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Literature summary extracted from

  • Hiyama, T.; Sato, T.; Imanaka, T.; Atomi, H.
    The tryptophan synthase beta-subunit paralogs TrpB1 and TrpB2 in Thermococcus kodakarensis are both involved in tryptophan biosynthesis and indole salvage (2014), FEBS J., 281, 3113-3125.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.2.1.20 expressed in Escherichia coli BL21 cells Thermococcus kodakarensis
4.2.1.20 expression in Escherichia coli Thermococcus kodakarensis
4.2.1.122 gene trpB1, phylogenetic tree, expression in Escherichia coli strain BL21, transcript levels of trpB1 in the host, DELTAtrpB1 strains by quantitative RT-PCR expression analysis Thermococcus kodakarensis
4.2.1.122 gene trpB2, phylogenetic tree, expression in Escherichia coli strain BL21, transcript levels of trpB2 in the host, DELTAtrpB2 strains by quantitative RT-PCR expression analysis Thermococcus kodakarensis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.2.1.20 0.0076
-
 indole pH 7.5, 85°C, reaction catalyzed by TrpB2 in presence of subunit TrpA Thermococcus kodakarensis
4.2.1.20 0.0076
-
 L-serine subunit TrpB2 plus subunit TrpA, at pH 7.5 and 85°C Thermococcus kodakarensis
4.2.1.20 0.0081
-
 indole pH 7.5, 85°C, reaction catalyzed by TrpB1 Thermococcus kodakarensis
4.2.1.20 0.0081
-
 L-serine subunit TrpB2, at pH 7.5 and 85°C Thermococcus kodakarensis
4.2.1.20 0.035
-
 indole pH 7.5, 85°C, reaction catalyzed by TrpB1 in presence of subunit TrpA Thermococcus kodakarensis
4.2.1.20 0.035
-
 L-serine subunit TrpB1 plus subunit TrpA, at pH 7.5 and 85°C Thermococcus kodakarensis
4.2.1.20 0.0629
-
 indole pH 7.5, 85°C, reaction catalyzed by TrpB2 Thermococcus kodakarensis
4.2.1.20 0.629
-
 L-serine subunit TrpB1, at pH 7.5 and 85°C Thermococcus kodakarensis
4.2.1.122 additional information
-
 additional information steady-state Michaelis-Menten kinetics, overview Thermococcus kodakarensis
4.2.1.122 0.0076
-
 indole pH 7.5, 85°C, recombinant TrpB2, with TrpA Thermococcus kodakarensis
4.2.1.122 0.0081
-
 indole pH 7.5, 85°C, recombinant TrpB2, without TrpA Thermococcus kodakarensis
4.2.1.122 0.035
-
 indole pH 7.5, 85°C, recombinant TrpB1 with TrpA Thermococcus kodakarensis
4.2.1.122 0.063
-
 indole pH 7.5, 85°C, recombinant TrpB1 Thermococcus kodakarensis

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
4.2.1.20 20800
-
 2 * 20800 + 2 * 67400, heterotetramer from subunits TrpA and TrpB1, SDS-PAGE Thermococcus kodakarensis
4.2.1.20 27700
-
 2 * 27700 (subunit TrpA) + 2 * 42600 (subunit TrpB1), calculated from sequence, tryptophan synthase is composed of subunit TrpA and subunit TrpB. Thermococcus kodakarensis possess two paralogs of subunit B, TrpB1 and TrpB2. TrpB2 catalyzes the TrpB reaction but does not interact with TrpA as in the case of TrpB1 Thermococcus kodakarensis
4.2.1.20 27700
-
 2 * 27700 (subunit TrpA) + 2 * 49300 (subunit TrpB2), calculated from sequence, tryptophan synthase is composed of subunit TrpA and subunit TrpB. Thermococcus kodakarensis possess two paralogs of subunit B, TrpB1 and TrpB2. TrpB2 catalyzes the TrpB reaction but does not interact with TrpA as in the case of TrpB1 Thermococcus kodakarensis
4.2.1.20 27700
-
 2 * 27700 + 2 * 42600, heterotetramer from subunits TrpA and TrpB1, calculated from amino acid sequence Thermococcus kodakarensis
4.2.1.20 42600
-
 2 * 27700 (subunit TrpA) + 2 * 42600 (subunit TrpB1), calculated from sequence, tryptophan synthase is composed of subunit TrpA and subunit TrpB. Thermococcus kodakarensis possess two paralogs of subunit B, TrpB1 and TrpB2. TrpB2 catalyzes the TrpB reaction but does not interact with TrpA as in the case of TrpB1 Thermococcus kodakarensis
4.2.1.20 42600
-
 2 * 27700 + 2 * 42600, heterotetramer from subunits TrpA and TrpB1, calculated from amino acid sequence Thermococcus kodakarensis
4.2.1.20 49300
-
 2 * 27700 (subunit TrpA) + 2 * 49300 (subunit TrpB2), calculated from sequence, tryptophan synthase is composed of subunit TrpA and subunit TrpB. Thermococcus kodakarensis possess two paralogs of subunit B, TrpB1 and TrpB2. TrpB2 catalyzes the TrpB reaction but does not interact with TrpA as in the case of TrpB1 Thermococcus kodakarensis
4.2.1.20 67400
-
 2 * 20800 + 2 * 67400, heterotetramer from subunits TrpA and TrpB1, SDS-PAGE Thermococcus kodakarensis
4.2.1.20 140600
-
 heterotetramer from subunits TrpA and TrpB1, calculated from amino acid sequence Thermococcus kodakarensis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4.2.1.20 indoleglycerol phosphate Thermococcus kodakarensis the reaction is catalyzed by the tryptophan synthase alpha-subunit TrpA indole + D-glyceraldehyde 3-phosphate
-
 r
4.2.1.20 L-serine + 1-C-(indol-3-yl)glycerol 3-phosphate Thermococcus kodakarensis overall reaction L-tryptophan + D-glyceraldehyde 3-phosphate + H2O
-
 ir
4.2.1.20 L-serine + 1-C-(indol-3-yl)glycerol 3-phosphate Thermococcus kodakarensis the last two steps of L-tryptophan biosynthesis are catalyzed by Trp synthase, a heterotetramer composed of TrpA and TrpB. TrpB catalyzes the condensation of indole, synthesized by TrpA, and serine to Trp L-tryptophan + D-glyceraldehyde 3-phosphate + H2O
-
 ?
4.2.1.20 L-serine + indole Thermococcus kodakarensis the reaction is catalyzed by the tryptophan synthase beta-subunit paralogs TrpB1 and TrpB2 L-tryptophan + H2O
-
 ir
4.2.1.122 L-serine + indole Thermococcus kodakarensis
-
 L-tryptophan + H2O
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
4.2.1.20 Thermococcus kodakarensis
-
-
-
4.2.1.20 Thermococcus kodakarensis Q9YGA9 and Q5JDJ1 Q9YGA9: alpha-subunit, Q5JDJ1: beta-subunit TrpB2
-
4.2.1.20 Thermococcus kodakarensis Q9YGA9 and Q9YGB0 Q9YGA9: alpha-subunit, Q9YGB0: beta-subunit TrpB1
-
4.2.1.122 Thermococcus kodakarensis Q5JDJ1 TrpB2; gene trpB2, located outside the trpCDEGFB1A operon
-
4.2.1.122 Thermococcus kodakarensis Q9YGB0 TrpB1; gene trpB1, located adjacently to gene trpA in the trpCDEGFB1A operon
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.2.1.20
-
 Thermococcus kodakarensis
4.2.1.20 Resource 15 ISO column chromatography, Resource Q column chromatography, and Superdex 200 gel filtration Thermococcus kodakarensis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.2.1.20 indoleglycerol phosphate the reaction is catalyzed by the tryptophan synthase alpha-subunit TrpA Thermococcus kodakarensis indole + D-glyceraldehyde 3-phosphate
-
 r
4.2.1.20 L-serine + 1-C-(indol-3-yl)glycerol 3-phosphate overall reaction Thermococcus kodakarensis L-tryptophan + D-glyceraldehyde 3-phosphate + H2O
-
 ir
4.2.1.20 L-serine + 1-C-(indol-3-yl)glycerol 3-phosphate the last two steps of L-tryptophan biosynthesis are catalyzed by Trp synthase, a heterotetramer composed of TrpA and TrpB. TrpB catalyzes the condensation of indole, synthesized by TrpA, and serine to Trp Thermococcus kodakarensis L-tryptophan + D-glyceraldehyde 3-phosphate + H2O
-
 ?
4.2.1.20 L-serine + 1-C-(indol-3-yl)glycerol 3-phosphate subunit TrpB catalyzes the condensation of indole, synthesized by subunit TrpA, and serine to Trp Thermococcus kodakarensis L-tryptophan + D-glyceraldehyde 3-phosphate + H2O
-
 ?
4.2.1.20 L-serine + indole reaction is catalyzed by the beta-subunit, TrpB1 or TRpB2. TrpB1 is activated by the addition of the alpha-subunit TrpA and subsequent complex formation, whereas the activity of TrpB2 is not significantly affected by TrpA Thermococcus kodakarensis L-tryptophan + H2O
-
 ?
4.2.1.20 L-serine + indole the reaction is catalyzed by the tryptophan synthase beta-subunit paralogs TrpB1 and TrpB2 Thermococcus kodakarensis L-tryptophan + H2O
-
 ir
4.2.1.20 additional information both subunit TrpB1 and TrpB2 show trace levels of serine deaminase activity by converting L-serine to pyruvate and ammonia Thermococcus kodakarensis ?
-
 ?
4.2.1.122 L-serine + indole
-
 Thermococcus kodakarensis L-tryptophan + H2O
-
 ?

Subunits

EC Number Subunits Comment Organism
4.2.1.20 heterotetramer 2 * 27700 (subunit TrpA) + 2 * 42600 (subunit TrpB1), calculated from sequence, tryptophan synthase is composed of subunit TrpA and subunit TrpB. Thermococcus kodakarensis possess two paralogs of subunit B, TrpB1 and TrpB2. TrpB2 catalyzes the TrpB reaction but does not interact with TrpA as in the case of TrpB1 Thermococcus kodakarensis
4.2.1.20 heterotetramer 2 * 27700 (subunit TrpA) + 2 * 49300 (subunit TrpB2), calculated from sequence, tryptophan synthase is composed of subunit TrpA and subunit TrpB. Thermococcus kodakarensis possess two paralogs of subunit B, TrpB1 and TrpB2. TrpB2 catalyzes the TrpB reaction but does not interact with TrpA as in the case of TrpB1 Thermococcus kodakarensis
4.2.1.20 heterotetramer 2 * 20800 + 2 * 67400, heterotetramer from subunits TrpA and TrpB1, SDS-PAGE Thermococcus kodakarensis
4.2.1.20 heterotetramer 2 * 27700 + 2 * 42600, heterotetramer from subunits TrpA and TrpB1, calculated from amino acid sequence Thermococcus kodakarensis
4.2.1.122 heterotetramer Trp synthase is a heterotetramer composed of TrpA and TrpB, TrpA and TrpB1 assemble to form an alphabetabetaalpha heterotetramer Thermococcus kodakarensis
4.2.1.122 More complex formation of TrpB1 and TrpB2 with TrpA is analyzed by gel filtration Thermococcus kodakarensis
4.2.1.122 More complex formation of TrpB1 and TrpB2 with TrpA is analyzed by gel filtration, TrpA and TrpB2 do not form a complex Thermococcus kodakarensis

Synonyms

EC Number Synonyms Comment Organism
4.2.1.20 Trp synthase
-
 Thermococcus kodakarensis
4.2.1.20 TrpB1 tryptophan synthase beta-subunit paralog Thermococcus kodakarensis
4.2.1.20 TrpB2 tryptophan synthase beta-subunit paralog Thermococcus kodakarensis
4.2.1.122 Trp synthase
-
 Thermococcus kodakarensis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.2.1.20 85
-
 assay at Thermococcus kodakarensis
4.2.1.122 85
-
 assay at Thermococcus kodakarensis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.2.1.20 0.39
-
 indole pH 7.5, 85°C, reaction catalyzed by TrpB2 Thermococcus kodakarensis
4.2.1.20 0.39
-
 indole pH 7.5, 85°C, reaction catalyzed by TrpB2 in presence of subunit TrpA Thermococcus kodakarensis
4.2.1.20 0.39
-
 L-serine subunit TrpB2 plus subunit TrpA, at pH 7.5 and 85°C Thermococcus kodakarensis
4.2.1.20 0.39
-
 L-serine subunit TrpB2, at pH 7.5 and 85°C Thermococcus kodakarensis
4.2.1.20 1.04
-
 indole pH 7.5, 85°C, reaction catalyzed by TrpB1 Thermococcus kodakarensis
4.2.1.20 1.04
-
 L-serine subunit TrpB1, at pH 7.5 and 85°C Thermococcus kodakarensis
4.2.1.20 3.46
-
 indole pH 7.5, 85°C, reaction catalyzed by TrpB1 in presence of subunit TrpA Thermococcus kodakarensis
4.2.1.20 3.46
-
 L-serine subunit TrpB1 plus subunit TrpA, at pH 7.5 and 85°C Thermococcus kodakarensis
4.2.1.122 0.39
-
 indole pH 7.5, 85°C, recombinant TrpB2, with or without TrpA Thermococcus kodakarensis
4.2.1.122 1.04
-
 indole pH 7.5, 85°C, recombinant TrpB1 Thermococcus kodakarensis
4.2.1.122 3.46
-
 indole pH 7.5, 85°C, recombinant TrpB1 with TrpA Thermococcus kodakarensis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
4.2.1.20 7.5
-
 assay at Thermococcus kodakarensis
4.2.1.122 7.5
-
 assay at Thermococcus kodakarensis

Cofactor

EC Number Cofactor Comment Organism Structure
4.2.1.122 pyridoxal 5'-phosphate
-
 Thermococcus kodakarensis

Expression

EC Number Organism Comment Expression
4.2.1.122 Thermococcus kodakarensis transcript level of trpB1 (within the trp operon) increases by 110fold in Trp(-) medium compared to those in Trp(+) medium up
4.2.1.122 Thermococcus kodakarensis transcript level of trpB2 increases by 45fold in Trp(-) medium compared to those in Trp(+) medium up

General Information

EC Number General Information Comment Organism
4.2.1.20 malfunction tryptophan synthase is composed of subunit TrpA and subunit TrpB. Thermococcus kodakarensis possess two paralogs of subunit B, TrpB1 and TrpB2. TrpB1 and TrpB2 both contribute to Trp biosynthesis. TrpB2 catalyzes the TrpB reaction but does not interact with TrpA as in the case of TrpB1. The double-deletion mutant (DELTAtrpB1DELTAtrpB2) displays Trp auxotrophy, whereas individual single mutants (DELTAB1 and DELTAB2 strains) do not Thermococcus kodakarensis
4.2.1.20 metabolism the last two steps of L-tryptophan biosynthesis are catalyzed by Trp synthase, a heterotetramer composed of TrpA and TrpB. TrpB catalyzes the condensation of indole, synthesized by TrpA, and serine to Trp. Thermococcus kodakarensis possesses two paralogs of subunit B, TrpB1 and TrpB2. TrpB1 and TrpB2 both contribute to Trp biosynthesis Thermococcus kodakarensis
4.2.1.122 malfunction the double-deletion mutant (DELTAtrpB1DELTAtrpB2) displays Trp auxotrophy, whereas individual single mutants (DELTAtrpB1 and DELTAtrpB2 strains) does not Thermococcus kodakarensis
4.2.1.122 metabolism the last two steps of L-tryptophan (Trp) biosynthesis are catalyzed by Trp synthase, a heterotetramer composed of TrpA and TrpB. TrpB catalyzes the condensation of indole, synthesized by TrpA, and serine to Trp. TrpB2 catalyzes the TrpB reaction but does not interact with TrpA as in the case of TrpB1. TrpB1 and TrpB2 both contribute to Trp biosynthesis in Thermococcus kodakarensis and can utilize free indole, and indole salvage does not necessarily rely on TrpB2 to a greater extent Thermococcus kodakarensis
4.2.1.122 physiological function trpB2 contributes in Trp biosynthesis Thermococcus kodakarensis

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
4.2.1.20 17
-
 indole pH 7.5, 85°C, reaction catalyzed by TrpB1 Thermococcus kodakarensis
4.2.1.20 17
-
 L-serine subunit TrpB2, at pH 7.5 and 85°C Thermococcus kodakarensis
4.2.1.20 48
-
 indole pH 7.5, 85°C, reaction catalyzed by TrpB2 Thermococcus kodakarensis
4.2.1.20 48
-
 L-serine subunit TrpB1, at pH 7.5 and 85°C Thermococcus kodakarensis
4.2.1.20 51
-
 indole pH 7.5, 85°C, reaction catalyzed by TrpB2 in presence of subunit TrpA Thermococcus kodakarensis
4.2.1.20 51
-
 L-serine subunit TrpB2 plus subunit TrpA, at pH 7.5 and 85°C Thermococcus kodakarensis
4.2.1.20 99
-
 indole pH 7.5, 85°C, reaction catalyzed by TrpB1 in presence of subunit TrpA Thermococcus kodakarensis
4.2.1.20 99
-
 L-serine subunit TrpB1 plus subunit TrpA, at pH 7.5 and 85°C Thermococcus kodakarensis
4.2.1.122 17
-
 indole pH 7.5, 85°C, recombinant TrpB1 Thermococcus kodakarensis
4.2.1.122 48
-
 indole pH 7.5, 85°C, recombinant TrpB2, without TrpA Thermococcus kodakarensis
4.2.1.122 51
-
 indole pH 7.5, 85°C, recombinant TrpB2, with TrpA Thermococcus kodakarensis
4.2.1.122 99
-
 indole pH 7.5, 85°C, recombinant TrpB1 with TrpA Thermococcus kodakarensis