4.1.99.2: tyrosine phenol-lyase
This is an abbreviated version!
For detailed information about tyrosine phenol-lyase, go to the full flat file.
Word Map on EC 4.1.99.2
-
4.1.99.2
-
triptolide
-
citrobacter
-
threatened
-
preterm
-
two-photon
-
luminescence
-
freundii
-
topless
-
tripterygium
-
wilfordii
-
nitroxide
-
quinonoid
-
hook
-
labour
-
erwinia
-
herbicola
-
co-repressors
-
nanorods
-
tempol
-
beta-elimination
-
disaturated
-
3,4-dihydroxyphenyl-l-alanine
-
tryptophanase
-
aldimine
-
indole-lyase
-
phillips
-
tocolysis
-
triepoxide
-
synthesis
-
photoluminescence
-
degradation
-
biotechnology
- 4.1.99.2
- triptolide
- citrobacter
-
threatened
-
preterm
-
two-photon
-
luminescence
- freundii
-
topless
- tripterygium
- wilfordii
-
nitroxide
-
quinonoid
-
hook
-
labour
- erwinia
- herbicola
-
co-repressors
-
nanorods
-
tempol
-
beta-elimination
-
disaturated
- 3,4-dihydroxyphenyl-l-alanine
- tryptophanase
-
aldimine
-
indole-lyase
-
phillips
-
tocolysis
-
triepoxide
- synthesis
-
photoluminescence
- degradation
- biotechnology
Reaction
Synonyms
beta-tyrosinase, Fn-TPL, L-tyrosine phenol-lyase, phenol-lyase, tyrosine, TnaA, TPL, tyrosine phenol lyase, tyrosine phenol-lyase, tyrosine-phenol lyase
ECTree
4.1.99.2 tyrosine phenol-lyaseAdvanced search results
results (
results (Engineering
Engineering on EC 4.1.99.2 - tyrosine phenol-lyase
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
top
PROTEIN VARIANTS 
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
F448A
site-directed mutagenesis, the mutant shows a 104fold reduced activity with L-tyrosine compared to wild-type, stopped-flow kinetics of enzyme mutant F448A. The mutant F448A TPL forms quinonoid intermediates from L-tyrosine and S-ethyl-L-cysteine with rate constants similar to those of wild-type TPL, and can form an aminoacrylate intermediate from S-ethyl-L-cysteine but not L-tyrosine, with a rate constant similar to that of wild-type TPL
F448H
F448L
site-directed mutagenesis, the mutant shows a 103fold reduced activity with L-tyrosine compared to wild-type
F449A
site-directed mutagenesis, the mutant shows a 104fold reduced activity with L-tyrosine compared to wild-type
H343A
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
N185A
-
2% residual activtiy with L-tyrosine or 3-fluoro-L-tyrosine, N185 stabilizes reaction intermediate
R381A
R381I
R381V
S51A
site-directed mutagenesis, the mutation leads to a decrease of the kcat/Km parameter for reactions with L-tyrosine and 3-fluoro-L-tyrosine by three orders of magnitude, compared to the wild-type enzyme, phenotype, overview. Influence of replacement of Ser51 by Ala on the kinetic parameters of TPL reactions with inhibitory L-phenylalanine and L-methionine, kinetics and structures, overview
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
F448H
site-directed mutagenesis, enzyme mutant crystal structure with bound 3-fluoro-L-tyrosine, tense and closed conformation of F448H TPL quinonoid complex with the ligand, overview
F448H
site-directed mutagenesis, enzyme mutant crystal structure with bound 3-fluoro-L-tyrosine, tense and closed conformation of F448H TPL quinonoid complex with the ligand, overview. Mutant F448H TPL has very low catalytic activity with L-tyrosine compared to wild-type
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
H343A
site-directed mutagenesis, the mutant shows altered kinetics compared to wild-type enzyme
M379V
-
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
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
R381A
site-directed mutagenesis, the mutant shows very low remaining activity compared to wild-type
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
T124A
site-directed mutagenesis, the mutant shows very low remaining activity compared to wild-type
T124D
-
no detectable activity with L-tyrosine, but significant activity with other substrates with good leaving groups
T124D
very little activity with L-tyrosine, significant activity with S-(o-nitrophenyl)-L-cysteine, S-alkyl-L-cysteine and beta-chloro-L-alanine
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
Y71F
site-directed mutagenesis, enzyme mutant crystal structure with bound 3-fluoro-L-tyrosine, open conformation of Y71F TPL quinonoid complex with the ligand, overview
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
L-dihydroxyphenylalanine (DOPA) is biosynthesized by a tyrosine-phenol lyase from catechol, pyruvate, and ammonia in recombinant Escherichia coli expressing the enzyme, and the biosynthesized amino acid is directly incorporated into proteins. Three biochemical experiments with mutant proteins containing DOPA confirm the genetic incorporation of biosynthesized DOPA, and reveal its potential for various biochemical applications
additional information
-
L-dihydroxyphenylalanine (DOPA) is biosynthesized by a tyrosine-phenol lyase from catechol, pyruvate, and ammonia in recombinant Escherichia coli expressing the enzyme, and the biosynthesized amino acid is directly incorporated into proteins. Three biochemical experiments with mutant proteins containing DOPA confirm the genetic incorporation of biosynthesized DOPA, and reveal its potential for various biochemical applications
-
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
additional information
the tyrosine phenol lyase (TPL) catalyzed synthesis of L-DOPA is regarded as one of the most economic routes for L-DOPA synthesis, usage in a fed-batch approach with sodium pyruvate, dicatechol and ammonium acetate. In addition, L-DOPA crystals are exogenously added to inhibit cell encapsulation by the precipitated product. The final L-DOPA concentration reaches higher than 120 g/l with dicatechol conversion more than 96% in a 15-l stirred tank, demonstrating the great potential of Fn-TPL for industrial production of L-DOPA. Optimization of reaction conditions and upscaling of the recombinant enzyme TPL from Fusobacterium nucleatum in enzyme-expressing Escherichia coli cells, detailed overview
additional information
Fusobacterium nucleatum subsp. nucleatum ATCC 25586 / CIP 101130 / JCM 8532 / LMG 13131
-
the tyrosine phenol lyase (TPL) catalyzed synthesis of L-DOPA is regarded as one of the most economic routes for L-DOPA synthesis, usage in a fed-batch approach with sodium pyruvate, dicatechol and ammonium acetate. In addition, L-DOPA crystals are exogenously added to inhibit cell encapsulation by the precipitated product. The final L-DOPA concentration reaches higher than 120 g/l with dicatechol conversion more than 96% in a 15-l stirred tank, demonstrating the great potential of Fn-TPL for industrial production of L-DOPA. Optimization of reaction conditions and upscaling of the recombinant enzyme TPL from Fusobacterium nucleatum in enzyme-expressing Escherichia coli cells, detailed overview
-
