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Literature summary extracted from

  • Kotake, T.; Hojo, S.; Yamaguchi, D.; Aohara, T.; Konishi, T.; Tsumuraya, Y.
    Properties and physiological functions of UDP-sugar pyrophosphorylase in Arabidopsis (2007), Biosci. Biotechnol. Biochem., 71, 761-771.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.7.64 from cDNA generated from total RNA from 2-week-old seedlings in pGEM T-Easy for amplification and in pET32a for inducible expression with N-terminal thioredoxin- and His-tag in Escherichia coli BL21(DE3) and in pBI121 for generation of transgenic plants via Agrobacterium tumefaciens Arabidopsis thaliana

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.7.64 0.077
-
 UTP pH 7, 35°C Arabidopsis thaliana
2.7.7.64 0.094
-
 D-glucuronic acid 1-phosphate pH 7, 35°C Arabidopsis thaliana
2.7.7.64 0.16
-
 diphosphate pH 7, 35°C Arabidopsis thaliana
2.7.7.64 0.23
-
 D-glucose 1-phosphate pH 7, 35°C Arabidopsis thaliana
2.7.7.64 0.27
-
 D-galactose 1-phosphate pH 7, 35°C Arabidopsis thaliana
2.7.7.64 0.28
-
 UDP-glucose pH 7, 35°C Arabidopsis thaliana
2.7.7.64 1
-
 D-xylose 1-phosphate pH 7, 35°C Arabidopsis thaliana
2.7.7.64 1.5
-
 L-arabinose 1-phosphate pH 7, 35°C Arabidopsis thaliana

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.7.64 Mg2+ no activity in absence of divalent cations Arabidopsis thaliana
2.7.7.64 Mn2+ no activity in absence of divalent cations Arabidopsis thaliana

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.7.7.64 67850
-
 deduced from amino acid sequence Arabidopsis thaliana
2.7.7.64 70000
-
 purified recombinant enzyme, SDS-PAGE Arabidopsis thaliana

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.7.64 UDP-monosaccharide + diphosphate Arabidopsis thaliana salvage pathway monosaccharide 1-phosphate + UTP
-
 r

Organism

EC Number Organism UniProt Comment Textmining
2.7.7.64 Arabidopsis thaliana Q9C5I1 ecotype Columbia, Landsberg erecta, Wassilewskija
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.7.64 chelating chromatography (elution: 250 mM imidazole) followed by cleavage of N-terminal thioredoxin- and His-tag with thrombin and DEAE Sepharose FF chromatography Arabidopsis thaliana

Source Tissue

EC Number Source Tissue Comment Organism Textmining
2.7.7.64 aerial part enzyme assay on homogenate of transgenic plants Arabidopsis thaliana
-
2.7.7.64 cauline leaf AtUSP mRNA-specific qRT-PCR Arabidopsis thaliana
-
2.7.7.64 cotyledon AtUSP mRNA-specific qRT-PCR, strong expression in vascular tissue (promoter: beta-glucuronidase assay) Arabidopsis thaliana
-
2.7.7.64 flower relatively high levels in most cell types (AtUSP mRNA-specific qRT-PCR) Arabidopsis thaliana
-
2.7.7.64 inflorescence stem AtUSP mRNA-specific qRT-PCR Arabidopsis thaliana
-
2.7.7.64 leaf young leaves (AtUSP mRNA-specific qRT-PCR), strong expression in vascular tissue (promoter: beta-glucuronidase assay) Arabidopsis thaliana
-
2.7.7.64 pollen AtUSP mRNA-specific qRT-PCR, strong expression in vascular tissue (promoter: beta-glucuronidase assay), essential for pollen development since loss-of-function mutation in enzyme causes male infertility Arabidopsis thaliana
-
2.7.7.64 root strong expression in growing region (promoter: beta-glucuronidase assay), young roots (AtUSP mRNA-specific qRT-PCR) Arabidopsis thaliana
-
2.7.7.64 rosette leaf AtUSP mRNA-specific qRT-PCR Arabidopsis thaliana
-
2.7.7.64 seedling young seedlings, expression in most cell types (AtUSP mRNA-specific qRT-PCR) Arabidopsis thaliana
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.7.7.64 additional information
-
 no activity detectable for substrates L-fucose 1-phosphate and mannose 1-phosphate Arabidopsis thaliana
2.7.7.64 additional information
-
 143-255% UDP-L-arabinose synthesising activity in extracts from transgenic plants (aerial parts) compared to wild-type plant Arabidopsis thaliana
2.7.7.64 additional information
-
 25% of wild-type UDP-L-arabinose synthesising activity in plant extracts (aerial parts) upon repression of AtUSP gene expression Arabidopsis thaliana
2.7.7.64 1.2
-
 +/-0.08 micromol/min/mg, N-acetylglucosamine 1-phosphate Arabidopsis thaliana
2.7.7.64 22.8
-
 +/-1.1 micromol/min/mg, D-xylose 1-phosphate Arabidopsis thaliana
2.7.7.64 44.5
-
 +/-0.9 micromol/min/mg, L-arabinose 1-phosphate Arabidopsis thaliana
2.7.7.64 51.4
-
 +/-3.6 micromol/min/mg, D-glucuronic acid 1-phosphate Arabidopsis thaliana
2.7.7.64 84.3
-
 +/-2.4 micromol/min/mg, D-glucose 1-phosphate Arabidopsis thaliana
2.7.7.64 86.9
-
 +/-2.4 micromol/min/mg, D-galactose 1-phosphate Arabidopsis thaliana

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.7.64 D-galactose 1-phosphate + UTP
-
 Arabidopsis thaliana UDP-galactose + diphosphate
-
 ?
2.7.7.64 D-glucose 1-phosphate + UTP pH 7, 35°C, 2 mM Mg2+ Arabidopsis thaliana UDP-glucose + diphosphate reaction stop by boiling, analysis of products by HPLC (monitoring at 262 nm) r
2.7.7.64 D-glucuronic acid 1-phosphate + UTP
-
 Arabidopsis thaliana UDP-glucuronic acid + diphosphate
-
 ?
2.7.7.64 D-xylose 1-phosphate + UTP
-
 Arabidopsis thaliana UDP-xylose + diphosphate
-
 ?
2.7.7.64 L-arabinose 1-phosphate + UTP
-
 Arabidopsis thaliana UDP-L-arabinose + diphosphate
-
 ?
2.7.7.64 additional information no activity detectable for substrates L-fucose 1-phosphate and mannose 1-phosphate Arabidopsis thaliana ?
-
 ?
2.7.7.64 N-acetylglucosamine 1-phosphate + UTP poorly catalysed reaction Arabidopsis thaliana UDP-N-acetylglucosamine + diphosphate
-
 ?
2.7.7.64 UDP-monosaccharide + diphosphate salvage pathway Arabidopsis thaliana monosaccharide 1-phosphate + UTP
-
 r

Synonyms

EC Number Synonyms Comment Organism
2.7.7.64 AtUSP
-
 Arabidopsis thaliana
2.7.7.64 UDP-sugar pyrophosphorylase
-
 Arabidopsis thaliana
2.7.7.64 USP
-
 Arabidopsis thaliana

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.7.64 45
-
 UDP-glucose-synthesising activity Arabidopsis thaliana

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.7.7.64 55
-
 loss of UDP-glucose-synthesising activity Arabidopsis thaliana

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.7.64 additional information
-
 D-glucose 1-phosphate Kcat/KM: 530000 1/M/sec, pH 7, 35°C Arabidopsis thaliana
2.7.7.64 additional information
-
 D-galactose 1-phosphate Kcat/KM: 490000 1/M/sec, pH 7, 35°C Arabidopsis thaliana
2.7.7.64 additional information
-
 D-glucuronic acid 1-phosphate Kcat/KM: 710000 1/M/sec, pH 7, 35°C Arabidopsis thaliana
2.7.7.64 additional information
-
 L-arabinose 1-phosphate Kcat/KM: 92000 1/M/sec, pH 7, 35°C Arabidopsis thaliana
2.7.7.64 additional information
-
 D-xylose 1-phosphate Kcat/KM: 51000 1/M/sec, pH 7, 35°C Arabidopsis thaliana
2.7.7.64 additional information
-
 UTP Kcat/KM: 1400000 1/M/sec, pH 7, 35°C Arabidopsis thaliana
2.7.7.64 additional information
-
 UDP-glucose Kcat/KM: 480000 1/M/sec, pH 7, 35°C Arabidopsis thaliana
2.7.7.64 additional information
-
 diphosphate Kcat/KM: 860000 1/M/sec, pH 7, 35°C Arabidopsis thaliana
2.7.7.64 51
-
 D-xylose 1-phosphate pH 7, 35°C Arabidopsis thaliana
2.7.7.64 67
-
 D-glucuronic acid 1-phosphate pH 7, 35°C Arabidopsis thaliana
2.7.7.64 108
-
 UTP pH 7, 35°C Arabidopsis thaliana
2.7.7.64 122
-
 D-glucose 1-phosphate pH 7, 35°C Arabidopsis thaliana
2.7.7.64 133
-
 D-galactose 1-phosphate pH 7, 35°C Arabidopsis thaliana
2.7.7.64 133
-
 UDP-glucose pH 7, 35°C Arabidopsis thaliana
2.7.7.64 138
-
 L-arabinose 1-phosphate pH 7, 35°C Arabidopsis thaliana
2.7.7.64 138
-
 diphosphate pH 7, 35°C Arabidopsis thaliana

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.7.64 7
-
 UDP-glucose-synthesising activity Arabidopsis thaliana

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.7.7.64 6 8 60% of maximum UDP-glucose-synthesising activity Arabidopsis thaliana

pI Value

EC Number Organism Comment pI Value Maximum pI Value
2.7.7.64 Arabidopsis thaliana theoretical
-
 6.1