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Literature summary extracted from

  • Kumari, K.; Tlapak-Simmons, V.L.; Baggenstoss, B.A.; Weigel, P.H.
    The streptococcal hyaluronan synthases are inhibited by sulfhydryl-modifying reagents, but conserved cysteine residues are not essential for enzyme function (2002), J. Biol. Chem., 277, 13943-13951.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.4.1.212 expression of wild-type and mutant enzymes as His-tagged proteins in Escherichia coli Streptococcus dysgalactiae subsp. equisimilis

Protein Variants

EC Number Protein Variants Comment Organism
2.4.1.212 C226A site-directed mutagenesis, 24% remaining activity and altered kinetic constants compared to the wild-type enzyme Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 C226A/C262A site-directed mutagenesis, 3.2% remaining activity and altered kinetic constants compared to the wild-type enzyme Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 C226A/C262A/C367A site-directed mutagenesis, 1.4% remaining activity and altered kinetic constants compared to the wild-type enzyme Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 C226A/C281A site-directed mutagenesis, reduced reaction velocity and altered Km values compared to the wild-type enzyme Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 C226A/C367A site-directed mutagenesis, reduced reaction velocity and altered Km values compared to the wild-type enzyme Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 C226S site-directed mutagenesis, highly reduced reaction velocity and altered Km values compared to the wild-type enzyme Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 C262A site-directed mutagenesis, reduced reaction velocity and increased Km values compared to the wild-type enzyme Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 C262A/C281A site-directed mutagenesis, reduced reaction velocity and altered Km values compared to the wild-type enzyme Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 C262A/C367A site-directed mutagenesis, reduced reaction velocity and altered Km values compared to the wild-type enzyme Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 C262S site-directed mutagenesis, reduced reaction velocity and increased Km values compared to the wild-type enzyme Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 C281A site-directed mutagenesis, reduced reaction velocity and altered Km values compared to the wild-type enzyme Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 C281A/C367A site-directed mutagenesis, reduced reaction velocity and altered Km values compared to the wild-type enzyme Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 C281S site-directed mutagenesis, reduced reaction velocity and altered Km values compared to the wild-type enzyme Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 C367A site-directed mutagenesis, increased reaction velocity and Km values compared to the wild-type enzyme Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 C367S site-directed mutagenesis, kinetics similar to the wild-type enzyme Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 additional information construction and kinetic analysis of Cys-deletion mutants, overview Streptococcus dysgalactiae subsp. equisimilis

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.4.1.212 DTNB 52% inhibition at 0.5 mM Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 DTNB 52% inhibition at 0.5 mM Streptococcus pyogenes
2.4.1.212 iodoacetamide 15% inhibition at 5 mM, biphasic inhibition Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 iodoacetamide 13% inhibition at 5 mM, biphasic inhibition Streptococcus pyogenes
2.4.1.212 methylmethanethiosulfonate 93% reduced activity at 0.05 mM Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 methylmethanethiosulfonate 89% reduced activity at 0.05 mM Streptococcus pyogenes
2.4.1.212 N-ethylmaleimide biphasic inhibition, about 50% reduced reaction velocity, unaltered Km-value, 70% inhibition at 5 mM Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 N-ethylmaleimide 60% inhibition at 5 mM, biphasic inhibition Streptococcus pyogenes
2.4.1.212 sodium arsenite 40% inhibition at 10 mM Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 sodium arsenite 46% inhibition at 10 mM Streptococcus pyogenes
2.4.1.212 sulfhydryl reagents enzyme is inhibited by sulfhydryl-modifying reagents, inhibition mode and mechanism Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 sulfhydryl reagents enzyme is inhibited by sulfhydryl-modifying reagents, inhibition mode and mechanism Streptococcus pyogenes

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.4.1.212 additional information
-
 additional information kinetics, kinetic analysis of Cys-deletion mutants, overview Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 0.04
-
 UDP-D-glucuronate pH 7.0, 30°C, recombinant mutant C281A Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 0.044
-
 UDP-D-glucuronate pH 7.0, 30°C, recombinant mutant C226S Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 0.053
-
 UDP-N-acetyl-D-glucosamine pH 7.0, 30°C, recombinant mutant C226A/C281A Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 0.056
-
 UDP-D-glucuronate pH 7.0, 30°C, recombinant mutant C281S Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 0.065
-
 UDP-N-acetyl-D-glucosamine pH 7.0, 30°C, recombinant mutant C281A/C367A Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 0.074
-
 UDP-N-acetyl-D-glucosamine pH 7.0, 30°C, recombinant wild-type enzyme Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 0.077
-
 UDP-D-glucuronate pH 7.0, 30°C, recombinant wild-type enzyme Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 0.079
-
 UDP-N-acetyl-D-glucosamine pH 7.0, 30°C, recombinant mutant C226A/C367A Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 0.079
-
 UDP-D-glucuronate pH 7.0, 30°C, recombinant mutant C367S Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 0.085
-
 UDP-D-glucuronate pH 7.0, 30°C, recombinant mutant C367A Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 0.088
-
 UDP-D-glucuronate pH 7.0, 30°C, recombinant mutant C226A Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 0.09
-
 UDP-N-acetyl-D-glucosamine pH 7.0, 30°C, recombinant mutant C367A Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 0.091
-
 UDP-N-acetyl-D-glucosamine pH 7.0, 30°C, recombinant mutant C367S Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 0.096
-
 UDP-D-glucuronate pH 7.0, 30°C, recombinant mutant C262S Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 0.098
-
 UDP-N-acetyl-D-glucosamine pH 7.0, 30°C, recombinant mutant C281S Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 0.113
-
 UDP-N-acetyl-D-glucosamine pH 7.0, 30°C, recombinant mutant C262A/C281A Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 0.121
-
 UDP-N-acetyl-D-glucosamine pH 7.0, 30°C, recombinant mutant C262A/C367A Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 0.13
-
 UDP-N-acetyl-D-glucosamine pH 7.0, 30°C, recombinant mutant C281A Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 0.134
-
 UDP-N-acetyl-D-glucosamine pH 7.0, 30°C, recombinant mutant C226A/C262A Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 0.146
-
 UDP-D-glucuronate pH 7.0, 30°C, recombinant mutant C262A Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 0.153
-
 UDP-N-acetyl-D-glucosamine pH 7.0, 30°C, recombinant mutant C262S Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 0.154
-
 UDP-N-acetyl-D-glucosamine pH 7.0, 30°C, recombinant mutant C226A Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 0.186
-
 UDP-N-acetyl-D-glucosamine pH 7.0, 30°C, recombinant mutant C262A Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 0.232
-
 UDP-N-acetyl-D-glucosamine pH 7.0, 30°C, recombinant mutant C226S Streptococcus dysgalactiae subsp. equisimilis

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
2.4.1.212 membrane bound Streptococcus dysgalactiae subsp. equisimilis 16020
-
2.4.1.212 membrane bound Streptococcus pyogenes 16020
-

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.4.1.212 UDP-N-acetyl-D-glucosamine + UDP-D-glucuronate Streptococcus dysgalactiae subsp. equisimilis biosynthesis of hyaluronan [beta-N-acetyl-D-glucosaminyl(1-4)beta-D-glucuronosyl(1-3)]n + UDP
-
 ?
2.4.1.212 UDP-N-acetyl-D-glucosamine + UDP-D-glucuronate Streptococcus pyogenes biosynthesis of hyaluronan [beta-N-acetyl-D-glucosaminyl(1-4)beta-D-glucuronosyl(1-3)]n + UDP
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.4.1.212 Streptococcus dysgalactiae subsp. equisimilis
-
-
-
2.4.1.212 Streptococcus pyogenes Q5X9A9
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.4.1.212 recombinant His-tagged wild-type and mutant enzymes from Escherichia coli Streptococcus dysgalactiae subsp. equisimilis

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.4.1.212 additional information
-
-
 Streptococcus dysgalactiae subsp. equisimilis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.1.212 additional information conserved cysteine residues are not essential for enzyme function Streptococcus dysgalactiae subsp. equisimilis ?
-
 ?
2.4.1.212 additional information conserved cysteine residues are not essential for enzyme function Streptococcus pyogenes ?
-
 ?
2.4.1.212 UDP-N-acetyl-D-glucosamine + UDP-D-glucuronate
-
 Streptococcus dysgalactiae subsp. equisimilis [beta-N-acetyl-D-glucosaminyl(1-4)beta-D-glucuronosyl(1-3)]n + UDP
-
 ?
2.4.1.212 UDP-N-acetyl-D-glucosamine + UDP-D-glucuronate
-
 Streptococcus pyogenes [beta-N-acetyl-D-glucosaminyl(1-4)beta-D-glucuronosyl(1-3)]n + UDP
-
 ?
2.4.1.212 UDP-N-acetyl-D-glucosamine + UDP-D-glucuronate biosynthesis of hyaluronan Streptococcus dysgalactiae subsp. equisimilis [beta-N-acetyl-D-glucosaminyl(1-4)beta-D-glucuronosyl(1-3)]n + UDP
-
 ?
2.4.1.212 UDP-N-acetyl-D-glucosamine + UDP-D-glucuronate biosynthesis of hyaluronan Streptococcus pyogenes [beta-N-acetyl-D-glucosaminyl(1-4)beta-D-glucuronosyl(1-3)]n + UDP
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.4.1.212 HAS
-
 Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 HAS
-
 Streptococcus pyogenes
2.4.1.212 More the enzyme belongs to the group of class I hyaluronan synthases Streptococcus dysgalactiae subsp. equisimilis
2.4.1.212 More the enzyme belongs to the group of class I hyaluronan synthases Streptococcus pyogenes

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.4.1.212 30 37 assay at Streptococcus dysgalactiae subsp. equisimilis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.4.1.212 7
-
 assay at Streptococcus dysgalactiae subsp. equisimilis