Information on EC 5.4.99.14 - 4-Carboxymethyl-4-methylbutenolide mutase

Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Mark a special word or phrase in this record:
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


The expected taxonomic range for this enzyme is: Bacteria

EC NUMBER
COMMENTARY
5.4.99.14
-
RECOMMENDED NAME
GeneOntology No.
4-Carboxymethyl-4-methylbutenolide mutase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
4-Carboxymethyl-4-methylbut-2-en-1,4-olide = 4-carboxymethyl-3-methylbut-2-en-1,4-olide
show the reaction diagram
-
-
-
-
4-Carboxymethyl-4-methylbut-2-en-1,4-olide = 4-carboxymethyl-3-methylbut-2-en-1,4-olide
show the reaction diagram
isomerization does not involve actual methyl migration, but proceeds via the 1-methylbislactone
-
4-Carboxymethyl-4-methylbut-2-en-1,4-olide = 4-carboxymethyl-3-methylbut-2-en-1,4-olide
show the reaction diagram
mechanism
-
4-Carboxymethyl-4-methylbut-2-en-1,4-olide = 4-carboxymethyl-3-methylbut-2-en-1,4-olide
show the reaction diagram
isomerization does not involve actual methyl migration, but proceeds via the 1-methylbislactone
Rhodococcus rhodochrous N75
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
intramolecular transfer reaction
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
4-methylcatechol degradation (ortho cleavage)
-
Microbial metabolism in diverse environments
-
Toluene degradation
-
SYSTEMATIC NAME
IUBMB Comments
4-Carboxymethyl-4-methylbut-2-en-1,4-olide methylmutase
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4-Methyl-3-enelactone methyl isomerase
-
-
-
-
4-Methylmuconolactone methylisomerase
-
-
-
-
4-Methylmuconolactone methylisomerase
O51799
-
4-Methylmuconolactone methylisomerase
-
-
4-Methylmuconolactone methylisomerase
C5MR76
-
4-Methylmuconolactone methylisomerase
Pseudomonas reinekei MT1
-, C5MR76
-
-
Isomerase, 4-methyl-2-enelactone
-
-
-
-
methymuconolactone isomerase
-
-
methymuconolactone isomerase
Pseudomonas reinekei MT1
-
-
-
MLMI
O51799
-
MLMI
Pseudomonas reinekei MT1
C5MR76
-
-
MmlJ
Pseudomonas reinekei MT1
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
115300-03-3
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
basonym Alcaligenes eutrophus; JMP 134, cloned into Pseudomonas sp. B13 FR1
-
-
Manually annotated by BRENDA team
gene mmlI
UniProt
Manually annotated by BRENDA team
Gordonia rubripertincta N657
N657
-
-
Manually annotated by BRENDA team
strain MT1
-
-
Manually annotated by BRENDA team
strain MT1
UniProt
Manually annotated by BRENDA team
Pseudomonas reinekei MT1
strain MT1
-
-
Manually annotated by BRENDA team
Pseudomonas reinekei MT1
strain MT1
UniProt
Manually annotated by BRENDA team
Rhodococcus rhodochrous N5
N5
-
-
Manually annotated by BRENDA team
Rhodococcus rhodochrous N75
N75
-
-
Manually annotated by BRENDA team
Rhodococcus ruber N75
N75
-
-
Manually annotated by BRENDA team
strains BCN1, BCN2 and 4PH1
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
physiological function
O51799
the enzyme is involved in the modified ortho-cleavage pathway and methylmuconolactone degradation of Ralstonia eutropha strain JMP134
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(4R,5R)-5-chloro-3-methylmuconolactone
trans-3-methyldienelactone
show the reaction diagram
O51799
-
-
-
r
(4S)-4-methylmuconolactone
(4S)-3-methylmuconolactone
show the reaction diagram
Pseudomonas reinekei, Pseudomonas reinekei MT1
C5MR76
-
-
-
?
(4S,5S)-5-chloro-3-methylmuconolactone
trans-3-methyldienelactone
show the reaction diagram
O51799
-
-
-
r
1-methylbis-lactone
?
show the reaction diagram
Pseudomonas reinekei, Pseudomonas reinekei MT1
C5MR76
-
-
-
?
1-Methylbislactone
4-Carboxymethyl-3-methylbut-2-en-1,4-olide
show the reaction diagram
-
-
-
-
1-Methylbislactone
4-Carboxymethyl-3-methylbut-2-en-1,4-olide
show the reaction diagram
-
i.e. 1-methyl-3,7-dioxo-2,6-dioxabicyclo[3.3.0]octane
-
-
-
1-Methylbislactone
4-Carboxymethyl-3-methylbut-2-en-1,4-olide
show the reaction diagram
Rhodococcus rhodochrous N75
-
-
-
-
3-methylmuconolactone
4-methyl-3-oxoadipate enollactone
show the reaction diagram
Pseudomonas reinekei, Pseudomonas reinekei MT1
-
-
-
-
?
4-Carboxymethyl-4-methylbut-2-en-1,4-olide
4-Carboxymethyl-3-methylbut-2-en-1,4-olide
show the reaction diagram
-
-
-
-
4-Carboxymethyl-4-methylbut-2-en-1,4-olide
4-Carboxymethyl-3-methylbut-2-en-1,4-olide
show the reaction diagram
-
-
-
-
-
4-Carboxymethyl-4-methylbut-2-en-1,4-olide
4-Carboxymethyl-3-methylbut-2-en-1,4-olide
show the reaction diagram
-
-
-
-
4-Carboxymethyl-4-methylbut-2-en-1,4-olide
4-Carboxymethyl-3-methylbut-2-en-1,4-olide
show the reaction diagram
-
-
-
-
?
4-Carboxymethyl-4-methylbut-2-en-1,4-olide
4-Carboxymethyl-3-methylbut-2-en-1,4-olide
show the reaction diagram
C5MR76
-
-
-
?
4-Carboxymethyl-4-methylbut-2-en-1,4-olide
4-Carboxymethyl-3-methylbut-2-en-1,4-olide
show the reaction diagram
-
highly specific, accepts only (+)-(4S)-4-methylmuconolactone, i.e. 4-carboxymethyl-4-methylbut-2-en-1,4-olide and the putative reaction intermediate 1-methylbislactone
i.e. (-)-(4S)-3-methylmuconolactone
-
4-Carboxymethyl-4-methylbut-2-en-1,4-olide
4-Carboxymethyl-3-methylbut-2-en-1,4-olide
show the reaction diagram
Pseudomonas reinekei MT1
-
-
-
-
?
4-Carboxymethyl-4-methylbut-2-en-1,4-olide
4-Carboxymethyl-3-methylbut-2-en-1,4-olide
show the reaction diagram
Pseudomonas reinekei MT1
C5MR76
-
-
-
?
4-Carboxymethyl-4-methylbut-2-en-1,4-olide
4-Carboxymethyl-3-methylbut-2-en-1,4-olide
show the reaction diagram
Rhodococcus rhodochrous N5, Gordonia rubripertincta N657
-
-
-
-
4-Carboxymethyl-4-methylbut-2-en-1,4-olide
4-Carboxymethyl-3-methylbut-2-en-1,4-olide
show the reaction diagram
Rhodococcus rhodochrous N75
-
highly specific, accepts only (+)-(4S)-4-methylmuconolactone, i.e. 4-carboxymethyl-4-methylbut-2-en-1,4-olide and the putative reaction intermediate 1-methylbislactone
i.e. (-)-(4S)-3-methylmuconolactone
-
4-Carboxymethyl-4-methylbut-2-en-1,4-olide
4-Carboxymethyl-3-methylbut-2-en-1,4-olide
show the reaction diagram
Rhodococcus ruber N75
-
-
-
-
4-Carboxymethyl-4-methylbut-2-en-1,4-olide
?
show the reaction diagram
-
enzyme of the modified 3-oxoadipate pathway in the Gram-negative bacterium Alcaligenes eutrophus JMP 134
-
-
-
4-methyl-muconolactone
3-methyl-muconolactone
show the reaction diagram
O51799
-
-
-
r
additional information
?
-
-
not: isomeric methyl-substituted 4-carboxymethylbut-2-en-4-olides
-
-
-
additional information
?
-
Pseudomonas reinekei, Pseudomonas reinekei MT1
C5MR76
activity with 3-methyl-cis,cis-muconate is below the detection limit
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
4-Carboxymethyl-4-methylbut-2-en-1,4-olide
?
show the reaction diagram
-
enzyme of the modified 3-oxoadipate pathway in the Gram-negative bacterium Alcaligenes eutrophus JMP 134
-
-
-
4-methyl-muconolactone
3-methyl-muconolactone
show the reaction diagram
O51799
-
-
-
r
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
(+)-muconolactone
-
-
(-)1-Methyl-8-bromobislactone
-
weak
2,4-Dimethyl-2-enelactone
-
weak
-
2-Bromo-4-methylbutan-1,4-olide
-
weak
2-Bromobutan-1,4-olide
-
-
2-Methyl-2-enelactone
-
weak
-
2-Methylbutan-1,4-olide
-
-
3-Methyl-2-enelactone
-
weak
-
4-carboxymethylbut-2-en-4-olide
-
-
4-chloromercuribenzoate
C5MR76
remaining activity of 6% after incubation with 0.02 mM 4-chloromercuribenzoate for 30 min
4-chloromercuribenzoate
O51799
inhibition is reversible by DTT
4-methyl-2-enelactone
-
weak
-
4-Methylbut-3-en-1,4-olide
-
angelicalactone
4-methylbutan-1,4-olide
-
-
trans-dienelactone
-
weak
additional information
-
EDTA
-
additional information
-
not: isomeric methyl-substituted 4-carboxymethylbut-2-en-4-olides
-
additional information
O51799
no inhibition by EDTA
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.045
-
(4S)-4-methylmuconolactone
C5MR76
mutant enzyme Y39F, in phosphate buffer, 50 mM, pH 7.3
0.052
-
(4S)-4-methylmuconolactone
C5MR76
wild type enzyme, in phosphate buffer, 50 mM, pH 7.3
0.0972
-
(4S)-4-methylmuconolactone
C5MR76
mutant enzyme C67S, in phosphate buffer, 50 mM, pH 7.3
0.4583
-
(4S)-4-methylmuconolactone
C5MR76
mutant enzyme H52A, in phosphate buffer, 50 mM, pH 7.3
0.028
-
1-methyl-3,7-dioxo-2,6-dioxabicyclo[3.3.0]octane
-
-
0.17
-
1-methyl-bislactone
-
-
0.176
-
4-Carboxymethyl-4-methylbut-2-en-1,4-olide
-
-
0.23
-
4-Carboxymethyl-4-methylbut-2-en-1,4-olide
-
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
5.8
-
(4S)-4-methylmuconolactone
C5MR76
mutant enzyme Y39F, in phosphate buffer, 50 mM, pH 7.3
7.3
-
(4S)-4-methylmuconolactone
C5MR76
mutant enzyme C67S, in phosphate buffer, 50 mM, pH 7.3
24.2
-
(4S)-4-methylmuconolactone
C5MR76
wild type enzyme, in phosphate buffer, 50 mM, pH 7.3
47.2
-
(4S)-4-methylmuconolactone
C5MR76
mutant enzyme H52A, in phosphate buffer, 50 mM, pH 7.3
3.75
-
1-methyl-3,7-dioxo-2,6-dioxabicyclo[3.3.0]octane
-
-
98.5
-
4-Carboxymethyl-4-methylbut-2-en-1,4-olide
-
-
kcat/KM VALUE [1/mMs-1]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
100
-
(4S)-4-methylmuconolactone
C5MR76
mutant enzyme C67S, in phosphate buffer, 50 mM, pH 7.3; mutant enzyme H52A, in phosphate buffer, 50 mM, pH 7.3; mutant enzyme Y39F, in phosphate buffer, 50 mM, pH 7.3
293340
500
-
(4S)-4-methylmuconolactone
C5MR76
wild type enzyme, in phosphate buffer, 50 mM, pH 7.3
293340
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
28.8
-
C5MR76
toward 1-methylbis-lactone, in phosphate buffer, 50 mM, pH 7.3
57.3
-
C5MR76
toward (4S)-4-methylmuconolactone, in phosphate buffer, 50 mM, pH 7.3
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6.3
6.5
C5MR76
-
7.5
-
O51799
assay at
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6
8.5
-
active in this range, rapid decrease of activity above pH 8.5
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
25
-
-
assay at
25
-
O51799
assay at
30
-
-
assay at
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
Gordonia rubripertincta N657, Rhodococcus rhodochrous N5, Rhodococcus rhodochrous N75, Rhodococcus ruber N75
-
-
-
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
Rhodococcus rhodochrous N75
-
-
-
-
Manually annotated by BRENDA team
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
25000
-
O51799
gel filtration
75500
-
-
Rhodococcus rhodochrous, gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
homodimer
C5MR76
x-ray crystallography
homodimer
O51799
2 * 12900, about, sequence calculation
homodimer
Pseudomonas reinekei MT1
-
x-ray crystallography
-
monomer
-
1 * 40000, Alcaligenes eutrophus (JMP 134, cloned into Pseudomonas sp. B13 FR1), SDS-PAGE
tetramer
-
4 * 17000, Rhodococcus rhodochrous, SDS-PAGE
tetramer
Rhodococcus rhodochrous N75
-
4 * 17000, Rhodococcus rhodochrous, SDS-PAGE
-
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
in complex with (4S)-4-methylmuconolactone and (4S)-3-methylmuconolactone, sitting drop vapor diffusion method, at 19C, using 25% (w/v) polyethylene glycol 1500, 0.1 M MMT buffer, pH 5 (DL-malic acid, MES, and Tris base in a 1:2:2 molar ratio)
C5MR76
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
Chelating agents like EDTA or thiol-group modifying reagents like DTT stabilize
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
4C or -20C, highly purified enzyme is rather unstable
-
4C, 24 h, 10% loss of activity, highly purified enzyme, 5% loss of activity with addition of chelating agents like EDTA or thiol-group modifying reagents like DTT
-
-30C, slow loss of activity on prolonged storage
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
JMP 134, cloned into Pseudomonas sp. B13 FR1
-
MonoQ column chromatography
-
Strep-Tactin-Sepharose column chromatography
C5MR76
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
gene mmlI, DNA and amino acid sequence determination and analysis, expression in Escherichia coli strain BL21(DE3)
O51799
expressed in Escherichia coli JM109 cells
C5MR76
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
C67S
C5MR76
incubation of variant C67S with 4-chloromercuribenzoate does not significantly decrease its activity (above 65% of activity remaining after a 30-min incubation)
H26A
C5MR76
the mutant shows a marked decrease in catalytic activity (kcat) by two orders of magnitude
H52A
C5MR76
the mutation results in an increase in Km by a factor of 9, but only a small change in kcat
Y39F
C5MR76
the mutant displays reduced activity compared to the wild type enzyme
C67S
Pseudomonas reinekei MT1
-
incubation of variant C67S with 4-chloromercuribenzoate does not significantly decrease its activity (above 65% of activity remaining after a 30-min incubation)
-
H26A
Pseudomonas reinekei MT1
-
the mutant shows a marked decrease in catalytic activity (kcat) by two orders of magnitude
-
H52A
Pseudomonas reinekei MT1
-
the mutation results in an increase in Km by a factor of 9, but only a small change in kcat
-
Y39F
Pseudomonas reinekei MT1
-
the mutant displays reduced activity compared to the wild type enzyme
-