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Literature summary extracted from

  • Hettwer, S.; Sterner, R.
    A novel tryptophan synthase beta-subunit from the hyperthermophile Thermotoga maritima. Quaternary structure, steady-state kinetics, and putative physiological role (2002), J. Biol. Chem., 277, 8194-8201.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.2.1.20 genes trpA, trpB1, and trpB2, expression in Escherichia coli Thermotoga maritima
4.2.1.122 expressed in Escherichia coli BL21(DE3) cells Thermotoga maritima

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.2.1.20 additional information
-
 additional information steady state kinetics Thermotoga maritima
4.2.1.20 0.025
-
 indole pH 7.5, 80°C, recombinant (TrpA-TrpB1)2 tetrameric complex, beta-reaction Thermotoga maritima
4.2.1.20 0.04
-
 indole pH 7.5, 80°C, recombinant TrpB1, beta-reaction Thermotoga maritima
4.2.1.20 0.19
-
 1-(indol-3-yl)glycerol 3-phosphate pH 7.5, 80°C, recombinant (TrpA-TrpB1)2 tetrameric complex, alpha-reaction Thermotoga maritima
4.2.1.20 1.62
-
 1-(indol-3-yl)glycerol 3-phosphate pH 7.5, 80°C, recombinant TrpA, alpha-reaction Thermotoga maritima
4.2.1.20 3.7
-
 L-serine pH 7.5, 80°C, recombinant (TrpA-TrpB1)2 tetrameric complex, beta-reaction Thermotoga maritima
4.2.1.20 110
-
 L-serine pH 7.5, 80°C, recombinant TrpB1, beta-reaction Thermotoga maritima
4.2.1.122 0.00077
-
 indole Km below 0.00077 mM, at 80°C, pH not specified in the publication Thermotoga maritima
4.2.1.122 50.2
-
 L-serine at 80°C, pH not specified in the publication Thermotoga maritima

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
4.2.1.20 26200
-
 recombinant TrpA monomer, sedimentation equilibrium Thermotoga maritima
4.2.1.20 91000
-
 recombinant TrpB1 dimer, sedimentation equilibrium Thermotoga maritima
4.2.1.20 98000
-
 recombinant TrpB2 dimer, sedimentation equilibrium Thermotoga maritima
4.2.1.20 140000
-
 recombinant alphabetabetaalpha complex of TRpA and TrpB1, sedimentation equilibrium Thermotoga maritima
4.2.1.122 46400
-
 2 * 46400, calculated from amino acid sequence Thermotoga maritima
4.2.1.122 61700
-
 gel filtration Thermotoga maritima
4.2.1.122 92770
-
 calculated from amino acid sequence Thermotoga maritima
4.2.1.122 98000
-
 sedimentation equilibrium analysis Thermotoga maritima

Organism

EC Number Organism UniProt Comment Textmining
4.2.1.20 Thermotoga maritima
-
 hyperthermophile, alpha-subunit TrpA and beta-subunit TrpB1 are encoded by the 2 genes trpA and trpB1 in the trp operon, and by a trpB2 gene outside the operon
-
4.2.1.122 Thermotoga maritima
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.2.1.20 recombinant TrpA, TrpB1, and TrpB2 from Escherichia coli Thermotoga maritima
4.2.1.122 Mono Q column chromatography and Superdex 75 gel filtration Thermotoga maritima

Reaction

EC Number Reaction Comment Organism Reaction ID
4.2.1.20 L-serine + 1-C-(indol-3-yl)glycerol 3-phosphate = L-tryptophan + D-glyceraldehyde 3-phosphate + H2O also catalyses the conversion of serine and indole into tryptophan and water, and of indoleglycerol phosphate into indole and glyceraldehyde phosphate (the latter reaction was listed formerly as EC 4.2.1.8) Thermotoga maritima
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.2.1.20 1-(indol-3-yl)glycerol 3-phosphate alpha-subunit of the bienzyme complex, alpha-reaction Thermotoga maritima D-glyceraldehyde 3-phosphate + indole
-
 ?
4.2.1.20 L-serine + indole beta-subunit of the bienzyme complex, beta-reaction Thermotoga maritima L-tryptophan + H2O
-
 ?
4.2.1.20 additional information indole is channeled to the active site of the beta-subunit, the TrpB2 homodimer has a high catalytic efficiency due to a low Km for indole, it functions as a rescue protein for indole to prevent the escape of the costly hydrophobic metabolite at the hig growing temperatures of the hyperthermophile Thermotoga maritima ?
-
 ?
4.2.1.122 L-serine + indole
-
 Thermotoga maritima L-tryptophan + H2O
-
 ?
4.2.1.122 additional information TrpB2 does not bind to TrpA but is catalytically highly active, has an extremely low Km value for indole Thermotoga maritima ?
-
 ?

Subunits

EC Number Subunits Comment Organism
4.2.1.20 dimer TrpB2 does not bind to the alpha-subunit and forms homodimers, TrpB1 alone forms homodimers Thermotoga maritima
4.2.1.20 monomer TrpA alone forms monomers Thermotoga maritima
4.2.1.20 More recombinantly expressed alpha-subunit forms monomers of 26.7 kDa, while the recombinant beta-subunits B1 and B2 both form dimers of 49.4 and 61.7 kDa, respectively Thermotoga maritima
4.2.1.20 tetramer alphabetabetaalpha complex of TRpA and TrpB1, sedimentation equilibrium Thermotoga maritima
4.2.1.122 homodimer 2 * 46400, calculated from amino acid sequence Thermotoga maritima

Synonyms

EC Number Synonyms Comment Organism
4.2.1.20 TrpA
-
 Thermotoga maritima
4.2.1.20 TrpB1
-
 Thermotoga maritima
4.2.1.20 TrpB2
-
 Thermotoga maritima
4.2.1.122 TrpB2
-
 Thermotoga maritima

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.2.1.20 30 60 alpha-reaction assay at Thermotoga maritima
4.2.1.20 80
-
 beta-reaction assay at Thermotoga maritima

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.2.1.20 0.174
-
 1-(indol-3-yl)glycerol 3-phosphate pH 7.5, 80°C, recombinant TrpA, alpha-reaction Thermotoga maritima
4.2.1.20 3.8
-
 L-serine pH 7.5, 80°C, recombinant TrpB1, beta-reaction Thermotoga maritima
4.2.1.20 4.2
-
 indole pH 7.5, 80°C, recombinant TrpB1, beta-reaction Thermotoga maritima
4.2.1.20 5.6
-
 1-(indol-3-yl)glycerol 3-phosphate pH 7.5, 80°C, recombinant (TrpA-TrpB1)2 tetrameric complex, alpha-reaction Thermotoga maritima
4.2.1.20 8
-
 L-serine pH 7.5, 80°C, recombinant (TrpA-TrpB1)2 tetrameric complex, beta-reaction Thermotoga maritima
4.2.1.20 10
-
 indole pH 7.5, 80°C, recombinant (TrpA-TrpB1)2 tetrameric complex, beta-reaction Thermotoga maritima
4.2.1.122 0.44
-
 L-serine at 80°C, pH not specified in the publication Thermotoga maritima
4.2.1.122 0.46
-
 indole at 80°C, pH not specified in the publication Thermotoga maritima

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
4.2.1.20 7.5
-
 assay at Thermotoga maritima

General Information

EC Number General Information Comment Organism
4.2.1.122 physiological function TrpB2 acts as an indole rescue protein, which prevents the escape of this costly hydrophobic metabolite from the cell at the high growth temperatures of hyperthermophiles Thermotoga maritima

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
4.2.1.122 0.0006
-
 indole KCAT_KM below 0.0006 mM, at 80°C, pH not specified in the publication Thermotoga maritima
4.2.1.122 8700
-
 L-serine at 80°C, pH not specified in the publication Thermotoga maritima