Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Nuidate, T.; Tansila, N.; Chomchuen, P.; Phattaranit, P.; Eangchuan, S.; Vuddhakul, V.
    Characterization of tryptophanase from Vibrio cholerae (2015), Appl. Biochem. Biotechnol., 175, 243-252 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.1.99.1 gene tnaA, recombinant expression of His6-tagged enzyme VcTrpase in Escherichia coli strain BL21(DE3) tn5:tnaA Vibrio cholerae O1

Protein Variants

EC Number Protein Variants Comment Organism
4.1.99.1 D137A site-directed mutagenesis, inactive mutant Vibrio cholerae O1
4.1.99.1 H463A site-directed mutagenesis, the mutant shows 96% reduced activity compared to the wild-type Vibrio cholerae O1
4.1.99.1 K269A site-directed mutagenesis, inactive mutant Vibrio cholerae O1
4.1.99.1 K270A site-directed mutagenesis, the mutation possibly eliminates this cofactor-protein interaction in the enzyme, inactive mutant Vibrio cholerae O1
4.1.99.1 R103A site-directed mutagenesis, the mutation alters orientation of the cofactor pyridoxal 5'-phosphate, inactive mutant Vibrio cholerae O1
4.1.99.1 R230A site-directed mutagenesis, inactive mutant Vibrio cholerae O1
4.1.99.1 Y52A site-directed mutagenesis, inactive mutant Vibrio cholerae O1
4.1.99.1 Y74A site-directed mutagenesis, inactive mutant Vibrio cholerae O1

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.1.99.1 additional information
-
 additional information Michaelis-Menten kinetics Vibrio cholerae O1
4.1.99.1 0.346
-
 S-Benzyl-L-cysteine recombinant His-tagged wild-type enzyme, pH 7.8, 37°C Vibrio cholerae O1
4.1.99.1 0.49
-
 S-methyl-L-cysteine recombinant His-tagged wild-type enzyme, pH 7.8, 37°C Vibrio cholerae O1
4.1.99.1 5.252
-
 L-tryptophan recombinant His-tagged wild-type enzyme, pH 7.8, 37°C Vibrio cholerae O1

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4.1.99.1 L-tryptophan + H2O Vibrio cholerae O1
-
 indole + pyruvate + NH3
-
 ?
4.1.99.1 L-tryptophan + H2O Vibrio cholerae O1 ATCC 39541
-
 indole + pyruvate + NH3
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
4.1.99.1 Vibrio cholerae O1 A5EYI0
-
-
4.1.99.1 Vibrio cholerae O1 ATCC 39541 A5EYI0
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.1.99.1 recombinant His6-tagged enzyme VcTrpase from Escherichia coli strain BL21(DE3) tn5:tnaA by nickel affinity chromatography Vibrio cholerae O1

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.1.99.1 L-tryptophan + H2O
-
 Vibrio cholerae O1 indole + pyruvate + NH3
-
 ?
4.1.99.1 L-tryptophan + H2O
-
 Vibrio cholerae O1 ATCC 39541 indole + pyruvate + NH3
-
 ?
4.1.99.1 additional information coupled assay: the decrease in NADH absorbance at 340 nm in the presence of LDH is used to determine Trpase activity after adding the substrate. L-Phenylalanine and L-serine are no substrates Vibrio cholerae O1 ?
-
 ?
4.1.99.1 additional information coupled assay: the decrease in NADH absorbance at 340 nm in the presence of LDH is used to determine Trpase activity after adding the substrate. L-Phenylalanine and L-serine are no substrates Vibrio cholerae O1 ATCC 39541 ?
-
 ?
4.1.99.1 S-benzyl-L-cysteine + H2O
-
 Vibrio cholerae O1 phenylmethanethiol + pyruvate + NH3
-
 ?
4.1.99.1 S-benzyl-L-cysteine + H2O
-
 Vibrio cholerae O1 ATCC 39541 phenylmethanethiol + pyruvate + NH3
-
 ?
4.1.99.1 S-methyl-L-cysteine + H2O
-
 Vibrio cholerae O1 methanethiol + pyruvate + NH3
-
 ?
4.1.99.1 S-methyl-L-cysteine + H2O
-
 Vibrio cholerae O1 ATCC 39541 methanethiol + pyruvate + NH3
-
 ?

Subunits

EC Number Subunits Comment Organism
4.1.99.1 ? x * 49000, His6-tagged enzyme VcTrpase, SDS-PAGE Vibrio cholerae O1

Synonyms

EC Number Synonyms Comment Organism
4.1.99.1 TnaA
-
 Vibrio cholerae O1
4.1.99.1 Trpase
-
 Vibrio cholerae O1
4.1.99.1 VcTrpase
-
 Vibrio cholerae O1

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.1.99.1 45
-
-
 Vibrio cholerae O1

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
4.1.99.1 22 70 activity range, inactivation above Vibrio cholerae O1

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
4.1.99.1 4
-
 VcTrpase does not dissociate into inactive dimers or monomers at 4°C Vibrio cholerae O1

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.1.99.1 0.346
-
 S-Benzyl-L-cysteine recombinant His-tagged wild-type enzyme, pH 7.8, 37°C Vibrio cholerae O1
4.1.99.1 0.49
-
 S-methyl-L-cysteine recombinant His-tagged wild-type enzyme, pH 7.8, 37°C Vibrio cholerae O1
4.1.99.1 5.252
-
 L-tryptophan recombinant His-tagged wild-type enzyme, pH 7.8, 37°C Vibrio cholerae O1

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
4.1.99.1 9
-
-
 Vibrio cholerae O1

pH Range

EC Number pH Minimum pH Maximum Comment Organism
4.1.99.1 6 9 high activity within this range Vibrio cholerae O1

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
4.1.99.1 6.5 8 purified recombinant His-tagged enzyme, 10 min, over 80% activity remaining Vibrio cholerae O1

Cofactor

EC Number Cofactor Comment Organism Structure
4.1.99.1 pyridoxal 5'-phosphate dependent on, residue Arg103 plays an important role in orientation of the cofactor pyridoxal 5'-phosphate Vibrio cholerae O1

General Information

EC Number General Information Comment Organism
4.1.99.1 additional information eight catalytically important residues Thr52, Tyr74, Arg103, Asp137, Arg230, Lys269, Lys270, and His463 are located close to the Trpase active site and are absolutely conserved in Trpases. Five of them are located in the conserved regions and are reported to confer a crucial role for binding of the substrate and cofactor to produce the formation of the best intermediate that will lead to substrate degradation. Despite the apparent diversity in the protein sequences, these regions may be essential for enzyme activity to generate indole, which is an important agent for bacterial physiology, ecological balance, and virulence Vibrio cholerae O1
4.1.99.1 physiological function the enzyme is responsible for the production of indole, an important intra- and interspecies signaling molecule in bacteria Vibrio cholerae O1

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
4.1.99.1 0.016
-
 S-methyl-L-cysteine recombinant His-tagged wild-type enzyme, pH 7.8, 37°C Vibrio cholerae O1
4.1.99.1 1.648
-
 S-Benzyl-L-cysteine recombinant His-tagged wild-type enzyme, pH 7.8, 37°C Vibrio cholerae O1
4.1.99.1 8.582
-
 L-tryptophan recombinant His-tagged wild-type enzyme, pH 7.8, 37°C Vibrio cholerae O1