4.1.99.1: tryptophanase
This is an abbreviated version!
For detailed information about tryptophanase, go to the full flat file.
Word Map on EC 4.1.99.1
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4.1.99.1
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quinonoid
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transposase
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aldimine
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proteus
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phenol-lyase
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thermonuclease
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beta-elimination
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l-trp
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tryptophan-induced
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antitermination
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pyridoxal-p
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rho-dependent
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rapid-scanning
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phillips
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alvei
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analysis
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food industry
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biotechnology
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drug development
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medicine
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synthesis
- 4.1.99.1
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quinonoid
- transposase
-
aldimine
- proteus
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phenol-lyase
- thermonuclease
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beta-elimination
- l-trp
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tryptophan-induced
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antitermination
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pyridoxal-p
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rho-dependent
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rapid-scanning
-
phillips
- alvei
- analysis
- food industry
- biotechnology
- drug development
- medicine
- synthesis
Reaction
Synonyms
L-tryptophan indole-lyase, L-tryptophanase, TIL, tna2, TnaA, tnaA2, TNase, Tpase, Trpase, tryptophan indole lyase, tryptophan indole-lyase, tryptophan-indole lyase, tryptophanase, tryptophanase 2, VcTrpase
ECTree
4.1.99.1 tryptophanaseAdvanced search results
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results (Engineering
Engineering on EC 4.1.99.1 - tryptophanase
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PROTEIN VARIANTS 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
analysis
the metabolic enzyme tryptophanase (TPase) is used for biosensor construction, TPase is biotinylated so that it can be coupled with a molecular recognition element, such as an antibody, to develop an ELISA-like assay. This method is used for the detection of an antibody present in nM concentrations by the human nose. TPase can also be combined with the enzyme pyridoxal kinase (PKase) for use in a coupled assay to detect adenosine 5'-triphosphate (ATP). When ATP is present in the low mM concentration range, the coupled enzymatic system generates an odor that is easily detectable by the human nose. Biotinylated TPase can be combined with various biotinlabeled molecular recognition elements, thereby enabling a broad range of applications for this odor-based reporting system
C298S
the mutant displays reduced activity, subsequent to incubation at 2°C, the mutant Trpase loses about 90% of its activity
C352A/Q353A/Q354A
site-directed mutagenesis, the mutation delays TnaA focus disassembly in stationary phase
D363A/K366A
site-directed mutagenesis, the mutation does neither affect the enzyme localization nor the enzyme activity
D404A
site-directed mutagenesis, the mutation does neither affect the enzyme localization nor the enzyme activity
D42A/S43A/E44A/D45A
site-directed mutagenesis, the mutation delays TnaA focus disassembly in stationary phase
D49A/T52A/D53A/S54A
site-directed mutagenesis, the mutation delays TnaA focus disassembly in stationary phase
E17A/K20A/R21A
site-directed mutagenesis, the mutation does neither affect the enzyme localization nor the enzyme activity
E346A/E347A
site-directed mutagenesis, the mutation does neither affect the enzyme localization nor the enzyme activity
E384A/K387A/R392A
site-directed mutagenesis, the mutation delays TnaA focus disassembly in stationary phase
E416A
site-directed mutagenesis, the mutation alters TnaA focus formation during exponential growth
E416A/R419A
site-directed mutagenesis, the mutation alters TnaA focus formation during exponential growth
E437A/K440A/H441A
site-directed mutagenesis, the mutation does neither affect the enzyme localization nor the enzyme activity
E9A/R12A/R14A
F464A
site-directed mutagenesis, the mutation results in a 500fold decrease in kcat/Km for L-tryptophan, with less effect on the reaction of other nonphysiological elimination substrates. The mutation has no effect on the formation of quinonoid intermediates
H370A/D374A/Q375A
site-directed mutagenesis, the mutation does neither affect the enzyme localization nor the enzyme activity
H463F
K115A
site-directed mutagenesis, the mutation does neither affect the enzyme localization nor the enzyme activity
K156A
site-directed mutagenesis, the mutation does neither affect the enzyme localization nor the enzyme activity
K239A
site-directed mutagenesis, the mutation does neither affect the enzyme localization nor the enzyme activity
K270A
site-directed mutagenesis, the mutation delays TnaA focus disassembly in stationary phase
K33A/S34A
site-directed mutagenesis, the mutation does neither affect the enzyme localization nor the enzyme activity
K406A/K409A/Q410A
site-directed mutagenesis, the mutation does neither affect the enzyme localization nor the enzyme activity
K443A/E444A/N445A/N448A
site-directed mutagenesis, the mutation does neither affect the enzyme localization nor the enzyme activity
K450A
site-directed mutagenesis, the mutation does neither affect the enzyme localization nor the enzyme activity
K459A
site-directed mutagenesis, the mutation does neither affect the enzyme localization nor the enzyme activity
K467A/K469A/E470A
site-directed mutagenesis, the mutation does neither affect the enzyme localization nor the enzyme activity
K5A
site-directed mutagenesis, the mutation does neither affect the enzyme localization nor the enzyme activity
K5A/K115A/K156A/K239A/K450A/K459A
site-directed mutagenesis, the mutation does neither affect the enzyme localization nor the enzyme activity
N327A/D329A
site-directed mutagenesis, the mutation does neither affect the enzyme localization nor the enzyme activity
Q339A/Y340A/D343A
site-directed mutagenesis, the mutation does neither affect the enzyme localization nor the enzyme activity
Q429A/T430A/H431A/D433A
site-directed mutagenesis, the mutation alters TnaA focus formation during exponential growth
R27A/E28A/E29A
site-directed mutagenesis, the mutation delays TnaA focus disassembly in stationary phase
R403A
site-directed mutagenesis, the mutation does neither affect the enzyme localization nor the enzyme activity
R462A
site-directed mutagenesis, the mutation alters TnaA focus formation during exponential growth
R462A/H463A/T465A
site-directed mutagenesis, the mutation alters TnaA focus formation during exponential growth
S398A
S398A/R403A/D404A
T23A/R24A/Y26A
site-directed mutagenesis, the mutation does neither affect the enzyme localization nor the enzyme activity
T426A/Y427A/T428A
site-directed mutagenesis, the mutation alters TnaA focus formation during exponential growth
T453A/T455A/Y456A/E457A
site-directed mutagenesis, the mutation delays TnaA focus disassembly in stationary phase
T465A
site-directed mutagenesis, the mutation does neither affect the enzyme localization nor the enzyme activity
T60A/Q61A/S62A/Q64A
W330F
Y74F
the mutant displays reduced activity, the Y74F mutant has low activity at 25°C and its residual activity is further reduced by cooling
H463F
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site-directed mutagenesis, the mutant shows very low activity for elimination of indole but is still competent to form a quinonoid intermediate from L-tryptophan, it shows high activity with substrate beta-(benzimidazol-1-yl)-L-alanine
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C298S
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the mutant displays reduced activity, subsequent to incubation at 2°C, the mutant Trpase loses about 90% of its activity
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W330F
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the mutant displays reduced activity, subsequent to incubation at 2°C, the mutant Trpase loses about 90% of its activity
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Y74F
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the mutant displays reduced activity, the Y74F mutant has low activity at 25°C and its residual activity is further reduced by cooling
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F448H
site-directed mutagenesis, the imidazole of F448H TPL forms a hydrogen bond to the substrate, consistent with the histidine being capable of hydrogen bonding to the substrate in TIL
H458A
site-directed mutagenesis, the almost inactive mutant shows 99% reduced activity compared to the wild-type enzyme
Y72F
H463A
site-directed mutagenesis, the mutant shows 96% reduced activity compared to the wild-type
K270A
site-directed mutagenesis, the mutation possibly eliminates this cofactor-protein interaction in the enzyme, inactive mutant
R103A
site-directed mutagenesis, the mutation alters orientation of the cofactor pyridoxal 5'-phosphate, inactive mutant
K270A
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site-directed mutagenesis, the mutation possibly eliminates this cofactor-protein interaction in the enzyme, inactive mutant
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R103A
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site-directed mutagenesis, the mutation alters orientation of the cofactor pyridoxal 5'-phosphate, inactive mutant
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additional information
E9A/R12A/R14A
site-directed mutagenesis, the mutation alters TnaA focus formation during exponential growth
E9A/R12A/R14A
site-directed mutagenesis, the mutation delays TnaA focus disassembly in stationary phase
H463F
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the rate constant for quinonoid intermediate formation from L-Trp is about 10fold lower for H463F Trpase than for wild-type Trpase, but the rate constant for reaction of L-Met is similar for H463F Trpase and wild-type Trpase
H463F
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site-directed mutagenesis, the mutant shows very low activity for elimination of indole but is still competent to form a quinonoid intermediate from L-tryptophan, it shows high activity with substrate beta-(benzimidazol-1-yl)-L-alanine
H463F
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site-directed mutagenesis, the mutant shows very low activity for elimination of indole but is still competent to form a quinonoid intermediate from L-tryptophan, pH dependence of quinonoid intermediate formation, overview
H463F
site-directed mutagenesis, the mutation results in a 103fold decrease in tryptophan elimination activity and in a 1000fold decrease in tryptophan elimination activity and loss of the pKa of 6.0 in the pH dependence of kcat/Km, suggesting that His463 is that base. In contrast, kcat is pH-independent, demonstrating that only the correctly protonated form of the enzyme binds the substrate, and the enzyme-substrate complex does not undergo protonation
S398A
site-directed mutagenesis, the mutation alters TnaA focus formation during exponential growth
S398A
site-directed mutagenesis, the mutation delays TnaA focus disassembly in stationary phase
S398A/R403A/D404A
site-directed mutagenesis, the mutation alters TnaA focus formation during exponential growth
S398A/R403A/D404A
site-directed mutagenesis, the mutation delays TnaA focus disassembly in stationary phase
T60A/Q61A/S62A/Q64A
site-directed mutagenesis, the mutation alters TnaA focus formation during exponential growth
T60A/Q61A/S62A/Q64A
site-directed mutagenesis, the mutation delays TnaA focus disassembly in stationary phase
W330F
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as in wild type, upon cooling to 2°C, inactivation and dissociation of tetramer to dimer occurs, spectrofluorometry data
W330F
the mutant displays reduced activity, subsequent to incubation at 2°C, the mutant Trpase loses about 90% of its activity
Y72F
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mutation leads to a decrease in activity for L-tryptophan by 50000fold and to a considerable rearrangement of the active site. This rearrangement leads to an increase of room around the alpha -C atom of any bound amino acid, such that covalent binding of alpha -methyl-substituted amino acids becomes possible (which cannot be realized in wild-type Trpase)
Y72F
site-directed mutagenesis, the replacement leads to a drastic decrease in activity for L-tryptophan by 50000fold. On the other hand considerable activities are retained with respect to substrates bearing good leaving groups and to L-serine. Kinetics show a coexistence of induced fit and selected fit in the reaction mechanism of a mutant tryptophan indole lyase Y72F, analysis of interaction of the mutant tryptophan indole-lyase (TIL) from Proteus vulgaris Y72F with the transition state analogue, oxindolyl-L-alanine (OIA), with the natural substrate, L-tryptophan, and with a substrate S-ethyl-L-cysteine, overview. The change of Tyr 72 to Phe leads to a considerable increase in the enzyme affinity to OIA, and to a very strong decrease of kf and kr values
additional information
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transposon insertion in tnaA gene is associated with a decrease in both A549 cells adherence and biofilm formation by Escherichia coli
additional information
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construction of tnaA deletion and insertion mutant strains, overview
additional information
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synthesis of L-tryptophan from L-cysteine, pyridoxal 5'-phosphate, and indole by the recombinant enzyme expressed in Escherichia coli strain BL21(DE3), co-reaction with Pseudomonas sp. TS1138 cells that convert DL-2-amino-delta2-thiazoline-4-carboxylic acid to L-cysteine
additional information
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tnaA gene disruption of tryptophanase in Escherichia coli strain BL21(DE3) to prevent L-tryptophan and 5-hydroxy-L-tryptophan degradation for enhanced whole cell synthesis of 5-hydroxy-L-tryptophan using modified L-phenylalanine 4-hydroxylase, PAH-L101Y-W180F, from Chromobacterium violaceum in Escherichia coli, overview
additional information
construction of 42 TnaA variants: 6 truncated forms and 36 missense mutants in which different combinations of 83 surface-exposed residues are converted to alanine. A truncated TnaA protein containing only domains D1 and D3 (D1D3) localized to the pole. Mutations affecting the D1D3-to-D1D3 interface do not affect polar localization of D1D3 but do delay assembly of wild-type TnaA foci. In contrast, alterations to the D1D3-to-D2 domain interface produce diffuse localization of the D1D3 variant but do not affect the wild-type protein. Altering several surface-exposed residues decreases TnaA activity, implying that tetramer assembly may depend on interactions involving these sites. Changing any of three amino acids at the base of a loop near the catalytic pocket decreases TnaA activity and causes it to form elongated ovoid foci in vivo, indicating that the alterations affect focus formation and may regulate how frequently tryptophan reaches the active site. Mutant phenotypes, detailed overview
additional information
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construction of 42 TnaA variants: 6 truncated forms and 36 missense mutants in which different combinations of 83 surface-exposed residues are converted to alanine. A truncated TnaA protein containing only domains D1 and D3 (D1D3) localized to the pole. Mutations affecting the D1D3-to-D1D3 interface do not affect polar localization of D1D3 but do delay assembly of wild-type TnaA foci. In contrast, alterations to the D1D3-to-D2 domain interface produce diffuse localization of the D1D3 variant but do not affect the wild-type protein. Altering several surface-exposed residues decreases TnaA activity, implying that tetramer assembly may depend on interactions involving these sites. Changing any of three amino acids at the base of a loop near the catalytic pocket decreases TnaA activity and causes it to form elongated ovoid foci in vivo, indicating that the alterations affect focus formation and may regulate how frequently tryptophan reaches the active site. Mutant phenotypes, detailed overview
additional information
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construction of tnaA deletion and insertion mutant strains, overview
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additional information
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chimeric forms of Tna1/Tna2, thermal stability increases as the contnent of the N-terminal portion of Tna1 in the chimera increases
