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Enzyme Nomenclature
Information on EC 4.1.99.1 - tryptophanase
for references in articles please use BRENDA:EC4.1.99.1
Word Map on EC 4.1.99.1
- 4.1.99.1
-
catabolite
-
quinonoid
- transposase
-
aldimine
- proteus
-
phenol-lyase
-
beta-elimination
- l-trp
- thermonuclease
-
tryptophan-induced
-
antitermination
-
pyridoxal-p
- d-serine
-
phillips
-
plp-dependent
- alvei
-
rapid-scanning
-
rho-dependent
- drug development
- medicine
- biotechnology
- food industry
- synthesis
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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota
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EC NUMBER 
COMMENTARY 
4.1.99.1
-
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RECOMMENDED NAME
GeneOntology No.
tryptophanase
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
L-tryptophan + H2O = indole + pyruvate + NH3
-
-
-
-
L-tryptophan + H2O = indole + pyruvate + NH3
mechanism that requires two catalytic bases
-
L-tryptophan + H2O = indole + pyruvate + NH3
in the reverse direction NH4+ interacts with bound pyridoxal 5'-phosphate to form an imine. Pyruvate is the second substrate, indole the third. alpha-Aminoacrylate functions as a common enzyme-bound intermediate in both synthetic and degradative reactions
-
L-tryptophan + H2O = indole + pyruvate + NH3
quinonoid intermediate formation involving Tyr71 and Cys298, mechanism, active site preorganization, overview
-
L-tryptophan + H2O = indole + pyruvate + NH3
reaction mechanism, overview
-
-
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
beta-elimination
elimination
-
-
alpha,beta-position of amino acid; beta-position of amino acid
-
replacement
-
-
beta-position of amino acid
-
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
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SYSTEMATIC NAME
IUBMB Comments
L-tryptophan indole-lyase (deaminating; pyruvate-forming)
A pyridoxal-phosphate protein, requiring K+. The enzyme cleaves a carbon-carbon bond, releasing indole 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. Also catalyses 2,3-elimination and beta-replacement reactions of some indole-substituted tryptophan analogues of L-cysteine, L-serine and other 3-substituted amino acids.
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CAS REGISTRY NUMBER
COMMENTARY
9024-00-4
-
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ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
37334, 37335, 37336, 37337, 37339, 37340, 37341, 37342, 37348, 37351, 37354, 37355, 37357, 37358, 37359, 37360, 37362, 37364, 650374, 650435, 650757, 651028, 663488, 664076, 677332, 681216, 681998, 691648, 698614, 703486, 704093, 704722, 704847, 705578, 705580, 713955, 715733, 726995
-
-
amount of tryptophanase is diminished in mutants DELTAdnaJ and DELTAdnaKdnaJ. DnaJ and DnaK molecular chaperones influence the amount of tryptophanase
-
-
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GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
metabolism
physiological function
metabolism
-
tryptophanase is an initial enzyme for the degradation of L-tryptophan and 5-hydroxytryptophan in Escherichia coli
metabolism
-
indole production requires TnaB responsible for exogenous L-tryptophan import
metabolism
-
indole production requires TnaB responsible for exogenous L-tryptophan import
-
physiological function
-
Hfq protein associates with tryptophanase under relatively higher extracellular indole levels, suggesting this is a feedback control of indole production
physiological function
-
tryptophan indole lyase produces indole from L-tryptophan, indole is a signaling molecule in bacteria, affecting biofilm formation, pathogenicity and antibiotic resistance
physiological function
-
tryptophan indole lyase produces indole from L-tryptophan, indole is a signaling molecule in bacteria, affecting biofilm formation, pathogenicity and antibiotic resistance
-
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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
D-serine
pyruvate + NH3
-
with high diammonium hydrogen phosphate concentration, e.g. 1.2 M
-
-
?
D-Trp + H2O
indole + pyruvate + NH4+
in the presence of high concentrations of ammonium phosphate
-
-
?
L-Trp + H2O
indole + pyruvate + NH3
-
effects of temperature and hydrostatic pressure on the equilibria and rate constants for quinoid intermediate formation from L-Trp and L-Met with H463 mutant enzyme
-
-
?
S-(2-nitrophenyl)-L-cysteine + H2O
2-nitrobenzenethiolate + pyruvate + NH4+
-
-
-
-
ir
S-o-nitrophenyl-L-Cys + indole
Trp + ?
-
reaction readily proceeds in water, it becomes impossible in water-organic solvents
-
-
S-o-nitrophenyl-L-cysteine + H2O
o-nitrothiophenol + pyruvate + NH4+
-
-
-
-
?
5-hydroxy-L-Trp + H2O
?
-
6.7% of the activity with L-Trp
-
-
-
5-methyl-L-Trp + H2O
?
-
15.2% of the activity with L-Trp
-
-
-
beta-(benzimidazol-1-yl)-L-alanine + H2O
?
-
reaction via aldimine intermediate, high activity with wild-type enzyme and mutant H463F
-
-
?
beta-(benzimidazol-1-yl)-L-alanine + H2O
?
-
reaction via aldimine intermediate, high activity with wild-type enzyme and mutant H463F
-
-
?
D-serine + indole
L-tryptophan + H2O
-
the presence of 20% saturation concentration of diammonium hydrogen phosphate is required for this reaction
-
-
?
D-serine + indole
L-tryptophan + H2O
-
with high diammonium hydrogen phosphate concentration, e.g. 1.2 M
-
-
?
indole + S-methyl-L-Cys
L-Trp + ?
-
beta-replacement reaction
-
-
L-Trp + H2O
indole + pyruvate + NH4+
-
-
-
-
?
L-Trp + H2O
indole + pyruvate + NH4+
-
-
-
-
r
L-Trp + H2O
indole + pyruvate + NH4+
-
the product indole is involved in cell signaling, involved in cell cycle regulation
-
-
r
L-Trp + H2O
indole + pyruvate + NH4+
-
-
-
-
L-Trp + H2O
indole + pyruvate + NH4+
-
-
-
-
L-tryptophan + H2O
indole + pyruvate + NH3
-
active to D-tryptophan in highly concentrated diammonium hydrogen phosphate solution
-
-
?
L-tryptophan + H2O
indole + pyruvate + NH3
-
reversible hydrolytic beta-elimination
-
-
r
S-(2-nitrophenyl)-L-cysteine + H2O
2-nitrophenol + pyruvate + NH4+
-
-
-
-
?
S-(2-nitrophenyl)-L-cysteine + H2O
2-nitrophenol + pyruvate + NH4+
-
-
-
?
S-(2-nitrophenyl)-L-cysteine + H2O
2-nitrophenol + pyruvate + NH4+
-
-
-
?
S-benzyl-L-Cys + H2O
phenylmethanethiol + pyruvate + NH4+
-
-
-
?
S-benzyl-L-Cys + H2O
phenylmethanethiol + pyruvate + NH4+
-
-
-
?
S-ethyl-L-Cys + H2O
ethanethiol + pyruvate + NH4+
-
-
-
?
S-ethyl-L-cysteine + H2O
ethanethiol + pyruvate + NH4+
-
-
-
?
S-methyl-L-Cys + H2O
?
-
63.8% of the activity with L-Trp
-
-
-
S-methyl-L-cysteine + H2O
methanethiol + pyruvate + NH4+
-
-
-
?
S-methyl-L-cysteine + H2O
methanethiol + pyruvate + NH4+
-
-
-
?
S-o-nitrophenyl-L-Cys + H2O
o-nitrophenol + pyruvate + NH4+
-
-
-
-
S-o-nitrophenyl-L-Cys + H2O
o-nitrophenol + pyruvate + NH4+
-
-
-
-
-
additional information
?
-
-
tryptophanase displays no activity on D-tryptophan under usual conditions. However, it becomes active toward D-tryptophan degradation in highly concentrated diammonium hydrogen phosphate solutions
-
-
-
additional information
?
-
-
synthesis of L-tryptophan from L-cysteine, pyridoxal 5'-phosphate, and indole by the recombinant enzyme expressed in Escherichia coli strain BL21(DE3)
-
-
-
additional information
?
-
-
tryptophanase does not degrade L-alanine, L-serine or L-cysteine
-
-
-
additional information
?
-
-
tryptophanase does not degrade L-alanine, L-serine or L-cysteine
-
-
-
additional information
?
-
-
the enzyme is induced by Trp and its analogs, and partially repressed by 0.5% glucose or glycerol
-
-
-
additional information
?
-
-
the enzyme is induced by Trp and its analogs, and partially repressed by 0.5% glucose or glycerol
-
-
-
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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-Trp + H2O
indole + pyruvate + NH4+
-
the product indole is involved in cell signaling, involved in cell cycle regulation
-
-
r
additional information
?
-
-
the enzyme is induced by Trp and its analogs, and partially repressed by 0.5% glucose or glycerol
-
-
-
additional information
?
-
-
the enzyme is induced by Trp and its analogs, and partially repressed by 0.5% glucose or glycerol
-
-
-
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
pyridoxal 5'-phosphate
-
binds 1 mol of pyridoxal 5'-phosphate per subunit; dependent on
pyridoxal 5'-phosphate
-
conversion of apoenzyme into the active holoenzyme is attained at 30°C in Tris-HCl buffer, pH 8.0, containing pyridoxal 5'-phosphate and K+, no conversion occurs at 5°C; dependent on
pyridoxal 5'-phosphate
-
one enzyme molecule contains 4 pyridoxal 5'-phosphate combining sites
pyridoxal 5'-phosphate
-
one enzyme molecule contains 4 pyridoxal 5'-phosphate combining sites; some coenzyme analogues can replace pyridoxal 5'-phosphate
pyridoxal 5'-phosphate
-
one enzyme molecule contains 4 pyridoxal 5'-phosphate combining sites
pyridoxal 5'-phosphate
-
one enzyme molecule contains 4 pyridoxal 5'-phosphate combining sites
pyridoxal 5'-phosphate
Escherichia aurescens
-
enzyme contain 4 mol of pyridoxal 5'-phosphate per mol of enzyme, independently of the pH in presence of K+
pyridoxal 5'-phosphate
-
enzyme contain 4 mol of pyridoxal 5'-phosphate per mol of enzyme, independently of the pH in presence of K+
pyridoxal 5'-phosphate
-
enzyme contains 4 mol of pyridoxal 5'-phosphate per mol of enzyme at pH 6.8 and 3 mol of pyridoxal 5'-phosphate per mol of enzyme at pH 7.8 in K+ buffer
pyridoxal 5'-phosphate
-
enzyme contains 4 mol of pyridoxal 5'-phosphate per mol of enzyme at pH 6.8 and 3 mol of pyridoxal 5'-phosphate per mol of enzyme at pH 7.8 in K+ buffer
pyridoxal 5'-phosphate
-
contains 4 mol of pyridoxal 5'-phosphate per mol of enzyme
pyridoxal 5'-phosphate
-
dependent on; enzyme contains 2 pyridoxal 5'-phosphate per subunit
pyridoxal 5'-phosphate
-
contains 4 mol of pyridoxal 5'-phosphate per mol of enzyme; dependent on
pyridoxal 5'-phosphate
-
contains 4 mol of pyridoxal 5'-phosphate per mol of enzyme; dependent on; Km: 0.00209 mM
pyridoxal 5'-phosphate
-
covalent binding required to activate the enzyme; dependent on
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Cl-
-
bound to enzyme, possibly required for stabilization of subunit interactions
K+
NH4+
Rb+
sulfate
-
two ions bound two the active site of the enzyme, one of the sulfate ions interacts with both the transferase and PLP-binding domains and appears to be responsible for holding the enzyme in its closed conformation
Tl+
K+
-
promotes the conversion of the inactive holoenzyme into the active holoenzyme rather than being involved in the conversion of the apoenzyme and pyridoxal 5'-phosphate into the active holoenzyme
K+
-
kinetics of tryptophanase inactivation/activation by sudden removal/addition of K+ with the aid of a crown ether or cryptand
K+
-
stabilizing, cold inactivation occurs more slowly in the presence of K+
NH4+
-
promotes the conversion of the inactive holoenzyme into the active holoenzyme rather than being involved in the conversion of the apoenzyme and pyridoxal 5'-phosphate into the active holoenzyme
NH4+
-
monovalent cation required for activity and for tight cofactor binding
Tl+
-
monovalent cation required for activity and for tight cofactor binding
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INHIBITORS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
2-amino-4-(benzimidazol-1-yl)butyric acid
-
i.e. homo-BZI-Ala, a potent competitive inhibitor
2-amino-5-(benzimidazol-1-yl)pentanoic acid
-
i.e. bishomo-BZI-Ala, weak inhibition
alpha-amino-9,10-dihydro-9,10-dioxo-2-anthracenepropanoic acid
-
noncompetitive inhibition
beta-(benzimidazol-1-yl)-L-alanine
-
competitive, is also a good substrate for the wild-type and mutant enzymes
Bifidobacterium adolescentis SPM0212
-
inhibits the proliferation of human colon cancer cell lines and it inhibits harmful fecal enzymes of rat intestinal microflora, including alpha-glucuronidase, alpha-glucosidase, tryptophanase, and urease
-
cyclodextrin
-
mixed type inhibition, competitive and non-competitive with inhibitor constants for different cyclodextrins between 0.5 and 10 mM
-
diammonium hydrogen phosphate
-
diammonium hydrogen phosphate serves as an inhibitor of tryptophanase when L-serine is substrate, the activity decreases with increasing diammonium hydrogen phosphate
additional information
-
cyclodextrin derivatives cause the inhibition of enzymatic process, both competitive and non-competitive. The competitive inhibition is connected with the formation of inclusion complexes between cyclodextrins and L-tryptophan, related to the geometry of these complexes. The mechanism of the non-competitive inhibition is not so evident
-
additional information
-
when bound to silver nanoparticles, TNase activity decreases significantly by 50% for carbonate coated silver nanoparticles and by over 90% for bare silver nanoparticles
-
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
diammonium hydrogen phosphate
-
diammonium hydrogen phosphate serves as an activator on tryptophan synthesis from D-serine, maximum activity at 20% saturation concentration
sRNA
-
dimers of plasmid ColE1 make an sRNA that interacts directly with the enzyme and enhance its substrate affinity
-
additional information
-
NaCl, NH4H2PO4, NH4NO3, NH4Cl, (BH4)2CO3, K2HPO4, and Na2HPO4 have no effect on enzyme activity
-
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.09
L-Trp
-
purified recombinant tryptophanase in 200 mM sodium phosphate buffer (pH 7.5), at 37°C
0.1732
L-Trp
-
in the presence of Hfq protein, in 0.1 M potassium phosphate buffer (pH 7.8), at 37°C
0.1901
L-Trp
-
in the absence of Hfq protein, in 0.1 M potassium phosphate buffer (pH 7.8), at 37°C
0.2
L-Trp
-
purified recombinant tryptophanase in 200 mM potassium phosphate buffer (pH 7.5), at 37°C
0.26
S-ethyl-L-cysteine
-
purified recombinant tryptophanase in 200 mM potassium phosphate buffer (pH 7.5), at 37°C
2.04
S-methyl-L-cysteine
-
purified recombinant tryptophanase in 200 mM potassium phosphate buffer (pH 7.5), at 37°C
additional information
additional information
-
kinetics of mutant H463F, high-pressure stopped-flow measurements, overview
-
additional information
additional information
-
pre-steady-state and steady-state kinetics of wild-type and mutant enzymes, overview
-
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.24
L-Trp
-
purified recombinant tryptophanase in 200 mM sodium phosphate buffer (pH 7.5), at 37°C
1.37
L-Trp
-
purified recombinant tryptophanase in 200 mM potassium phosphate buffer (pH 7.5), at 37°C
1.87
S-ethyl-L-cysteine
-
purified recombinant tryptophanase in 200 mM potassium phosphate buffer (pH 7.5), at 37°C
0.5
S-methyl-L-cysteine
-
purified recombinant tryptophanase in 200 mM potassium phosphate buffer (pH 7.5), at 37°C
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
6.87
L-Trp
-
purified recombinant tryptophanase in 200 mM potassium phosphate buffer (pH 7.5), at 37°C
7.15
S-ethyl-L-cysteine
-
purified recombinant tryptophanase in 200 mM potassium phosphate buffer (pH 7.5), at 37°C
0.25
S-methyl-L-cysteine
-
purified recombinant tryptophanase in 200 mM potassium phosphate buffer (pH 7.5), at 37°C
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Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0134
2-amino-4-(benzimidazol-1-yl)butyric acid
-
wild-type enzyme, pH 8.0, 25°C
0.6
2-amino-5-(benzimidazol-1-yl)pentanoic acid
-
above, wild-type enzyme, pH 8.0, 25°C
174
alpha-amino-9,10-dihydro-9,10-dioxo-2-anthracenepropanoic acid
-
in 50 mM potassium phosphate buffer (pH 7.8), at 25°C
52
L-tryptophan ethylester
-
in 50 mM potassium phosphate buffer (pH 7.8), at 25°C
48
N-acetyl-L-tryptophan
-
in 50 mM potassium phosphate buffer (pH 7.8), at 25°C
0.005
oxindolyl-L-alanine
-
in 50 mM potassium phosphate buffer (pH 7.8), at 25°C
101
S-phenylbenzoquinone-L-tryptophan
-
in 50 mM potassium phosphate buffer (pH 7.8), at 25°C
-
additional information
additional information
-
Ki-values for reaction in aqueous methanol
-
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.508
-
cell extract from strain MG1655H, harvested at OD600 of 0.8, at 37°C
0.808
-
cell extract from strain MG1655H, harvested at OD600 of 0.4, at 37°C
additional information
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7 - 9.8
-
pH 7: about 45% of maximal activity, pH 9.8: about 90% of maximal activity
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
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PDB
SCOP
CATH
ORGANISM
UNIPROT
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
50000
52000
52500
55000
210000
216000
-
aggregation to form species with MW 432000 Da, 648000 Da, and 864000 Da, sucrose density gradient centrifugation, equilibrium sedimentation
220000
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SUBUNITS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
dimer
homodimer
tetramer
additional information
tetramer
-
4 * 57000, equilibrium sedimentation in 6 M guanidine hydrochloride
additional information
-
upon cooling to 2°C, inactivation and dissociation of tetramer to dimer occurs, spectrofluorometry data
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Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
apo-form of enzyme without bound pyridoxal 5-phosphate but with two bound sulfate ions, hanging drop vapor diffusion method; strong crystal contacts occur on the flat surface of the protein and that the size of crystal contact surface seems to correlate with the diffraction quality of the crystal. The tryptophanase structure, solved in its apo form, does not have covalent PLP bound in the active site, but two sulfate ions. The sulfate ions occupy the phosphoryl-binding site of PLP and the binding site of the alpha-carboxyl of the natural substrate tryptophan. One of the sulfate ions makes extensive interactions with both the transferase and PLP-binding domains of the protein and appears to be responsible for holding the enzyme in its closed conformation
-
crystallized in the apo form by the hanging-drop vapour-diffusion method using polyethylene glycol 400 as a precipitant and magnesium chloride as an additive. The crystals belong to the orthorhombic space group F222, with unit-cell parameters a = 118.4 A, b = 120.1 A, c = 171.2 A. Contains a monomer in the asymmetric unit with a solvent content of 55%. Tryptophanase mutants W330F and Y74F are crystallized under the same conditions and the crystals diffracted to a resolution limit of 1.9 A
-
mutant enzymes Y74F and C298S, hanging drop vapor diffusion method, using 30% (w/v) PEG 400, 100 mM HEPES pH 7.5, 200 mM MgCl2, 5 mM 2-mercaptoethanol
-
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pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
40 - 70
-
the enzyme is stable up to 40°C, but only 40% of the activity remains at 60°C, the enzyme is completely inactivated at temperatures above 70°C
55
60
65
80
-
5 min, Tna1 retains alomost complete activity, Tna2 is inactivated
90
-
5 min, Tna1 retains more than 70% of activity, Tna2 is inactivated
additional information
2
-
upon cooling to 2°C, inactivation and dissociation of teramer to dimer occurs, spectrofluorometry data
2
-
subsequent to incubation at 2°C, wild type Trpase loses about 35% of its activity followed by dissociation into dimers. Complete dissociation/inactivation occurs after incubation for 20 h at 2°C and association/reactivation occurs upon warming for few hours
55
-
10 min, apoenzyme, about 90% loss of activity; 10 min, holoenzyme, stable
additional information
-
mutant strain ts6 with an enzyme with higher thermostability
additional information
-
inactivation of the enzyme at low temperatures due to dissociation of the active tetramer into dimers upon cold incubation
additional information
-
about 80% activity at 50°C after 2 hours, about 20% activity after incubation at 0°C for 3 hours
additional information
-
rapid loss of activity at 50°C, no change in activity on incubation at 0°C for more than 3 hours
additional information
rapid loss of activity at 50°C, no change in activity on incubation at 0°C for more than 3 hours
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GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
activity decreases at pressures above 50 MPa, and by 100 MPa is less than 10% of the activity at 1 bar, initial increase in activity with pressure, reaching a maximum of about 140% at 30 MPa.
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greater than 90% activity remaining at 100 MPa, initial increase in activity with pressure, reaching a maximum of about 140% at 60 MPa.
immobilized enzyme shows higher thermal stability and resistance to a denaturing agent such as guanidine-HCl than the soluble enzyme
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rapid inactivation by visible light irradiation in presence of pyridoxal 5'-phosphate. Photoinactivation follows pseudo-first-order kinetics
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repeated slow freezing at -20°C and subsequent thawing at room temperature causes the enzyme to aggregate
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when used repeatedly in a batch system or continously in a flow system in the absence of added pyridoxal 5'-phosphate, immobilized holo-tryptophanase gradually loses its original activity, pyridoxal 5'-phosphate restores its initial activity
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, 8-10 mg/ml in 0.1 M potassium phosphate buffer, pH 7.8, containing 0.1 mM pyridoxal phosphate and 1.0 mM dithiothreitol, stable for up to 1 year
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-70°C, stored in 0.1 M potassium phosphate buffer, pH 6.8
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5°C, stored as a suspension in 0.1 M potassium phosphate buffer, pH 8.0, containing 20% v/v glycerol, 0.1 mM pyridoxal 5'-phosphate, 10 mM mercaptoethanol and 60% saturated ammonium sulfate, stable for more than 5 months
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stored as a suspension under nitrogen in 0.1 M potassium phosphate, pH 8.0, 0.2 mM dithiothreitol, 1 mM EDTA, and 2 M (NH4)2SO4
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DEAE-Sephadex A-50 gel filtration or DEAE-Sepharose 4B column chromatography, and Sephacryl S-300 gel filtration
-
no added pyridoxal 5-phosphate during purification procedure resulting in the inactive apo-form of the enzyme
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in wine yeast strains Saccharomyces cerevisiae strains YHUM272a and VIN13, expression of enzyme results in a strong increase of passion fruit aroma in wine
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
the endogenous oxidative stress in ibpAB cells leads to increased expression of tryptophanase
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tryptophanase activity is decreased by 43% after lactic acid bacteria treatment (300 billion CFU/g twice a day for 2 weeks)
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tryptophanase is upregulated in the noninvasive ibeR deletion mutant BR2
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
C298S
-
the mutant displays reduced activity, subsequent to incubation at 2°C, the mutant Trpase loses about 90% of its activity
H463F
W330F
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
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
-
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
-
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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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)
additional information
H463F
-
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, pH dependence of quinonoid intermediate formation, overview
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
W330F
-
as in wild type, upon cooling to 2°C, inactivation and dissociation of tetramer to dimer occurs, spectrofluorometry data
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
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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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
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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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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
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Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
biotechnology
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encapsulation of enzyme in wet nanoporous silica gels to selectively stabilize tertiary and quarternary protein conformations and to develop bioreactors and biosensors
food industry
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expression of enzyme in wine yeast results in a strong increase of passion fruit aroma in wine
medicine
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transposon insertion in gene results in a strain unable to form biofilm on polystyrene and to adhere to human pneumocyte cells
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LINKS TO OTHER DATABASES (specific for EC-Number 4.1.99.1)
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