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

BRENDA support

Literature summary extracted from

  • Francis, D.; Winn, M.; Latham, J.; Greaney, M.; Micklefield, J.
    An engineered tryptophan synthase opens new enzymatic pathways to beta-methyltryptophan and derivatives (2017), ChemBioChem, 18, 382-386 .
    View publication on PubMed

Application

EC Number Application Comment Organism
4.2.1.20 synthesis the reaction of mutant L166V with L-amino acid oxidase, halogenase enzymes and palladium chemocatalysts provides access to further D-configured and regioselectively halogenated or arylated beta-methyltryptophan derivatives Salmonella enterica subsp. enterica serovar Typhimurium

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.2.1.122 recombinant expression of C-terminally His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Salmonella enterica subsp. enterica serovar Typhimurium

Protein Variants

EC Number Protein Variants Comment Organism
4.2.1.20 L166A mutation in beta-subunit, 3fold increase in catalytic efficiency with L-threonine as substrate, decrease in efficiency with L-serine Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.20 L166V mutation in beta-subunit, 7fold increase in catalytic efficiency with L-threonine as substrate, decrease in efficiency with L-serine Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.122 L166A site-directed mutagenesis the beta-subunit, the mutant shows altered substrate specificity compared to the wild-type enzyme Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.122 L166V site-directed mutagenesis of the beta-subunit, the mutant shows altered substrate specificity compared to the wild-type enzyme, single-step enzymatic synthesis of beta-methyl-Trp derivatives, overview. Although 2-, 4-, 6- or 7-substituted indoles are accepted by StTrpS beta-L66V, along with L-Thr, 5-subsituted indoles prove to be very poor substrates for the enzyme. Variant beta-L166V can better accommodate L-Thr as a substrate Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.122 additional information an engineered variant of tryptophan synthase from Salmonella enterica (StTrpS) can catalyse the efficient condensation of L-threonine and various indoles to generate bmTrp and derivatives in a single step. Although L-serine is the natural substrate for TrpS, targeted mutagenesis of the StTrpS active site provides a variant (bL166V) that can better accommodate L-Thr as a substrate. The condensation of L-Thr and indole proceeds with retention of configuration at both alpha- and beta-positions to give (2S,3S)-beta-methyl-Trp Salmonella enterica subsp. enterica serovar Typhimurium

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.2.1.20 1.4
-
 L-serine mutant L166V, pH not specified in the publication, temperature not specified in the publication Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.20 1.6
-
 L-serine wild-type, pH not specified in the publication, temperature not specified in the publication Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.20 52
-
 L-serine mutant L166A, pH not specified in the publication, temperature not specified in the publication Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.20 210
-
 L-threonine wild-type, pH not specified in the publication, temperature not specified in the publication Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.20 310
-
 L-threonine mutant L166V, pH not specified in the publication, temperature not specified in the publication Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.20 340
-
 L-threonine mutant L166A, pH not specified in the publication, temperature not specified in the publication Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.122 1.4
-
 L-serine recombinant mutant L166V, pH 7.8, 37°C Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.122 1.6
-
 L-serine recombinant wild-type enzyme, pH 7.8, 37°C Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.122 52
-
 L-serine recombinant mutant L166A, pH 7.8, 37°C Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.122 210
-
 L-threonine recombinant wild-type enzyme, pH 7.8, 37°C Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.122 310
-
 L-threonine recombinant mutant L166V, pH 7.8, 37°C Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.122 340
-
 L-threonine recombinant mutant L166A, pH 7.8, 37°C Salmonella enterica subsp. enterica serovar Typhimurium

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4.2.1.122 L-serine + indole Salmonella enterica subsp. enterica serovar Typhimurium L-serine is the preferred substrate L-tryptophan + H2O
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
4.2.1.20 Salmonella enterica subsp. enterica serovar Typhimurium
-
-
-
4.2.1.122 Salmonella enterica subsp. enterica serovar Typhimurium
-
-
-
4.2.1.122 Salmonella enterica subsp. enterica serovar Typhimurium ATCC 37845
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.2.1.122 recombinant C-terminally His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21-A1(DE3) by nickel affinity chromatography, ultrafiltration, and desalting gel filtration Salmonella enterica subsp. enterica serovar Typhimurium

Reaction

EC Number Reaction Comment Organism Reaction ID
4.2.1.122 L-serine + indole = L-tryptophan + H2O reaction mechanism, overview Salmonella enterica subsp. enterica serovar Typhimurium
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.2.1.20 L-serine + indole
-
 Salmonella enterica subsp. enterica serovar Typhimurium L-tryptophan + H2O
-
 ?
4.2.1.20 L-serine + indole
-
 Salmonella enterica subsp. enterica serovar Typhimurium ATCC 37845 L-tryptophan + H2O
-
 ?
4.2.1.20 L-threonine + 2-methylindole
-
 Salmonella enterica subsp. enterica serovar Typhimurium 2-methyl-(2S,3S)-beta-methyltryptophan + H2O
-
 ?
4.2.1.20 L-threonine + 2-methylindole
-
 Salmonella enterica subsp. enterica serovar Typhimurium ATCC 37845 2-methyl-(2S,3S)-beta-methyltryptophan + H2O
-
 ?
4.2.1.20 L-threonine + 4-fluoroindole
-
 Salmonella enterica subsp. enterica serovar Typhimurium 4-fluoro-(2S,3S)-beta-methyltryptophan + H2O
-
 ?
4.2.1.20 L-threonine + 4-fluoroindole
-
 Salmonella enterica subsp. enterica serovar Typhimurium ATCC 37845 4-fluoro-(2S,3S)-beta-methyltryptophan + H2O
-
 ?
4.2.1.20 L-threonine + 4-methylindole
-
 Salmonella enterica subsp. enterica serovar Typhimurium 4-methyl-(2S,3S)-beta-methyltryptophan + H2O
-
 ?
4.2.1.20 L-threonine + 4-methylindole
-
 Salmonella enterica subsp. enterica serovar Typhimurium ATCC 37845 4-methyl-(2S,3S)-beta-methyltryptophan + H2O
-
 ?
4.2.1.20 L-threonine + 6-methylindole
-
 Salmonella enterica subsp. enterica serovar Typhimurium 6-methyl-(2S,3S)-beta-methyltryptophan + H2O
-
 ?
4.2.1.20 L-threonine + 7-chloroindole
-
 Salmonella enterica subsp. enterica serovar Typhimurium 7-chloro-(2S,3S)-beta-methyltryptophan + H2O
-
 ?
4.2.1.20 L-threonine + 7-fluoroindole
-
 Salmonella enterica subsp. enterica serovar Typhimurium 7-fluoro-(2S,3S)-beta-methyltryptophan + H2O
-
 ?
4.2.1.20 L-threonine + 7-methoxyindole
-
 Salmonella enterica subsp. enterica serovar Typhimurium 7-methoxy-(2S,3S)-beta-methyltryptophan + H2O
-
 ?
4.2.1.20 L-threonine + 7-methylindole
-
 Salmonella enterica subsp. enterica serovar Typhimurium 7-methyl-(2S,3S)-beta-methyltryptophan + H2O
-
 ?
4.2.1.20 L-threonine + indole
-
 Salmonella enterica subsp. enterica serovar Typhimurium (2S,3S)-beta-methyltryptophan + H2O
-
 ?
4.2.1.20 L-threonine + indole
-
 Salmonella enterica subsp. enterica serovar Typhimurium ATCC 37845 (2S,3S)-beta-methyltryptophan + H2O
-
 ?
4.2.1.122 L-serine + indole L-serine is the preferred substrate Salmonella enterica subsp. enterica serovar Typhimurium L-tryptophan + H2O
-
 ?
4.2.1.122 L-threonine + 2-methylindole
-
 Salmonella enterica subsp. enterica serovar Typhimurium (2S,3S)-2-methyl-beta-methyltryptophan + H2O
-
 ?
4.2.1.122 L-threonine + 4-fluoroindole
-
 Salmonella enterica subsp. enterica serovar Typhimurium (2S,3S)-4-fluoro-beta-methyltryptophan + H2O
-
 ?
4.2.1.122 L-threonine + 4-methylindole
-
 Salmonella enterica subsp. enterica serovar Typhimurium (2S,3S)-4-methyl-beta-methyltryptophan + H2O
-
 ?
4.2.1.122 L-threonine + 5-bromoindole
-
 Salmonella enterica subsp. enterica serovar Typhimurium (2S,3S)-5-bromo-beta-methyltryptophan + H2O
-
 ?
4.2.1.122 L-threonine + 5-chloroindole
-
 Salmonella enterica subsp. enterica serovar Typhimurium (2S,3S)-5-chloro-beta-methyltryptophan + H2O
-
 ?
4.2.1.122 L-threonine + 5-phenylindole
-
 Salmonella enterica subsp. enterica serovar Typhimurium (2S,3S)-5-phenyl-beta-methyltryptophan + H2O
-
 ?
4.2.1.122 L-threonine + 6-methylindole
-
 Salmonella enterica subsp. enterica serovar Typhimurium (2S,3S)-6-methyl-beta-methyltryptophan + H2O
-
 ?
4.2.1.122 L-threonine + 7-chloroindole
-
 Salmonella enterica subsp. enterica serovar Typhimurium (2S,3S)-7-chloro-beta-methyltryptophan + H2O
-
 ?
4.2.1.122 L-threonine + 7-fluoroindole
-
 Salmonella enterica subsp. enterica serovar Typhimurium (2S,3S)-7-fluoro-beta-methyltryptophan + H2O
-
 ?
4.2.1.122 L-threonine + 7-methoxyindole
-
 Salmonella enterica subsp. enterica serovar Typhimurium (2S,3S)-7-methoxy-beta-methyltryptophan + H2O
-
 ?
4.2.1.122 L-threonine + 7-methylindole
-
 Salmonella enterica subsp. enterica serovar Typhimurium (2S,3S)-7-methyl-beta-methyltryptophan + H2O
-
 ?
4.2.1.122 L-threonine + indole
-
 Salmonella enterica subsp. enterica serovar Typhimurium (2S,3S)-beta-methyltryptophan + H2O
-
 ?
4.2.1.122 additional information activity and isolated yields of L-beta-methyl-Trp are low compared to the wild-type reaction with l-Ser, which gives over 90% of L-Trp under the same conditions. Preperative scale synthesis (2S,3S)-L-beta-methyl tryptophan and derivatives Salmonella enterica subsp. enterica serovar Typhimurium ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
4.2.1.20 TRPS
-
 Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.122 StTrpS (beta2)
-
 Salmonella enterica subsp. enterica serovar Typhimurium

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.2.1.122 37
-
 assay at Salmonella enterica subsp. enterica serovar Typhimurium

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.2.1.20 0.0007
-
 L-threonine wild-type, pH not specified in the publication, temperature not specified in the publication Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.20 0.0035
-
 L-threonine mutant L166A, pH not specified in the publication, temperature not specified in the publication Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.20 0.007
-
 L-threonine mutant L166V, pH not specified in the publication, temperature not specified in the publication Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.20 0.0817
-
 L-serine mutant L166V, pH not specified in the publication, temperature not specified in the publication Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.20 0.183
-
 L-serine mutant L166A, pH not specified in the publication, temperature not specified in the publication Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.20 0.35
-
 L-serine wild-type, pH not specified in the publication, temperature not specified in the publication Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.122 0.00068
-
 L-threonine recombinant wild-type enzyme, pH 7.8, 37°C Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.122 0.0035
-
 L-threonine recombinant mutant L166A, pH 7.8, 37°C Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.122 0.007
-
 L-threonine recombinant mutant L166V, pH 7.8, 37°C Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.122 0.082
-
 L-serine recombinant mutant L166V, pH 7.8, 37°C Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.122 0.18
-
 L-serine recombinant mutant L166A, pH 7.8, 37°C Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.122 0.35
-
 L-serine recombinant wild-type enzyme, pH 7.8, 37°C Salmonella enterica subsp. enterica serovar Typhimurium

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
4.2.1.122 7.8
-
 assay at Salmonella enterica subsp. enterica serovar Typhimurium

General Information

EC Number General Information Comment Organism
4.2.1.122 additional information leucine residue Leu166 (betaL166) is in close proximity to the beta-position of the alpha-aminoacrylate species Salmonella enterica subsp. enterica serovar Typhimurium

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 0.0000032
-
 L-threonine wild-type, pH not specified in the publication, temperature not specified in the publication Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.20 0.0000102
-
 L-threonine mutant L166A, pH not specified in the publication, temperature not specified in the publication Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.20 0.000023
-
 L-threonine mutant L166V, pH not specified in the publication, temperature not specified in the publication Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.20 0.0037
-
 L-serine mutant L166A, pH not specified in the publication, temperature not specified in the publication Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.20 0.06
-
 L-serine mutant L166V, pH not specified in the publication, temperature not specified in the publication Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.20 0.217
-
 L-serine wild-type, pH not specified in the publication, temperature not specified in the publication Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.122 0.000003
-
 L-threonine recombinant wild-type enzyme, pH 7.8, 37°C Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.122 0.00001
-
 L-threonine recombinant mutant L166A, pH 7.8, 37°C Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.122 0.000023
-
 L-threonine recombinant mutant L166V, pH 7.8, 37°C Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.122 0.003
-
 L-serine recombinant mutant L166A, pH 7.8, 37°C Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.122 0.058
-
 L-serine recombinant mutant L166V, pH 7.8, 37°C Salmonella enterica subsp. enterica serovar Typhimurium
4.2.1.122 0.22
-
 L-serine recombinant wild-type enzyme, pH 7.8, 37°C Salmonella enterica subsp. enterica serovar Typhimurium