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Information on EC 3.1.2.B5 - 3,4-hydroxyphenylacetyl-CoA thioesterase and Organism(s) Escherichia coli and UniProt Accession P76084

for references in articles please use BRENDA:EC3.1.2.B5
preliminary BRENDA-supplied EC number
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EC Tree
     3 Hydrolases
         3.1 Acting on ester bonds
             3.1.2 Thioester hydrolases
                3.1.2.B5 3,4-hydroxyphenylacetyl-CoA thioesterase
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This record set is specific for:
Escherichia coli
UNIPROT: P76084 not found.
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The taxonomic range for the selected organisms is: Escherichia coli
The enzyme appears in selected viruses and cellular organisms
Synonyms
PaaI, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SYSTEMATIC NAME
IUBMB Comments
3,4-dihydroxyphenylacetyl-CoA hydrolase
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
3,4-dihydroxyphenylacetyl-CoA + H2O
3,4-dihydroxyphenylacetate + CoA
show the reaction diagram
3,5-dihydroxyphenylacetyl-CoA + H2O
3,5-dihydroxyphenylacetate + CoA
show the reaction diagram
-
-
-
?
3-hydroxyphenylacetyl-CoA + H2O
3-hydroxyphenylacetate + CoA
show the reaction diagram
-
-
-
?
4-hydroxyphenylacetyl-CoA + H2O
4-hydroxyphenylacetate + CoA
show the reaction diagram
-
-
-
?
phenylacetyl-CoA + H2O
phenylacetate + CoA
show the reaction diagram
-
-
-
?
additional information
?
-
no activity with 4-hydroxybenzoyl-CoA, 4-hydroxybenzyl-CoA, 4-hydroxyphenacyl-CoA
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
3,4-dihydroxyphenylacetyl-CoA + H2O
3,4-dihydroxyphenylacetate + CoA
show the reaction diagram
the enzyme can liberate CoA from phenylacetyl-CoA and from hydroxyphenylacetyl-CoA substrates that are formed in the phenylacetate pathway
-
-
?
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.016 - 0.13
3,4-dihydroxyphenylacetyl-CoA
0.2
3,5-dihydroxyphenylacetyl-CoA
pH 8.1, 25°C, wild-type enzyme
0.021 - 0.23
3-hydroxyphenylacetyl-CoA
0.018 - 0.28
4-hydroxyphenylacetyl-CoA
0.0096
phenylacetyl-CoA
pH 8.1, 25°C, wild-type enzyme
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.076 - 100
3,4-dihydroxyphenylacetyl-CoA
9
3,5-dihydroxyphenylacetyl-CoA
pH 8.1, 25°C, wild-type enzyme
0.065 - 86
3-hydroxyphenylacetyl-CoA
0.17 - 79
4-hydroxyphenylacetyl-CoA
0.41
phenylacetyl-CoA
pH 8.1, 25°C, wild-type enzyme
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3 - 6250
3,4-dihydroxyphenylacetyl-CoA
45
3,5-dihydroxyphenylacetyl-CoA
pH 8.1, 25°C, wild-type enzyme
0.7 - 4095
3-hydroxyphenylacetyl-CoA
0.7 - 2257
4-hydroxyphenylacetyl-CoA
43
phenylacetyl-CoA
pH 8.1, 25°C, wild-type enzyme
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
SwissProt
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
58000
gel filtration
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homotetramer
4 *14720, ESI mass spectrometry
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
C-terminal His-tagged enzyme is prepared for crystallization. The N-terminal Met is replaced by Ser-Leu. Crystals of the selenomethionine enzyme are grown in 13% polyethylene glycol, 0.1 M [bis(2-hydroxyethyl)imino]-tris(hydroxymethyl)methane (pH 6.0) and 0.2 M lithium sulfate at 18°C using the hanging drop vapor diffusion method. They are incubated in the presence of 30% glycerol before being flash-frozen in liquid nitrogen. The crystals belong to trigonal space group P3(1)21 with a = b = 69.9 A and c = 117.2 A
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D16A
significantly reduced activity
D61A
small but detectable level of catalytic activity
E14A
reduced activity
H52A
1000fold decrease in the kcat/Km is observed with the 4-hydroxyphenylacetyl-CoA, and a 100fold decrease in the kcat/Km value is observed with the 3-hydroxyphenylacetyl-CoA or 3,4-dihydroxyphenylacetyl-CoA
N15A
reduced activity
N46A
significantly reduced activity
N46Q/D61E
no activity
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
0°C, pH 8.1, stored in 10 mM Tris buffer for several weeks without significant loss of activity
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Song, F.; Zhuang, Z.; Finci, L.; Dunaway-Mariano, D.; Kniewel, R.; Buglino, J.A.; Solorzano, V.; Wu, J.; Lima, C.D.
Structure, function, and mechanism of the phenylacetate pathway hot dog-fold thioesterase PaaI
J. Biol. Chem.
281
11028-11038
2006
Aromatoleum evansii (Q9F9V0), Aromatoleum evansii DSM 6898 (Q9F9V0), Escherichia coli (P76084)
Manually annotated by BRENDA team