Information on EC 4.1.99.11 - benzylsuccinate synthase

for references in articles please use BRENDA:EC4.1.99.11
Word Map on EC 4.1.99.11
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results
Include results (AMENDA + additional results, but less precise)


The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea

EC NUMBER
COMMENTARY hide
4.1.99.11
-
RECOMMENDED NAME
GeneOntology No.
benzylsuccinate synthase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
benzylsuccinate = toluene + fumarate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
addition
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
toluene degradation to benzoyl-CoA (anaerobic)
-
-
toluene degradation
-
-
Toluene degradation
-
-
Metabolic pathways
-
-
Microbial metabolism in diverse environments
-
-
SYSTEMATIC NAME
IUBMB Comments
benzylsuccinate fumarate-lyase (toluene-forming)
A glycyl radical enzyme that is inhibited by benzyl alcohol, benzaldehyde, phenylhydrazine and is inactivated by oxygen.
CAS REGISTRY NUMBER
COMMENTARY hide
209264-18-6
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
related to Desulfosporosinus sp.
-
-
Manually annotated by BRENDA team
eubacteria, part of a methanogenic consortium
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
gene bssA
Q39VF1
UniProt
Manually annotated by BRENDA team
Geobacter metallireducens GS-15 / ATCC 53774 / DSM 7210
gene bssA
Q39VF1
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
archae, part of a methanogenic consortium
-
-
Manually annotated by BRENDA team
archae, part of a methanogenic consortium
-
-
Manually annotated by BRENDA team
no activity in Dechloromonas aromatica RCB
-
-
-
Manually annotated by BRENDA team
enzyme encoded as part of the tutE tutFDGH gene cluster
-
-
Manually annotated by BRENDA team
BssA; gene bssA
UniProt
Manually annotated by BRENDA team
BssA; gene bssA
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
metabolism
physiological function
-
a novel benzylsuccinate synthase alpha-subunit (bssA) sequence type in sedimentary microbial community originating from a tar-oil-contaminated aquifer at a former coal gasification plant
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2-fluorotoluene + fumarate
(2-fluorobenzyl)succinate
show the reaction diagram
-
-
-
?
3-fluorotoluene + fumarate
(3-fluorobenzyl)succinate
show the reaction diagram
-
-
-
?
4-fluorotoluene + fumarate
(4-fluorobenzyl)succinate
show the reaction diagram
-
-
-
?
benzaldehyde + fumarate
benzoylsuccinate
show the reaction diagram
-
-
-
?
benzylsuccinate
toluene + fumarate
show the reaction diagram
m-cresol + fumarate
(3-hydroxybenzyl)succinic acid
show the reaction diagram
m-xylene + fumarate
(3-methylbenzyl)succinic acid
show the reaction diagram
m-xylene-d6 + fumarate
(3-methylbenzyl)succinate-d6
show the reaction diagram
-
-
-
?
o-cresol + fumarate
(2-hydroxybenzyl)succinic acid
show the reaction diagram
o-xylene + fumarate
(2-methylbenzyl)succinic acid
show the reaction diagram
o-xylene-d10 + fumarate
(2-methylbenzyl)succinate-d10
show the reaction diagram
-
-
-
?
p-cresol + fumarate
(4-hydroxybenzyl)succinic acid
show the reaction diagram
p-xylene + fumarate
(4-methylbenzyl)succinic acid
show the reaction diagram
p-xylene-d10 + fumarate
(4-methylbenzyl)succinate-d10
show the reaction diagram
-
-
-
?
toluene + fumarate
benzylsuccinate
show the reaction diagram
toluene + maleate
benzylsuccinate
show the reaction diagram
-
-
-
?
[d3-methyl]toluene + fumarate
?
show the reaction diagram
-
deuterium is transferred from toluene to the C-3 pro-(S) position of benzylsuccinate, implying that the addition of toluene to the double bond of fumarate is syn
-
-
?
[d3-methyl]toluene + maleate
?
show the reaction diagram
-
addition of toluene occurs in anti fashion. Formation of the C-3 radical of the benzylsuccinate as an intermediate, in which rotation about the C-2-C-3 bond can occur to relieve the sterically unfavorable cis conformation
-
-
?
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
benzylsuccinate
toluene + fumarate
show the reaction diagram
toluene + fumarate
benzylsuccinate
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
flavin
-
indicated by fluorescence spectroscopy, chemical nature unknown
iron-sulfur centre
-
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
[4Fe-4S] center
-
[4 Fe-4S] clusters in benzylsuccinate synthase are identified via EPR and Mossbauer spectroscopy
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
-
tutE gene product (benzylsuccinate synthase activating enzyme) required for activity, changes in the tutE encoded protein (C72A and C79A) are able to prevent the action of benzylsuccinate synthase; tutH gene product required for activity, changes in the tutH encoded protein (G52A/K53R/S54A) are able to prevent the action of benzylsuccinate synthase, tutH gene product possibly involved in assembly of the enzyme subunits
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.11 - 0.35
Toluene
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0024
-
cell extract, 4°C
0.003
-
-
0.004
cell extract, pH 7.5, 30°C
0.0074
-
cell extract, 30°C
0.015
Q39VF1;
cell extract, pH 7.5, 30°C
0.017
cell extract, pH 7.5, 30°C
0.0195
cell extract, pH 7.5, 30°C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
additional information
PDB
SCOP
CATH
UNIPROT
ORGANISM
Thauera aromatica;
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6500
-
alpha, alpha, beta, gamma, 1 * 97000 + 1 * 97000 + 1 * 14500 + 1 * 6500, SDS-PAGE
7000
-
gamma subunit, determined by SDS-PAGE
10000
-
alpha, alpha, beta, gamma, 1 * 94000 + 1 * 90000 + 1 * 12000 + 1 * 10000, SDS-PAGE
12000
-
alpha, alpha, beta, gamma, 1 * 94000 + 1 * 90000 + 1 * 12000 + 1 * 10000, SDS-PAGE
14000
-
beta subunit, determined by SDS-PAGE
14500
-
alpha, alpha, beta, gamma, 1 * 97000 + 1 * 97000 + 1 * 14500 + 1 * 6500, SDS-PAGE
90000
-
alpha, alpha, beta, gamma, 1 * 94000 + 1 * 90000 + 1 * 12000 + 1 * 10000, SDS-PAGE
94000
-
alpha, alpha, beta, gamma, 1 * 94000 + 1 * 90000 + 1 * 12000 + 1 * 10000, SDS-PAGE
97000
-
alpha, alpha, beta, gamma, 1 * 97000 + 1 * 97000 + 1 * 14500 + 1 * 6500, SDS-PAGE
98000
-
98000, SDS-PAGE of the alpha subunit
100000
-
alpha subunit, determined by SDS-PAGE
220000
-
gel filtration
230000
-
determined by gel filtration, consistent with an alpha2beta2gamma2 subunit structure
260000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
-
98000, SDS-PAGE of the alpha subunit
heterotrimer
-
alpha, beta and gamma subunit
hexamer
tetramer
OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
extremely oxygen sensitive
rapidly inactivated by oxygen with a half-life of 20-30 s upon exposure to air
-
210663
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DEAE-Sepharose, hydroxyapatite, anion exchange chromatography, enzyme severely inactivated during purification
-
hydroxyapatite, gel filtration, 95% loss of specific activity after gel filtration
-
loss of activity during purification due to high oxygen sensitivity
-
using a Ni-NTA-agarose column
-
using a Strep-Tactin Superflow column
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli
-
expressed in Escherichia coli as a Strep-tagged fusion protein
-
expression in Escherichia coli BL21DE3 cells, the vectors pET-28b, pACYC-Duet, pET-Duet and pRSF-Duet are used
-
gene bssA, phylogenetic analysis
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C29S
-
tutG gene, strong loss of activity, engineered strain unable to grow with toluene as carbon source
C9Y/A10S
-
tutF gene, strong loss of activity, engineered strain unable to grow with toluene as carbon source
C29S
-
tutG gene, strong loss of activity, engineered strain unable to grow with toluene as carbon source
-
C9Y/A10S
-
tutF gene, strong loss of activity, engineered strain unable to grow with toluene as carbon source
-
E509A
site-directed mutagenesis, the mutation affects binding of fumarate
F384A
site-directed mutagenesis, the mutation affects binding of toluene
L390A
site-directed mutagenesis, the mutation affects binding of toluene
L491A
site-directed mutagenesis, the mutation affects binding of toluene
Q706A
site-directed mutagenesis, the mutation affects binding of fumarate
S827A
site-directed mutagenesis, the mutation affects binding of fumarate
V708A
site-directed mutagenesis, the mutation affects binding of toluene
Y829A
site-directed mutagenesis, the mutation affects binding of toluene
F384A
-
site-directed mutagenesis, the mutation affects binding of toluene
-
L390A
-
site-directed mutagenesis, the mutation affects binding of toluene
-
L491A
-
site-directed mutagenesis, the mutation affects binding of toluene
-
V708A
-
site-directed mutagenesis, the mutation affects binding of toluene
-
Y829A
-
site-directed mutagenesis, the mutation affects binding of toluene
-
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
the alpha subunit is purified and refolded from inclusion bodies
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
environmental protection
additional information
Show AA Sequence (189 entries)
Please use the Sequence Search for a specific query.