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Information on EC 3.7.1.14 - 2-hydroxy-6-oxonona-2,4-dienedioate hydrolase and Organism(s) Escherichia coli and UniProt Accession P77044
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3.7.1.14 2-hydroxy-6-oxonona-2,4-dienedioate hydrolaseIUBMB Comments
This enzyme catalyses a step in a pathway of phenylpropanoid compounds degradation. The first step of the enzyme mechanism involves a reversible keto-enol tautomerization .
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The taxonomic range for the selected organisms is: Escherichia coli
The expected taxonomic range for this enzyme is: Bacteria, Archaea, Eukaryota
The expected taxonomic range for this enzyme is: Bacteria, Archaea, Eukaryota
Reaction Schemes
Synonyms
2-hydroxy-6-ketonona-2,4-diene-1,9-dioic acid 5,6-hydrolase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
2-hydroxy-6-ketonona-2,4-diene-1,9-dioic acid 5,6-hydrolase
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
(2Z,4E)-2-hydroxy-6-oxonona-2,4-diene-1,9-dioate + H2O = (2Z)-2-hydroxypenta-2,4-dienoate + succinate
(2Z,4E,7E)-2-hydroxy-6-oxonona-2,4,7-triene-1,9-dioate + H2O = (2Z)-2-hydroxypenta-2,4-dienoate + fumarate
a catalytic mechanism is proposed involving stabilisation of reactive intermediates and activation of a nucleophilic water molecule by Ser110
(2Z,4E)-2-hydroxy-6-oxonona-2,4-diene-1,9-dioate + H2O = (2Z)-2-hydroxypenta-2,4-dienoate + succinate
a catalytic mechanism is proposed involving stabilisation of reactive intermediates and activation of a nucleophilic water molecule by Ser110
(2Z,4E)-2-hydroxy-6-oxonona-2,4-diene-1,9-dioate + H2O = (2Z)-2-hydroxypenta-2,4-dienoate + succinate
although MCP hydrolases have a catalytic serine in the active site, the mechanism proceeds via a geminal diol, rather than an acyl-enzyme intermediate, reaction mechanism of the hydrolysis reaction, overview. MCP hydrolases accept alternative nucleophiles in addition to water, and accepts hydroxylamine in the C-C cleavage reaction. MhpC has a typical serine-hydrolase catalytic triad (Ser107, Asp228 and His256), but mechanistic studies indicate that the serine in the active site does not act as a nucleophile in the hydrolysis, but rather the reaction proceeds via general base catalysis.The serine in the active site might stabilise the oxyanion intermediate by hydrogen bonding
PATHWAY SOURCE
PATHWAYS
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-, -
SYSTEMATIC NAME
IUBMB Comments
(2E,4Z)-2-hydroxy-6-oxona-2,4-dienedioate succinylhydrolase
This enzyme catalyses a step in a pathway of phenylpropanoid compounds degradation. The first step of the enzyme mechanism involves a reversible keto-enol tautomerization [4].
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate + H2O
(2E)-2-hydroxypenta-2,4-dienoate + succinate
(2Z,4E)-2-hydroxy-6-oxonona-2,4-diene-1,9-dioate + H2O
(2Z)-2-hydroxypenta-2,4-dienoate + succinate
the enzyme also catalyses the reverse reaction of C-C hydrolysis, namely C-C bond formation
-
-
r
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate + H2O
(2E)-2-hydroxypenta-2,4-dienoate + succinate
2-hydroxy-6-keto-6-phenylhexa-2,4-dienoic acid + H2O
?
-
MhpC mutant is able to accept the 2-hydroxy-6-keto-6-phenylhexa-2,4-dienoic acid, on a shorter time scale
-
-
?
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate + H2O
(2E)-2-hydroxypenta-2,4-dienoate + succinate
-
-
-
?
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate + H2O
(2E)-2-hydroxypenta-2,4-dienoate + succinate
the enzyme is involved catabolism of 3-(3-hydroxyphenyl)propionate
-
-
?
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate + H2O
(2E)-2-hydroxypenta-2,4-dienoate + succinate
the enzyme is involved in 3-phenylpropanoate catabolism
-
-
?
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate + H2O
(2E)-2-hydroxypenta-2,4-dienoate + succinate
-
the enzyme is involved in 3-phenylpropanoate catabolism
-
-
?
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate + H2O
(2E)-2-hydroxypenta-2,4-dienoate + succinate
-
-
-
-
?
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate + H2O
(2E)-2-hydroxypenta-2,4-dienoate + succinate
-
the enzyme is involved in 3-phenylpropanoate catabolism
-
-
?
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate + H2O
(2E)-2-hydroxypenta-2,4-dienoate + succinate
the enzyme is involved in 3-phenylpropanoate catabolism
-
-
?
additional information
?
-
MCP hydrolases catalyse the C-C bond cleavage of compounds with a common structure, 2-hydroxy-6-oxohexa-2,4-dienoate with different substituents at the C-6 carbon
-
-
?
additional information
?
-
the enzyme is able to catalyse carbon-carbon bond formation. In addition to its natural substrate 2-hydroxy-6-oxonona-1,9-dienedioic acid, enzyme MhpC also hydrolyses various analogues and also the hydrolysis of ester bonds of monoethyl adipate and 4-nitrophenyl valerate. The H114A mutant of the enzyme also hydrolyses 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid, HOPDA, a substrate of enzyme BphD, EC 3.7.1.8. Incubation of monomethyl succinate and ethyl 2-hydroxypentadienoate with the wild-type freeze-dried MhpC in hexane result in C-C bond formation product
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate + H2O
(2E)-2-hydroxypenta-2,4-dienoate + succinate
(2Z,4E)-2-hydroxy-6-oxonona-2,4-diene-1,9-dioate + H2O
(2Z)-2-hydroxypenta-2,4-dienoate + succinate
the enzyme also catalyses the reverse reaction of C-C hydrolysis, namely C-C bond formation
-
-
r
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate + H2O
(2E)-2-hydroxypenta-2,4-dienoate + succinate
additional information
?
-
MCP hydrolases catalyse the C-C bond cleavage of compounds with a common structure, 2-hydroxy-6-oxohexa-2,4-dienoate with different substituents at the C-6 carbon
-
-
?
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate + H2O
(2E)-2-hydroxypenta-2,4-dienoate + succinate
the enzyme is involved catabolism of 3-(3-hydroxyphenyl)propionate
-
-
?
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate + H2O
(2E)-2-hydroxypenta-2,4-dienoate + succinate
the enzyme is involved in 3-phenylpropanoate catabolism
-
-
?
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate + H2O
(2E)-2-hydroxypenta-2,4-dienoate + succinate
-
the enzyme is involved in 3-phenylpropanoate catabolism
-
-
?
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate + H2O
(2E)-2-hydroxypenta-2,4-dienoate + succinate
-
the enzyme is involved in 3-phenylpropanoate catabolism
-
-
?
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate + H2O
(2E)-2-hydroxypenta-2,4-dienoate + succinate
the enzyme is involved in 3-phenylpropanoate catabolism
-
-
?
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0012
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme S40A
0.0025
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme H114A
0.004
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme N109A
0.004
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme N109H
0.0042
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme C261A
0.0055
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme H263A
0.0068
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, wild-type enzyme
0.0068
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°, wild-type enzyme
0.01
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme F173D
0.0175
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme S110G
0.0185
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme S110A
0.038
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme R188K
0.058
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme F173G
0.077
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme R188Q
0.125
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme W264G
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.001
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme S110G
0.0029
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme H263A
0.0054
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme S110A
0.016
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme D235A, approximate value from activity of the cell extract
0.08
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme N109H
0.1
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme R188Q
0.13
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme N109A
0.26
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme F173D
0.74
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme R188K
2 - 8
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, wild-type enzyme
2 - 8
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°, wild-type enzyme
2.7
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme W264G
2.9
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme S40A
4.5
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme H114A
7.5
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme F173G
12
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme C261A
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.057
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme S110G
0.29
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme S110A
1.4
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme R188Q
5.27
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme H263A
19
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme N109H
19
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme R188K
21
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme W264G
26
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme F173D
33
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme N109A
130
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme F173G
242
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme S40A
412
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, wild-type enzyme
1800
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme H114A
2900
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°C, mutant enzyme C261A
4118
(2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
pH 8.0, 25°, wild-type enzyme
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
physiological function
physiological function
the enzyme is involved in 3-phenylpropanoate catabolism
physiological function
the enzyme is involved catabolism of 3-(3-hydroxyphenyl)propionate
physiological function
the enzyme is involved in 3-phenylpropanoate catabolism
physiological function
-
the enzyme is involved in 3-phenylpropanoate catabolism
physiological function
the enzyme catalyse the hydrolysis of vinylogous 1,5-diketone meta-cleavage products generated during the biodegradation of various aromatic compounds
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
hanging drop vapor diffusion method, the X-ray structure of a succinate-H263A MhpC complex shows concerted movements in the positions of both Phe173 and Trp264 that line the approach to Arg188
hanging-drop vapour-diffusion method. The 2.1 A resolution X-ray structure of the native enzyme determined from orthorhombic crystals confirms that it is a member of the alpha/beta hydrolase fold family, comprising eight beta-strands interconnected by loops and helices. The 2.8 A resolution structure of the enzyme cocrystallized with the non-hydrolysable substrate analogue 2,6-diketo-nona-1,9-dioic acid confirms the location of the active site in a buried channel including Ser110, His263 and Asp235, postulated contributors to a serine protease-like catalytic triad in homologous enzymes
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
C261A
1.4fold decrease in kcat/Km for (2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
F173D
158fold decrease in kcat/Km for (2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
F173G
32fold decrease in kcat/Km for (2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
H114A
shows twofold-reduced catalytic efficiency, ruling out a catalytic role, but shows a fivefold-reduced KM for the natural substrate, and an ability to process an aryl-containing substrate, implying a role for His114 in positioning of the substrate
N109A
125fold decrease in kcat/Km for (2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
N109H
217fold decrease in kcat/Km for (2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
R188K
217fold decrease in kcat/Km for (2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
R188Q
2941fold decrease in kcat/Km for (2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
S110A
exhibits fast ketonisation, an intermediate phase, and slow C-C cleavage
S110G
exhibits fast ketonisation, an intermediate phase, and slow C-C cleavage
S40A
shows twofold reduced catalytic efficiency, but shows a very fast interconversion of dienol to dienolate forms of the substrate
W264G
196fold decrease in kcat/Km for (2E,4Z)-2-hydroxy-6-oxonona-2,4-dienedioate
H114A
-
MhpC mutant is able to accept the 2-hydroxy-6-keto-6-phenylhexa-2,4-dienoic acid, on a shorter time scale
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression of an engineered 3-(3-hydroxyphenyl)propionate catabolic cassette is analyzed in different gram-negative bacteria
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
3-(3-hydroxyphenyl)propionic acid, 3-(2-hydroxyphenyl)propionic acid, 3-(2,3-dihydroxyphenyl)propionic acid or hydrocinnamic acid or cis-3-(3-carboxyethyl)-3,5-cyclohexadiene-1,2-diol are poor inducers
-
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Burlingame, R.; Chapmen, P.J.
Catabolism of phenylpropionic acid and its 3-hydroxy-derivative by Escherichia coli
J. Bacteriol.
155
113-121
1983
Escherichia coli
Burlingame, R.P.; Wyman, L.; Chapman, P.J.
Isolation and characterization of Escherichia coli mutants defective for phenylpropionate degradation
J. Bacteriol.
168
55-64
1986
Escherichia coli
Diaz, E.; Ferrandez, A.; Garcia, J.L.
Characterization of the hca cluster encoding the dioxygenolytic pathway for initial catabolism of 3-phenylpropionic acid in Escherichia coli K-12
J. Bacteriol.
180
2915-2923
1998
Escherichia coli (P0ABW0)
Li, J.J.; Li, C.; Blindauer, C.A.; Bugg, T.D.
Evidence for a gem-diol reaction intermediate in bacterial C-C hydrolase enzymes BphD and MhpC from 13C NMR spectroscopy
Biochemistry
45
12461-12469
2006
Escherichia coli
Li, C.; Li, J.J.; Montgomery, M.G.; Wood, S.P.; Bugg, T.D.
Catalytic role for arginine 188 in the C-C hydrolase catalytic mechanism for Escherichia coli MhpC and Burkholderia xenovorans LB400 BphD
Biochemistry
45
12470-12479
2006
Escherichia coli (P77044)
Ferrandez, A.; Garcia, J.L.; Diaz, E.
Genetic characterization and expression in heterologous hosts of the 3-(3-hydroxyphenyl)propionate catabolic pathway of Escherichia coli K-12
J. Bacteriol.
179
2573-2581
1997
Escherichia coli (P77044)
Li, C.; Montgomery, M.G.; Mohammed, F.; Li, J.J.; Wood, S.P.; Bugg, T.D.
Catalytic mechanism of C-C hydrolase MhpC from Escherichia coli: kinetic analysis of His263 and Ser110 site-directed mutants
J. Mol. Biol.
346
241-251
2005
Escherichia coli (P77044)
Dunn, G.; Montgomery, M.G.; Mohammed, F., Coker, A.; Cooper, J.B.; Robertson, T.; Garcia, J.L.; Bugg, T.D.; Wood, S.P.
The structure of the C-C bond hydrolase MhpC provides insights into its catalytic mechanism
J. Mol. Biol.
346
253-265
2005
Escherichia coli (P77044), Escherichia coli
Siirola, E.; Frank, A.; Grogan, G.; Kroutil, W.
C-C hydrolases for biocatalysis
Adv. Synth. Catal.
355
1677-1691
2013
Escherichia coli (P77044)
-
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