity at pH 7.1 and at pH 9.3, in presence of Cys
7.1 - 9.3
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about 50% of maximal activity at pH 7.1 and at pH 9.3, in presence of 2-mercaptoethanol
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
160000
Aerobic bacterium
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gel filtration
additional information
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the enzyme consists of two components: component I has MW 45000 and component II has MW 310000, determined by gel filtration. A combination of both components gives the active enzyme. At pH 7.5 or below component II partially dissociates into species with molecular weights of about 165000, 82000, and 37000. At pH 9.0 these species reassociate to form active component II of about the original MW. The observed species may represent a monomer, dimer, tetramer and octamer of a single subunit
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7 - 11
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component I is stable
5668
7 - 8
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4C, 48 h, about 20% loss of activity of component II
5668
7.5
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at pH 7.5 or below component II partially dissociates into species with molecular weights of about 165000 Da, 82000 Da, and 37000 Da. At pH 9.0 these species reassociate to form active component II of about the original MW. The observed species may represent a monomer, dimer, tetramer and octamer of a single subunit
5666
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
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2 d, component II is stable in presence of mercaptoethanol, Fe2+, and component I
55
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5 min, component II is stable in presence of mercaptoethanol and Fe2+
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
freezing and thawing does not significantly decrease the activity of component I, 10-15% loss of activity of component II
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OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
4C, 90% loss of activity after 24 h in air. Reactivation by Fe2+ and 2-mercaptoethanol
Aerobic bacterium
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5667
inactivation by exposure to oxygen, full activation by anaerobic incubation with a sulfhydryl compound for 1-2 hours at 37C
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5665
inactivation by oxygen. Reactivation by incubating the two protein components with a reducing agent and Fe2+ in absence of oxygen. Mn2+ inhibits activation. Effect of temperature and pH on activation
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5669
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-10C, pH 8.4, protein concentration above 1 mg/ml in 0.1 M Tris-HCl buffer, component I is stable for months
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-196C, activity of component II remains constant for at least 8 months
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-20C, in presence of 20% (NH4)2SO4 or 10% glycerol in air, about 80-90% can be reactivated even after several months
Aerobic bacterium
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4C, 70% loss of activity after 1 week under N2
Aerobic bacterium
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
component I and II
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component I and II; whole enzyme
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