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Enzyme Nomenclature
Information on EC 4.1.99.2 - tyrosine phenol-lyase
for references in articles please use BRENDA:EC4.1.99.2
Word Map on EC 4.1.99.2
- 4.1.99.2
- triptolide
- citrobacter
- freundii
-
preterm
-
two-photon
- herbicola
- wilfordii
-
quinonoid
-
topless
-
beta-elimination
-
tripterygium
-
nitroxide
-
aldimine
-
tempol
- tryptophanase
-
indole-lyase
-
nanorods
-
disaturated
-
phillips
- 3,4-dihydroxyphenyl-l-alanine
- synthesis
- degradation
- biotechnology
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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota
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EC NUMBER 
COMMENTARY 
4.1.99.2
-
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RECOMMENDED NAME
GeneOntology No.
tyrosine phenol-lyase
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
L-tyrosine + H2O = phenol + pyruvate + NH3
mechanism appears to require two bases, one of which abstracts the proton from the 2-position of the substrate to form a quinonoid intermediate and the second of which acting in concert with proton transfer from the first group to C-1 of the phenolic ring, abstracts the substrate hydroxyl group to facilitate cyclohexadienone formation and subsequent elimination of phenol
-
L-tyrosine + H2O = phenol + pyruvate + NH3
mechanism appears to require two bases, one of which abstracts the proton from the 2-position of the substrate to form a quinonoid intermediate and the second of which acting in concert with proton transfer from the first group to C-1 of the phenolic ring, abstracts the substrate hydroxyl group to facilitate cyclohexadienone formation and subsequent elimination of phenol
-
L-tyrosine + H2O = phenol + pyruvate + NH3
single-base racemization mechanism, proceeds with retention of configuration at C-beta
-
L-tyrosine + H2O = phenol + pyruvate + NH3
NH3 is the first substrate which interacts with bound pyridoxal 5'-phosphate, pyruvate is the second substrate
-
L-tyrosine + H2O = phenol + pyruvate + NH3
mechanism
-
L-tyrosine + H2O = phenol + pyruvate + NH3
mechanism
-
L-tyrosine + H2O = phenol + pyruvate + NH3
reversible cleavage of the Cbeta-Cgamma bond of L-tyrosine in the final stage of the beta-elimination, via intermediates: internal aldimine, external aldimine, quinoid, ketoquinoid, alpha-aminoacrylate, and internal aldimine, stepwise mechanism, overview
-
L-tyrosine + H2O = phenol + pyruvate + NH3
-
-
-
-
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
alpha,beta-elimination
racemization
transamination
alpha,beta-elimination
-
-
-
-
racemization
-
-
-
-
transamination
-
-
-
-
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
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SYSTEMATIC NAME
IUBMB Comments
L-tyrosine phenol-lyase (deaminating; pyruvate-forming)
A pyridoxal-phosphate protein. The enzyme cleaves a carbon-carbon bond, releasing phenol and an unstable enamine product that tautomerizes to an imine form, which undergoes a hydrolytic deamination to form pyruvate and ammonia. The latter reaction, which can occur spontaneously, can also be catalysed by EC 3.5.99.10, 2-iminobutanoate/2-iminopropanoate deaminase. The enzyme also slowly catalyses similar reactions with D-tyrosine, S-methyl-L-cysteine, L-cysteine, L-serine and D-serine.
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CAS REGISTRY NUMBER
COMMENTARY
9059-31-8
-
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ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Symbiobacterium sp.
-
33441, 33442, 33455, 33469, 650039, 650404, 651062, 652265, 653702, 653815, 664227, 664834, 665928, 678212, 678401, 679758, 713693, 713730, 726642, 726700, 727663
-
-
wild type enzyme and mutant enzymes R381A, R381V and R381I
UniProt
-
-
-
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GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
metabolism
-
tyrosine phenol lyase catalyses the biosynthesis of L-tyrosine/L-DOPA from ammonium, pyruvate and phenolic substrates through the reverse of alpha- and beta-elimination reaction
physiological function
-
hyper-3,4-dihydroxyphenyl-L-alanine-producing strain by a mutant TyrR, an activator of tpl, whereby highest productivity is obtained for the strain grown under non-induced conditions, which is 30fold higher than that obtained for tyrosine-induced wild-type cells
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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
1-amino-2-(4-hydroxyphenyl)-ethyl phosphinic acid + H2O
phenol + ?
-
-
-
?
2,5-difluorotyrosine + H2O
2,5-difluorophenol + pyruvate + NH3
-
-
-
?
2,6-difluorotyrosine + H2O
2,6-difluorophenol + pyruvate + NH3
-
-
-
?
2-fluorotyrosine + H2O
2-fluorophenol + pyruvate + NH3
-
-
-
?
3,4-dihydroxyphenyl-L-alanine + H2O
pyrocatechol + NH3 + pyruvate
-
-
-
-
?
3,5-difluorotyrosine + H2O
3,5-difluorophenol + pyruvate + NH3
-
-
-
?
3-chloro-L-tyrosine + H2O
?
-
substrate of mutant M379V, not of the wild-type enzyme
-
-
?
3-chlorotyrosine + H2O
3-chlorophenol + pyruvate + NH3
-
-
-
?
3-fluoro-L-tyrosine + H2O
3-fluoro-phenol + pyruvate + NH3
-
-
?
3-fluoro-L-tyrosine + H2O
3-fluorophenol + pyruvate + NH3
-
-
-
-
?
3-fluoro-L-tyrosine + H2O
?
-
substrate of mutant M379V, not of the wild-type enzyme
-
-
?
3-fluorotyrosine + H2O
3-fluorophenol + pyruvate + NH3
-
-
-
?
3-methoxy-L-tyrosine + H2O
?
-
substrate of mutant M379V, not of the wild-type enzyme
-
-
?
3-methyl-L-tyrosine + H2O
?
-
substrate of mutant M379V, not of the wild-type enzyme
-
-
?
L-Ala + pyridoxal phosphate
pyridoxamine phosphate + keto acid
-
-
-
-
ir
L-alanine + H2O
?
-
wild-type enzyme forms the Ala quinonoid intermediate when incubated with L-Ala
-
-
?
L-m-Tyr + pyridoxal phosphate
pyridoxamine phosphate + keto acid
-
-
-
-
ir
L-Phe + pyridoxal phosphate
pyridoxamine phosphate + keto acid
-
-
-
-
ir
L-Ser + pyridoxal phosphate
pyridoxamine phosphate + keto acid
-
-
-
-
ir
o-chlorophenol + H2O
?
-
substrate of mutant M379V, not of the wild-type enzyme
-
-
?
o-cresol + H2O
?
-
substrate of mutant M379V, not of the wild-type enzyme
-
-
?
o-methoxyphenol + H2O
?
-
substrate of mutant M379V, not of the wild-type enzyme
-
-
?
S-methyl-L-cysteine + H2O
?
Symbiobacterium sp.
-
385% of activity compared to L-tyrosine
-
?
3,4-dihydroxyphenyl-L-Ala + H2O
pyrocatechol + NH3 + pyruvate
-
-
-
?
3,4-dihydroxyphenyl-L-Ala + H2O
pyrocatechol + NH3 + pyruvate
-
i.e. L-dopa
-
?
3,4-dihydroxyphenyl-L-Ala + H2O
pyrocatechol + NH3 + pyruvate
-
i.e. L-dopa
-
?
beta-chloro-L-alanine + H2O
?
Symbiobacterium sp.
-
500% of activity compared to L-tyrosine
-
?
beta-chloro-L-alanine + H2O
?
-
500% of activity compared to L-tyrosine
-
?
catechol + pyruvate + NH3
3,4-dihydroxyphenyl-L-alanine
-
TPL is well-correlated to cytoplasmic 3,4-dihydroxyphenyl-L-alanine levels
-
-
r
catechol + pyruvate + NH3
3,4-dihydroxyphenyl-L-alanine
-
TPL is well-correlated to cytoplasmic 3,4-dihydroxyphenyl-L-alanine levels
-
-
r
D-tyrosine + H2O
phenol + pyruvate + NH3
Symbiobacterium sp.
-
at 7% the rate of L-tyrosine
-
?
D-tyrosine + H2O
phenol + pyruvate + NH3
-
at 7% the rate of L-tyrosine
-
?
L-2-aminoadipate + H2O
?
-
no activity with wild-type enzyme, activity with mutant enzyme R100T/V283R
-
-
?
L-2-aminoadipate + H2O
?
-
no activity with wild-type enzyme, activity with mutant enzyme R100T/V283R
-
-
?
L-3-4 dihydroxyphenylalanine + H2O
catechol + pyruvate + NH3
-
L-dopa
-
-
r
L-3-4 dihydroxyphenylalanine + H2O
catechol + pyruvate + NH3
-
L-DOPA, alpha-beta elimination
-
-
r
L-3-4 dihydroxyphenylalanine + H2O
catechol + pyruvate + NH3
-
L-dopa
-
-
r
L-Asp + H2O
formate + pyruvate + NH3
-
no activity with wild-type enzyme, activity with mutant enzyme R100T/V283R
-
?
L-Asp + H2O
formate + pyruvate + NH3
-
no activity with wild-type enzyme, activity with mutant enzyme R100T/V283R
-
?
L-Glu + H2O
?
-
no activity with wild-type enzyme, activity with mutant enzyme R100T/V283R
-
-
?
L-Glu + H2O
?
-
no activity with wild-type enzyme, activity with mutant enzyme R100T/V283R
-
-
?
L-Ser + phenol
L-Tyr + H2O
-
-
-
?
L-Ser + pyrocatechol
3,4-dihydroxyphenyl-L-Ala + H2O
-
-
i.e. L-dopa
?
L-Ser + pyrocatechol
3,4-dihydroxyphenyl-L-Ala + H2O
-
-
-
?
L-tyrosine + H2O
3,4-dihydroxyphenyl-L-alanine
-
TPL is well-correlated to cytoplasmic 3,4-dihydroxyphenyl-L-alanine levels
-
-
?
L-tyrosine + H2O
3,4-dihydroxyphenyl-L-alanine
-
TPL is well-correlated to cytoplasmic 3,4-dihydroxyphenyl-L-alanine levels
-
-
?
L-tyrosine + H2O
phenol + pyruvate + NH3
-
-
-
-
?
L-tyrosine + H2O
phenol + pyruvate + NH3
-
-
-
-
r
L-tyrosine + H2O
phenol + pyruvate + NH3
-
alpha-beta elimination
-
-
r
L-tyrosine + H2O
phenol + pyruvate + NH3
-
reversible cleavage of the Cbeta-Cgamma bond of L-tyrosine in the final stage of the beta-elimination, via intermediates: internal aldimine, external aldimine, quinoid, ketoquinoid, alpha-aminoacrylate, and internal aldimine, overview
-
-
r
L-tyrosine + H2O
phenol + pyruvate + NH3
-
alpha-beta elimination
-
-
r
pyrocatechol + pyruvate + NH3
3,4-dihydroxyphenyl-L-Ala + H2O
-
-
-
?
pyrocatechol + pyruvate + NH3
3,4-dihydroxyphenyl-L-Ala + H2O
-
-
-
?
pyrocatechol + pyruvate + NH3
3,4-dihydroxyphenyl-L-Ala + H2O
-
-
-
?
S-(o-nitrophenyl)-L-cysteine + H2O
?
Symbiobacterium sp.
-
250% of activity compared to L-tyrosine
-
?
S-(o-nitrophenyl)-L-cysteine + H2O
?
-
250% of activity compared to L-tyrosine
-
?
Tyr + H2O
phenol + pyruvate + NH3
-
L-Tyr
-
-
-
additional information
?
-
-
mechanism of alpha-proton isotope exchange in amino acids catalysed by tyrosine phenol-lyase
-
-
-
additional information
?
-
-
alanine, arginine, aspartate, cysteine, glutamine, glutamate, glycine, histidine, isoleucine, leucine, lysine, proline, serine, threonine, tryptophan are poor or no substrates, substrate specificity of purified tyrosine phenol lyase, overview
-
-
-
additional information
?
-
-
substrate 3-fluoro-L-tyrosine is converted only to the quinoid reaction intermediate, structure determination of 3-fluoro-L-tyrosine bound to enzyme mutant Y71F with open active site and mutant F448H with closed active site, overview
-
-
-
additional information
?
-
-
substrate specificity with 3-substituted tyrosines, the wild-type enzyme shows a narrow substrate spectrum, overview. Mutant M379V converts several L-tyrosine derivatives to the correspondent products
-
-
-
additional information
?
-
-
adaptive enzyme responsible for cells' growth on media with L-Tyr as the sole source of carbon
-
-
-
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NATURAL SUBSTRATES
NATURAL PRODUCTS
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
L-3-4 dihydroxyphenylalanine + H2O
catechol + pyruvate + NH3
-
L-dopa
-
-
r
L-3-4 dihydroxyphenylalanine + H2O
catechol + pyruvate + NH3
-
L-DOPA, alpha-beta elimination
-
-
r
L-3-4 dihydroxyphenylalanine + H2O
catechol + pyruvate + NH3
-
L-dopa
-
-
r
L-tyrosine + H2O
phenol + pyruvate + NH3
-
alpha-beta elimination
-
-
r
L-tyrosine + H2O
phenol + pyruvate + NH3
-
alpha-beta elimination
-
-
r
additional information
?
-
-
adaptive enzyme responsible for cells' growth on media with L-Tyr as the sole source of carbon
-
-
-
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
pyridoxal 5'-phosphate
-
2 pyridoxal 5'-phosphate binding sites per mol of enzyme; cofactor
pyridoxal 5'-phosphate
-
2 pyridoxal 5'-phosphate binding sites per mol of enzyme; cofactor
pyridoxal 5'-phosphate
-
contains 2 mol of pyridoxal phosphate per mol of enzyme; Km: 0.00152 mM
pyridoxal 5'-phosphate
-
the phosphate group changes from being monoanionic at low pH to dianionic at high pH
pyridoxal 5'-phosphate
-
dependent on, highest activity at 0.015-0.02 mM
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
K+
NH4+
additional information
-
divalent cations (Mg2+, Ca2+, Ba2+, and Sr2+) do not activate TPL
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INHIBITORS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
3,4-dihydroxyphenyl-L-alanine
-
inactivated by a Pictet-Spengler reaction between the cofactor and 3,4-dihydroxyphenyl-L-alanine, on treatment with excess pyridoxal-5'-phosphate the inactivated enzymes recovers over 80% of the original activity
phenol
-
In presence of 1 mM, only 23% of the original activity remained. The presence of phenol seriously affects its own production, through the inhibition of TPL
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
-
Vitreoscilla hemoglobin positive strains (vgb+ strains) have substantially higher TPL activity than the respective wild-type hosts or the strains bearing a comparable plasmid without vgb (the vgb- strains)
-
additional information
-
Vitreoscilla hemoglobin positive strains (vgb+ strains) have substantially higher TPL activity than the respective wild-type hosts or the strains bearing a comparable plasmid without vgb (the vgb- strains)
-
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
3.2
3,4-dihydroxyphenyl-L-alanine
-
wild type enzyme, in 0.1 M potassium phosphate buffer (pH 8.0) at 30°C
2
3-chloro-L-Ala
-
wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C
11.4
3-chloro-L-Ala
-
mutant enzyme D214N, in 50 mM potassium phosphate pH 8.0, at 25°C
13.3
3-chloro-L-Ala
-
mutant enzyme D214A, in 50 mM potassium phosphate pH 8.0, at 25°C
0.1
3-fluoro-L-Tyr
-
wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C
0.2
L-Tyr
-
wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C
0.22
L-tyrosine
-
mutant enzyme T15A, in 0.1 M potassium phosphate buffer (pH 8.0) at 30°C
0.24
L-tyrosine
-
wild type enzyme, in 0.1 M potassium phosphate buffer (pH 8.0) at 30°C
0.1
S-(o-nitrophenyl)-L-Cys
-
wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C
1.32
S-(o-nitrophenyl)-L-Cys
-
mutant enzyme D214A, in 50 mM potassium phosphate pH 8.0, at 25°C
1.86
S-(o-nitrophenyl)-L-Cys
-
mutant enzyme D214N, in 50 mM potassium phosphate pH 8.0, at 25°C
0.2
S-benzyl-L-Cys
-
wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C
5.1
S-benzyl-L-Cys
-
mutant enzyme D214N, in 50 mM potassium phosphate pH 8.0, at 25°C
7
S-benzyl-L-Cys
-
mutant enzyme D214A, in 50 mM potassium phosphate pH 8.0, at 25°C
3.4
S-methyl-L-Cys
-
wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C
13.4
S-methyl-L-Cys
-
mutant enzyme D214N, in 50 mM potassium phosphate pH 8.0, at 25°C
50
S-methyl-L-Cys
-
mutant enzyme D214A, in 50 mM potassium phosphate pH 8.0, at 25°C
additional information
additional information
-
pH-independent kinetic constants for substrates bearing small substituents
-
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.31
3,4-dihydroxyphenyl-L-alanine
-
wild type enzyme, in 0.1 M potassium phosphate buffer (pH 8.0) at 30°C
0.0065
3-chloro-L-Ala
-
mutant enzyme D214N, in 50 mM potassium phosphate pH 8.0, at 25°C
0.02
3-chloro-L-Ala
-
mutant enzyme D214A, in 50 mM potassium phosphate pH 8.0, at 25°C
0.7
3-chloro-L-Ala
-
wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C
1.4
3-fluoro-L-Tyr
-
wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C
3.5
L-Tyr
-
wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C
1.2
L-tyrosine
-
mutant enzyme T15A, in 0.1 M potassium phosphate buffer (pH 8.0) at 30°C
1.8
L-tyrosine
-
wild type enzyme, in 0.1 M potassium phosphate buffer (pH 8.0) at 30°C
0.024
S-(o-nitrophenyl)-L-Cys
-
mutant enzyme D214A, in 50 mM potassium phosphate pH 8.0, at 25°C
0.041
S-(o-nitrophenyl)-L-Cys
-
mutant enzyme D214N, in 50 mM potassium phosphate pH 8.0, at 25°C
5.1
S-(o-nitrophenyl)-L-Cys
-
wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C
7.4
S-(o-nitrophenyl)-L-cysteine
-
pH 8.0, 25°C, mutant enzyme E69D/K256R
0.0067
S-benzyl-L-Cys
-
mutant enzyme D214N, in 50 mM potassium phosphate pH 8.0, at 25°C
0.02
S-benzyl-L-Cys
-
mutant enzyme D214A, in 50 mM potassium phosphate pH 8.0, at 25°C
0.5
S-benzyl-L-Cys
-
wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C
0.0016
S-methyl-L-Cys
-
mutant enzyme D214N, in 50 mM potassium phosphate pH 8.0, at 25°C
0.0052
S-methyl-L-Cys
-
mutant enzyme D214A, in 50 mM potassium phosphate pH 8.0, at 25°C
0.9
S-methyl-L-Cys
-
wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C
additional information
additional information
-
pH-independent kinetic constants for substrates bearing small substituents
-
additional information
additional information
the turnover-numbers of the mutant enzyme R381I, R381V and R381A for the substrates S-(o-nitrophenyl)-L-Cys, 3-chloro-L-Ala, O-benzoyl-L-Ser and S-methyl-L-Cys are comparable to those of the wild-type enzyme
-
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Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
3
L-Asp
-
mutant enzyme D214A, in 50 mM potassium phosphate pH 8.0, at 25°C
3.5
L-Asp
-
wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C, using L-Tyr as a substrate; wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C, using S-(o-nitrophenyl)-L-Cys as a substrate
3.6
L-Asp
-
mutant enzyme D214N, in 50 mM potassium phosphate pH 8.0, at 25°C
5
L-Glu
-
wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C, using L-Tyr as a substrate; wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C, using S-(o-nitrophenyl)-L-Cys as a substrate
66
L-Glu
-
mutant enzyme D214N, in 50 mM potassium phosphate pH 8.0, at 25°C
100
L-Glu
-
mutant enzyme D214A, in 50 mM potassium phosphate pH 8.0, at 25°C
1.33
L-homoserine
-
wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C, using L-Tyr as a substrate; wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C, using S-(o-nitrophenyl)-L-Cys as a substrate
69
L-homoserine
-
mutant enzyme D214N, in 50 mM potassium phosphate pH 8.0, at 25°C
82
L-homoserine
-
mutant enzyme D214A, in 50 mM potassium phosphate pH 8.0, at 25°C
0.73
L-Met
-
wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C, using L-Tyr as a substrate; wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C, using S-(o-nitrophenyl)-L-Cys as a substrate
4.2
L-Met
-
mutant enzyme D214N, in 50 mM potassium phosphate pH 8.0, at 25°C
11
L-Met
-
mutant enzyme D214A, in 50 mM potassium phosphate pH 8.0, at 25°C
1.7
L-Phe
-
wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C, using L-Tyr as a substrate; wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C, using S-(o-nitrophenyl)-L-Cys as a substrate
17
L-Phe
-
mutant enzyme D214N, in 50 mM potassium phosphate pH 8.0, at 25°C
22
L-Phe
-
mutant enzyme D214A, in 50 mM potassium phosphate pH 8.0, at 25°C
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.0022
-
in strain S12TPL3-delta-hpd mutant (4-hydroxyphenylpyruvate dioxygenase knockout), harvested in logarithmic phase
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
8.2
8.5
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pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
6.5 - 9.2
Symbiobacterium sp.
-
pH 6.5: about 45% of maximal activity, pH 9.2: about 40% of maximal activity
7 - 9.5
Aeromonas phenologenes
-
pH 7.0: about 25% of maximal activity, pH 9.5: about 60% of maximal activity
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
30
45
80
45
-
mutant enzyme T15A, using 3,4-dihydroxyphenyl-L-alanine as a substrate
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TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
37 - 75
Symbiobacterium sp.
-
37°C: about 65% of maximal activity, 75°C: about 45% of maximal activity
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pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
-
wild-type S12 and its mutant strains S12TPL3 (optimized for phenol production)
additional information
additional information
-
optimization and evaluation of cultivation conditions, overview
additional information
-
optimization and evaluation of cultivation conditions, overview
-
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LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
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PDB
SCOP
CATH
UNIPROT
ORGANISM
Citrobacter freundii;
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
52000
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SUBUNITS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
homotetramer
tetramer
homotetramer
-
TPL is a homotetramer with four active sites, two per catalytic dimer, located at the monomer-monomer interface
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Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hanging drop vapour diffusion method with 50 mM potassium phosphate (pH 8.0), 2 mM DTT, 0.2 M KCl, and 32.5% (w/v) PEG2000 for the apo-enzyme, and 50 mM triethanolamine buffer (pH 8.0), containing 0.5 mM PLP, 2 mM DTT, 0.4-0.8 M KCl, and 35-38% (w/v) PEG5000 for the holo-enzyme
-
in complex with L-methionine and L-phenylalanine, wild type and mutant Y71F
-
purified wild-type enzyme in ternary complex with pyridine N-oxide and the L-Ala quinonoid reaction intermediate by soaking wild-type TPL crystals in the stabilization solution containing 40% w/v PEG 5000 MME, 50 mM triethanolamine, pH 8.0, 0.25 M KCl, 0.2 mM pyridoxal 5'-phosphate, 0.5 mM dithiothreitol, with 100 mM L-Ala and a saturating concentration of pyridine N-oxide for 20 s, and purified Y71F or F448H mutant enzymes with bound substrate in quinoid intermediate state by soaking the crystals in the same stabilizing solution but with addition of 10 mM 3-fluoro-L-Tyr for 30 s, X-ray diffraction structure determination and analysis at 2.05-2.25 A resolution
-
the quinonoid intermediates of tyrosine phenol-lyase with L-alanine and L-methionine are trapped in the crystalline state and their structures are determined at 1.9 A and 1.95 A resolution, respectively
-
tyrosine phenol-lyase complexed with 3-(4'-hydroxyphenyl)propionic acid
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pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
15 - 80
-
at 18°C the enzyme activity is about 20% of the maximal activity, when heated for 20 min, the wild type enzyme remains stable up to 55°C in a 0.1 M potassium phosphate buffer (pH 8.0), the half-inactivation temperature is calculated to be 62.2°C
40
50
70
80
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GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
liposomal encapsulation increases stability of the enzyme at 4°C for 3 weeks and at temperatures up to 61°C, little protection against trypsin and no protection against whole mouse plasma in vitro
-
mutant enzyme A13V loses 50% of its catalytic activity in 3 M urea, wild-type enzyme completely loses activity
-
the enzyme maintains its original activity at phenol concentrations below 75 mM
-
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
2°C, 0.02 M phosphate buffer + mercaptoethanol, pH 7.0, 1 week, 50% loss of activity
Aeromonas phenologenes
-
40°C, in the presence of 75 mM phenol at pH 8.5, 18h, no loss of activity
-
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
native enzyme 35fold to homogeneity by ammonium sulfate fractionation, dialysis, and hydrophobic interaction chromatography
-
partial
Resource Q ion exchange chromatography and Phenyl Superose Phenyl Superose column chromatography
-
wild-type protein and mutants T451A/A13V/E83K and T129I/A13V purified to homogeneity with purification yields of over 40%
-
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
expression in Escherichia coli, fusion to the lac and tac promoters, erpressed at high levels in the presence of isopropyl-beta-D-thiogalactopyranoside or lactose as inducer
-
gene tpl, expression of wild-type and mutant enzymes in Escherichia coli strain SVS 370
-
plasmid pHCE IIB-TPL ligated at 4°C for 24 h with a T4 DNA ligase, products electrotransformed into Escherichia coli JM83
-
Targeted gene disruptions are performed. Gene replacement vectors for the hpd, pobA, phhA, and aroP1 genes are created from pJQ200SK with different primers. Vectors are used to mutate the selected genes in Pseudomonas putida S12TPL3c by homologous recombination.
-
the plasmid pTZTPL is constructed for expression of the protein in Escherichia coli SVS370 cells
-
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
L-tyrosine addition only has a positive effect in terms of TPL induction for cell cultures harvested after 12 h
L-tyrosine induces the enzyme, liquid paraffin as oxygen vector has a stimulatory effect on the enzyme synthesis
TyrR is the transcriptional activator of Tpl. Amino acid substitutions of TyrR, V67A, Y72C and E201G in the N-terminal domain, N324D in the central domain, A503T in the C-terminal domain, all enhance Tpl expression
-
L-tyrosine addition only has a positive effect in terms of TPL induction for cell cultures harvested after 12 h
-
L-tyrosine addition only has a positive effect in terms of TPL induction for cell cultures harvested after 12 h
-
L-tyrosine addition only has a positive effect in terms of TPL induction for cell cultures harvested after 12 h
-
-
L-tyrosine addition only has a positive effect in terms of TPL induction for cell cultures harvested after 12 h
-
-
L-tyrosine induces the enzyme, liquid paraffin as oxygen vector has a stimulatory effect on the enzyme synthesis
-
L-tyrosine induces the enzyme, liquid paraffin as oxygen vector has a stimulatory effect on the enzyme synthesis
-
-
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
D214A
-
mutant does not catalyze the decomposition of L-Tyr and 3-fluoro-L-Tyr
D214N
-
mutant does not catalyze the decomposition of L-Tyr and 3-fluoro-L-Tyr
E69D/K256R
-
no activity with L-Tyr, turnover-number for S-(o-nitrophenyl)-L-cysteine is 1.5fold lower than the wild-type value, turnover-number for S-ethyl-L-Cys is 11fold lower than the wild-type value
F448H
H343A
-
all substrates for alpha,beta-elimination, except S-ethyl-L-Cys, exhibit lower turnover number values with the mutant enzyme than with the wild-type enzyme. The mutant shows slower rates of deuterium isotope exchange for L-Phe and L-Met than does the wild type enzyme. The turnover-number for 3-fluoro-L-Tyr is pH-dependent for the mutant enzyme, whereas it is pH-independent for the wild-type enzyme. His343 does play an important function in catalysis, possibly by facilitating the conformational change from an ‘open' to ‘closed' form when substrates bind
K256A
-
turnover-number for L-Tyr is 3500fold lower than wild-type value, turnover-number for S-(o-nitrophenyl)-L-cysteine is 560fold lower than the wild-type value, turnover-number for S-ethyl-L-Cys is 1560fold lower than the wild-type value, activity is not increased by addition of monovalent cations, K+, Na+, Li+, Rb+, or NH4+
K256H
-
turnover-number for L-Tyr is 26923fold lower than wild-type value, turnover-number for S-(o-nitrophenyl)-L-cysteine is 189fold lower than the wild-type value, turnover-number for S-ethyl-L-Cys is 443fold lower than the wild-type value, activity is not increased by addition of monovalent cations, K+, Na+, Li+, Rb+, or NH4+
K256R
-
turnover-number for L-Tyr is 29fold lower than wild-type value, turnover-number for S-(o-nitrophenyl)-L-cysteine is 30fold lower than the wild-type value, turnover-number for S-ethyl-L-Cys is 195fold lower than the wild-type value
M379V
-
changed substrate specificity; site-directed mutagenesis, the mutant is active with o-cresol, o-methoxyphenol, and o-chlorophenol, as well as 3-methyl-, 3-methoxy-, 3 -fluoro, and 3-chloro-L-tyrosine in contrast to the wild-type enzyme
N185A
-
2% residual activtiy with L-tyrosine or 3-fluoro-L-tyrosine, N185 stabilizes reaction intermediate
R381A
R381I
R381V
T124A
T124D
T124D/F448H
-
very little activity with L-tyrosine, significant activity with S-(o-nitrophenyl)-L-cysteine, S-alkyl-L-cysteine and beta-chloro-L-alanine
T15A
-
exhibits a 2fold improved activity towards 3,4-dihydroxyphenyl-L-alanine
Y71F
R100T/V283R
A13V
-
mutant exhibits higher temperature and denaturant stability than wild-type enzyme
T129I/A13V
-
increases the activity and thermal stability of the enzyme
T129I/A13V/E83K/T407A
-
increases the activity and thermal stability of the enzyme
T451A/A13V/E83K
-
increases the activity and thermal stability of the enzyme
T451A/A13V/E83K/T407A
-
increases the activity and thermal stability of the enzyme
additional information
F448H
-
very little activity with L-tyrosine, significant activity with S-(o-nitrophenyl)-L-cysteine, S-alkyl-L-cysteine and beta-chloro-L-alanine
F448H
-
very low activity with L-tyrosine, reduced activity with other substrates
F448H
-
site-directed mutagenesis, the mutant enzyme lacks the beta-elimination activity with L-Tyr or 3-fluoro-L-tyrosine as substrates, instead, it accumulates quinonoid intermediate
R381A
beta-elimination activity has been reduced by 0.0001fold compared to wild type enzyme
R381A
-
dramatic decrease in activity with L-tyrosine, but little effect on activity with other substrates
R381I
no detectable beta-elimination activity with L-Tyr as substrate
R381I
-
dramatic decrease in activity with L-tyrosine, but little effect on activity with other substrates
R381V
no detectable beta-elimination activity with L-Tyr as substrate
R381V
-
dramatic decrease in activity with L-tyrosine, but little effect on activity with other substrates
T124A
-
very little activity with L-tyrosine, significant activity with S-(o-nitrophenyl)-L-cysteine, S-alkyl-L-cysteine and beta-chloro-L-alanine
T124D
-
very little activity with L-tyrosine, significant activity with S-(o-nitrophenyl)-L-cysteine, S-alkyl-L-cysteine and beta-chloro-L-alanine
T124D
-
no detectable activity with L-tyrosine, but significant activity with other substrates with good leaving groups
Y71F
-
no activity for beta-elimination of L-Tyr. Can react with S-alkylcysteines, but these substrates exhibit a 1000-10000fold reduced turnover number compared to wild-type. For substrates with good leaving groups S-(o-nitrophenyl)-L-Cys, 3-chloro-L-Ala and O-benzoyl-L-Ser the mutant enzyme exhibits turnover numbers 1.85-7% those of the wild-type enzyme. Tyr 71 plays a dual role, both in cofactor binding in the absence of substrate and also as a general acid catalyst in the elimination of leaving groups from quinoid intermediates
Y71F
-
no detectable activity with L-tyrosine, but significant activity with other substrates with good leaving groups
Y71F
-
site-directed mutagenesis, the mutant enzyme lacks the beta-elimination activity with L-Tyr or 3-fluoro-L-tyrosine as substrates, instead, it accumulates quinonoid intermediate
R100T/V283R
-
increases the beta-elimination activity towards dicarboxylic amino acids, L-Asp, L-Glu and L-2-aminoadipate at least 10000-fold
R100T/V283R
-
increases the beta-elimination activity towards dicarboxylic amino acids, L-Asp, L-Glu and L-2-aminoadipate at least 10000-fold
-
additional information
-
substrate 3-fluoro-L-tyrosine is converted only to the quinoid reaction intermediate, structure determination of 3-fluoro-L-tyrosine bound to enzyme mutant Y71F with open active site and mutant F448H with closed active site, overview
additional information
-
sRNA-based knockdown of the two regulators and overexpression of the genes involved in the tyrosine biosynthetic pathway together with tyrosine phenol-lyase in Escherichia coli strains results in the production of phenol from glucose. The 18 engineered strains show significant differences in the production of tyrosine (i.e. the immediate precursor for phenol), TPL activity, and tolerance to phenol, overview. Engineered Escherichia coli strain BL21 produces phenol most efficiently by flask culture or fed-batch culture
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
biotechnology
degradation
synthesis
biotechnology
-
encapsulation of enzyme in wet nanoporous silica gels to selectively stabilize tertiary and quarternary protein conformations and to develop bioreactors and biosensors
biotechnology
-
tyrosine phenol lyase modifies and synthesizes natural and non-natural amino acids, synthesizes precursor of a large number of relevant compounds and plays an important role in phenolic waste treatment
biotechnology
-
tyrosine phenol lyase modifies and synthesizes natural and non-natural amino acids, synthesizes precursor of a large number of relevant compounds and plays an important role in phenolic waste treatment
-
degradation
Symbiobacterium sp.
-
removal and bioconversion of phenol in wastewater by a thermostable beta-tyrosinase
degradation
-
removal and bioconversion of phenol in wastewater by a thermostable beta-tyrosinase
-
synthesis
-
development of a multienzyme reactor with ec 4.1.99.2 and ec 4.1.1.25 for dopamine synthesis
synthesis
-
production of L-dopa, which is applied for the treatment of Parkinsonism
synthesis
-
major applications of tyrosine phenol lyase in the production of L-tyrosine and L-DOPA. Tyrosine phenol lyase also has the ability to modify and synthesize natural and non-natural amino acids
synthesis
-
tyrosine phenol lyase is an important enzyme for the synthesis of pharmaceutical aromatic amino acids, viz. L-tyrosine and L-DOPA
synthesis
-
major applications of tyrosine phenol lyase in the production of L-tyrosine and L-DOPA. Tyrosine phenol lyase also has the ability to modify and synthesize natural and non-natural amino acids
-
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LINKS TO OTHER DATABASES (specific for EC-Number 4.1.99.2)
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