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Literature summary extracted from

  • Nakamichi, Y.; Mikami, B.; Murata, K.; Hashimoto, W.
    Crystal structure of a bacterial unsaturated glucuronyl hydrolase with specificity for heparin (2014), J. Biol. Chem., 289, 4787-4797.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.180 gene Phep_2238, DNA and amino acid sequence determination and analysis, sequence comparisons, expression in Escherichia coli strain BL21(DE3) Pedobacter heparinus
3.2.1.180 gene Phep_2649, DNA and amino acid sequence determination and analysis, sequence comparisons, expression in Escherichia coli strain BL21(DE3) Pedobacter heparinus
3.2.1.180 gene Phep_2830, DNA and amino acid sequence determination and analysis, sequence comparisons, expression of tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Pedobacter heparinus
3.2.1.180 sequence comparisons Bacillus sp. (in: Bacteria)
3.2.1.180 sequence comparisons, recombinant expression in Escherichia coli Streptococcus pneumoniae
3.2.1.180 sequence comparisons, recombinant expression in Escherichia coli Streptococcus pyogenes
3.2.1.180 sequence comparisons, recombinant expression of wild-type and mutant enzymes in Escherichia coli Streptococcus agalactiae

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.2.1.180 purified recombinant wild-type enzyme, sitting drop vapor diffusion method, mixing of 0..001 ml of 10 mg/ml protein in n 20 mM Tris-HCl, pH 7.5, 1 mM dithiothreitol, and 1 mM EDTA, with 0.001 ml of reservoir solution containing 25% PEG 6000, 0.1 M HEPES, pH 7.5, and 0.1 M lithium chloride, 20°C, 2 months, X-ray diffraction structure determination and analysis at 1.3 A resolution, molecular replacement Pedobacter heparinus

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.180 D174N site-directed mutagenesis Pedobacter heparinus
3.2.1.180 D182N site-directed mutagenesis Pedobacter heparinus
3.2.1.180 F164A site-directed mutagenesis Pedobacter heparinus
3.2.1.180 G362Y site-directed mutagenesis Pedobacter heparinus
3.2.1.180 K370A site-directed mutagenesis Streptococcus agalactiae
3.2.1.180 R57A site-directed mutagenesis Pedobacter heparinus
3.2.1.180 S365A site-directed mutagenesis Streptococcus agalactiae
3.2.1.180 S365G site-directed mutagenesis Streptococcus agalactiae
3.2.1.180 S368A site-directed mutagenesis Streptococcus agalactiae
3.2.1.180 S368G site-directed mutagenesis Streptococcus agalactiae

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.180 0.029
-
 DELTAGlcUA-beta-(1->4)-GlcNAc pH 7.5, 30°C, recombinant wild-type enzyme Pedobacter heparinus
3.2.1.180 0.86
-
 DELTAGlcUA-beta-(1->4)-GlcN2S pH 7.5, 30°C, recombinant mutant S370A Streptococcus agalactiae
3.2.1.180 1.4
-
 DELTAGlcUA-beta-(1->4)-GlcNAc pH 7.5, 30°C, recombinant mutant F164A Pedobacter heparinus
3.2.1.180 1.9
-
 DELTAGlcUA-beta-(1->4)-GlcN2S pH 7.5, 30°C, recombinant wild-type enzyme Streptococcus agalactiae
3.2.1.180 3
-
 DELTAGlcUA-beta-(1->4)-GlcN2S pH 7.5, 30°C, recombinant mutant S368A Streptococcus agalactiae
3.2.1.180 3.4
-
 DELTAGlcUA-beta-(1->4)-GlcN2S pH 7.5, 30°C, recombinant mutant S365A Streptococcus agalactiae
3.2.1.180 3.4
-
 DELTAGlcUA-beta-(1->4)-GlcN2S pH 7.5, 30°C, recombinant mutant S368G Streptococcus agalactiae
3.2.1.180 5.2
-
 DELTAGlcUA-beta-(1->4)-GlcN2S pH 7.5, 30°C, recombinant mutant S365G Streptococcus agalactiae

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.180 Bacillus sp. (in: Bacteria)
-
-
-
3.2.1.180 Pedobacter heparinus C6XY24 gene Phep_2238
-
3.2.1.180 Pedobacter heparinus C6Y0D8 gene Phep_2649
-
3.2.1.180 Pedobacter heparinus C6Y1N7 gene Phep_2830
-
3.2.1.180 Streptococcus agalactiae
-
-
-
3.2.1.180 Streptococcus pneumoniae
-
-
-
3.2.1.180 Streptococcus pyogenes
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.180 recombinant enzyme from Escherichia coli Streptococcus pneumoniae
3.2.1.180 recombinant enzyme from Escherichia coli Streptococcus pyogenes
3.2.1.180 recombinant tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by affinity chromatography to near homogeneity Pedobacter heparinus
3.2.1.180 recombinant wild-type and mutant enzymes from Escherichia coli Streptococcus agalactiae

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.2.1.180 1.7
-
 purified recombinant wild-type enzyme, pH 7.5, 30°C, substrate DELTAGlcUA-beta-(1->3)-GalNAc6S Pedobacter heparinus
3.2.1.180 5.3
-
 purified recombinant wild-type enzyme, pH 7.5, 30°C, substrate DELTAGlcUA-beta-(1->3)-GalNAc with a sulfate group at the C-6 position of GalNAc Streptococcus pyogenes
3.2.1.180 5.3
-
 purified recombinant wild-type enzyme, pH 7.5, 30°C, substrate DELTAGlcUA-beta-(1->4)-GlcNAc Bacillus sp. (in: Bacteria)
3.2.1.180 6.1
-
 purified recombinant wild-type enzyme, pH 7.5, 30°C, substrate DELTAGlcUA-beta-(1,3)-GalNAc6S Streptococcus pneumoniae
3.2.1.180 12
-
 purified recombinant wild-type enzyme, pH 7.5, 30°C, substrate DELTAGlcUA-beta-(1->4)-GlcNAc Pedobacter heparinus
3.2.1.180 15
-
 purified recombinant wild-type enzyme, pH 7.5, 30°C, substrate DELTAGlcUA-beta-(1->4)-GlcNAc Pedobacter heparinus
3.2.1.180 17
-
 purified recombinant wild-type enzyme, pH 7.5, 30°C, substrate DELTAGlcUA-beta-(1->3)-GalNAc6S Streptococcus agalactiae

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.180 DELTA4GlcUA2S-beta-(1->4)-GlcN2S + H2O
-
 Streptococcus agalactiae ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1,3)-GalNAc + H2O
-
 Streptococcus pneumoniae ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1,3)-GalNAc4S + H2O
-
 Streptococcus pneumoniae ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1,3)-GalNAc6S + H2O best substrate Streptococcus pneumoniae ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1,4)-GlcN2S + H2O
-
 Streptococcus pneumoniae ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1,4)-GlcN2S6S + H2O
-
 Streptococcus pneumoniae ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1,4)-GlcN6S + H2O
-
 Streptococcus pneumoniae ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1,4)-GlcNAc + H2O
-
 Streptococcus pneumoniae ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1,4)-GlcNAc6S + H2O
-
 Streptococcus pneumoniae ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1->3)-GalNAc + H2O
-
 Streptococcus pyogenes ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1->3)-GalNAc + H2O
-
 Bacillus sp. (in: Bacteria) ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1->3)-GalNAc + H2O
-
 Streptococcus agalactiae ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1->3)-GalNAc + H2O
-
 Pedobacter heparinus ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1->3)-GalNAc4S + H2O
-
 Streptococcus pyogenes ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1->3)-GalNAc4S + H2O
-
 Streptococcus agalactiae ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1->3)-GalNAc4S + H2O
-
 Pedobacter heparinus ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1->3)-GalNAc6S + H2O
-
 Bacillus sp. (in: Bacteria) ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1->3)-GalNAc6S + H2O
-
 Pedobacter heparinus ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1->3)-GalNAc6S + H2O best substrate Streptococcus pyogenes ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1->3)-GalNAc6S + H2O best substrate Streptococcus agalactiae ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1->3)-GalNAc6S + H2O best substrate Pedobacter heparinus ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1->4)-GlcN2S + H2O
-
 Streptococcus pyogenes ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1->4)-GlcN2S + H2O
-
 Bacillus sp. (in: Bacteria) ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1->4)-GlcN2S + H2O
-
 Streptococcus agalactiae ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1->4)-GlcN2S + H2O
-
 Pedobacter heparinus ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1->4)-GlcN2S6S + H2O
-
 Bacillus sp. (in: Bacteria) ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1->4)-GlcN2S6S + H2O
-
 Streptococcus agalactiae ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1->4)-GlcN2S6S + H2O
-
 Pedobacter heparinus ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1->4)-GlcN6S + H2O
-
 Streptococcus pyogenes ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1->4)-GlcN6S + H2O
-
 Bacillus sp. (in: Bacteria) ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1->4)-GlcN6S + H2O
-
 Streptococcus agalactiae ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1->4)-GlcN6S + H2O
-
 Pedobacter heparinus ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1->4)-GlcNAc + H2O
-
 Streptococcus pyogenes ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1->4)-GlcNAc + H2O
-
 Streptococcus agalactiae ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1->4)-GlcNAc + H2O
-
 Pedobacter heparinus ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1->4)-GlcNAc + H2O best substrate Bacillus sp. (in: Bacteria) ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1->4)-GlcNAc + H2O best substrate Pedobacter heparinus ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1->4)-GlcNAc6S + H2O
-
 Streptococcus pyogenes ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1->4)-GlcNAc6S + H2O
-
 Bacillus sp. (in: Bacteria) ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1->4)-GlcNAc6S + H2O
-
 Streptococcus agalactiae ?
-
 ?
3.2.1.180 DELTAGlcUA-beta-(1->4)-GlcNAc6S + H2O
-
 Pedobacter heparinus ?
-
 ?
3.2.1.180 DELTAGlcUA2S-beta-(1,4)-GlcN2S + H2O
-
 Streptococcus pneumoniae ?
-
 ?
3.2.1.180 DELTAGlcUA2S-beta-(1->4)-GlcN2S + H2O
-
 Bacillus sp. (in: Bacteria) ?
-
 ?
3.2.1.180 additional information substrate specificity of type and mutant enzymes with unsaturated chondroitin disaccharides and unsaturated heparin disaccharides, substrate docking studies, overview Pedobacter heparinus ?
-
 ?
3.2.1.180 additional information substrate specificity of wild-type and mutant enzymes with unsaturated chondroitin disaccharides and unsaturated heparin disaccharides, overview Streptococcus agalactiae ?
-
 ?
3.2.1.180 additional information substrate specificity with unsaturated chondroitin disaccharides and unsaturated heparin disaccharides, overview Streptococcus pneumoniae ?
-
 ?
3.2.1.180 additional information substrate specificity with unsaturated chondroitin disaccharides and unsaturated heparin disaccharides, overview Streptococcus pyogenes ?
-
 ?
3.2.1.180 additional information substrate specificity with unsaturated chondroitin disaccharides and unsaturated heparin disaccharides, overview Bacillus sp. (in: Bacteria) ?
-
 ?
3.2.1.180 additional information substrate specificity with unsaturated chondroitin disaccharides and unsaturated heparin disaccharides, overview Pedobacter heparinus ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.180 More three-dimensional structure analysis, overview Pedobacter heparinus

Synonyms

EC Number Synonyms Comment Organism
3.2.1.180 Phep_2238
-
 Pedobacter heparinus
3.2.1.180 Phep_2649
-
 Pedobacter heparinus
3.2.1.180 Phep_2830
-
 Pedobacter heparinus
3.2.1.180 SagUGL
-
 Streptococcus agalactiae
3.2.1.180 SpnUGL
-
 Streptococcus pneumoniae
3.2.1.180 SpyUGL
-
 Streptococcus pyogenes
3.2.1.180 UGL
-
 Streptococcus pneumoniae
3.2.1.180 UGL
-
 Streptococcus pyogenes
3.2.1.180 UGL
-
 Bacillus sp. (in: Bacteria)
3.2.1.180 UGL
-
 Streptococcus agalactiae
3.2.1.180 UGL
-
 Pedobacter heparinus
3.2.1.180 unsaturated glucuronyl hydrolase
-
 Streptococcus pneumoniae
3.2.1.180 unsaturated glucuronyl hydrolase
-
 Streptococcus pyogenes
3.2.1.180 unsaturated glucuronyl hydrolase
-
 Bacillus sp. (in: Bacteria)
3.2.1.180 unsaturated glucuronyl hydrolase
-
 Streptococcus agalactiae
3.2.1.180 unsaturated glucuronyl hydrolase
-
 Pedobacter heparinus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.180 30
-
 assay at Streptococcus pneumoniae
3.2.1.180 30
-
 assay at Streptococcus pyogenes
3.2.1.180 30
-
 assay at Bacillus sp. (in: Bacteria)
3.2.1.180 30
-
 assay at Streptococcus agalactiae
3.2.1.180 30
-
 assay at Pedobacter heparinus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.180 0.075
-
 DELTAGlcUA-beta-(1->4)-GlcN2S pH 7.5, 30°C, recombinant mutant S370A Streptococcus agalactiae
3.2.1.180 0.76
-
 DELTAGlcUA-beta-(1->4)-GlcN2S pH 7.5, 30°C, recombinant mutant S365G Streptococcus agalactiae
3.2.1.180 1.9
-
 DELTAGlcUA-beta-(1->4)-GlcN2S pH 7.5, 30°C, recombinant mutant S365A Streptococcus agalactiae
3.2.1.180 2.8
-
 DELTAGlcUA-beta-(1->4)-GlcN2S pH 7.5, 30°C, recombinant mutant S368G Streptococcus agalactiae
3.2.1.180 3.8
-
 DELTAGlcUA-beta-(1->4)-GlcN2S pH 7.5, 30°C, recombinant wild-type enzyme Streptococcus agalactiae
3.2.1.180 6
-
 DELTAGlcUA-beta-(1->4)-GlcN2S pH 7.5, 30°C, recombinant mutant S368A Streptococcus agalactiae
3.2.1.180 16
-
 DELTAGlcUA-beta-(1->4)-GlcNAc pH 7.5, 30°C, recombinant wild-type enzyme Pedobacter heparinus
3.2.1.180 22
-
 DELTAGlcUA-beta-(1->4)-GlcNAc pH 7.5, 30°C, recombinant mutant F164A Pedobacter heparinus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.180 7.5
-
 assay at Streptococcus pneumoniae
3.2.1.180 7.5
-
 assay at Streptococcus pyogenes
3.2.1.180 7.5
-
 assay at Bacillus sp. (in: Bacteria)
3.2.1.180 7.5
-
 assay at Streptococcus agalactiae
3.2.1.180 7.5
-
 assay at Pedobacter heparinus

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.2.1.180 0.086
-
 DELTAGlcUA-beta-(1->4)-GlcN2S pH 7.5, 30°C, recombinant mutant S370A Streptococcus agalactiae
3.2.1.180 0.15
-
 DELTAGlcUA-beta-(1->4)-GlcN2S pH 7.5, 30°C, recombinant mutant S365G Streptococcus agalactiae
3.2.1.180 0.56
-
 DELTAGlcUA-beta-(1->4)-GlcN2S pH 7.5, 30°C, recombinant mutant S365A Streptococcus agalactiae
3.2.1.180 0.83
-
 DELTAGlcUA-beta-(1->4)-GlcN2S pH 7.5, 30°C, recombinant mutant S368G Streptococcus agalactiae
3.2.1.180 2
-
 DELTAGlcUA-beta-(1->4)-GlcN2S pH 7.5, 30°C, recombinant wild-type enzyme Streptococcus agalactiae
3.2.1.180 2
-
 DELTAGlcUA-beta-(1->4)-GlcN2S pH 7.5, 30°C, recombinant mutant S368A Streptococcus agalactiae