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Literature summary for 2.4.1.128 extracted from

  • Weis, M.; Lim, E.K.; Bruce, N.C.; Bowles, D.J.
    Engineering and kinetic characterisation of two glucosyltransferases from Arabidopsis thaliana (2008), Biochimie, 90, 830-834.
    View publication on PubMed
Search for more literature for 2.4.1.128

Cloned(Commentary)

Cloned (Comment) Organism
recombinant and mutant glycosyltransferase 71C1 is expressed as fusion protein tagged with a 26 kDa GST protein at the N-terminus in Escherichia coli BL21, production of a chimeric mutant derived from 2 Arabidopsis glucosyltransferases, 71C1 and 71C3, the chimera contains the N-terminal domain of 71C1 and the C-terminal domain of 71C3 Arabidopsis thaliana
recombinant and mutant glycosyltransferase 71C3 is expressed as fusion protein tagged with a 26 kDa GST protein at the N-terminus in Escherichia coli BL21, production of a chimeric mutant derived from 2 Arabidopsis glucosyltransferases, 71C1 and 71C3, the chimera contains the N-terminal domain of 71C1 and the C-terminal domain of 71C3 Arabidopsis thaliana

Protein Variants

Protein Variants Comment Organism
D257E/S260L by site-directed mutagenesis Arabidopsis thaliana
D262E/N265Q by site-directed mutagenesis Arabidopsis thaliana

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0152
-
 scopoletin wild type, 50 mM Tris-HCl, pH 7.0, 14 mM 2-mercaptoethanol, 30°C Arabidopsis thaliana
0.03897
-
 scopoletin 71C1/71C3 chimera including N-terminus of 71C1 (position 1-261) and C-terminus of 71C3 (position 256-476), 50 mM Tris-HCl, pH 7.0, 14 mM 2-mercaptoethanol, 30°C Arabidopsis thaliana
0.04955
-
 scopoletin 71C1/71C3 chimera including N-terminus of 71C1 (position 1-261) and C-terminus of 71C3 (position 256-476), 50 mM Tris-HCl, pH 7.0, 14 mM 2-mercaptoethanol, 30°C Arabidopsis thaliana
0.05183
-
 scopoletin wild type, 50 mM Tris-HCl, pH 7.0, 14 mM 2-mercaptoethanol, 30°C Arabidopsis thaliana
0.06045
-
 scopoletin 71C1 double mutant, 50 mM Tris-HCl, pH 7.0, 14 mM 2-mercaptoethanol, 30°C Arabidopsis thaliana
0.2772
-
 UDP-glucose wild type, 50 mM Tris-HCl, pH 7.0, 14 mM 2-mercaptoethanol, 30°C Arabidopsis thaliana
0.2826
-
 UDP-glucose 71C1/71C3 chimera including N-terminus of 71C1 (position 1-261) and C-terminus of 71C3 (position 256-476), 50 mM Tris-HCl, pH 7.0, 14 mM 2-mercaptoethanol, 30°C Arabidopsis thaliana
0.8363
-
 UDP-glucose wild type, 50 mM Tris-HCl, pH 7.0, 14 mM 2-mercaptoethanol, 30°C Arabidopsis thaliana
1.065
-
 UDP-glucose 71C1 double mutant, 50 mM Tris-HCl, pH 7.0, 14 mM 2-mercaptoethanol, 30°C Arabidopsis thaliana

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
80000
-
 by SDS-PAGE, approximate molecular weight of wild-type enzyme and mutants Arabidopsis thaliana

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
UDP-glucose + scopoletin Arabidopsis thaliana
-
 UDP + scopolin
-
 ?

Organism

Organism UniProt Comment Textmining
Arabidopsis thaliana O82381
-
-
Arabidopsis thaliana Q9LML7
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant proteins are purified by affinity chromatography using GST-Sepharose 4B Arabidopsis thaliana

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.03
-
 71C3, 50 mM Tris-HCl, pH 7.0, 14 mM 2-mercaptoethanol, 30°C Arabidopsis thaliana
1.356
-
 71C1, 50 mM Tris-HCl, pH 7.0, 14 mM 2-mercaptoethanol, 30°C Arabidopsis thaliana

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
UDP-glucose + scopoletin
-
 Arabidopsis thaliana UDP + scopolin
-
 ?

Synonyms

Synonyms Comment Organism
71C1
-
 Arabidopsis thaliana
71C3
-
 Arabidopsis thaliana

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 assay at Arabidopsis thaliana

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.06
-
 scopoletin wild type, 50 mM Tris-HCl, pH 7.0, 14 mM 2-mercaptoethanol, 30°C Arabidopsis thaliana
1.11
-
 scopoletin 71C1/71C3 chimera including N-terminus of 71C1 (position 1-261) and C-terminus of 71C3 (position 256-476), 50 mM Tris-HCl, pH 7.0, 14 mM 2-mercaptoethanol, 30°C Arabidopsis thaliana
10.07
-
 scopoletin 71C1 double mutant, 50 mM Tris-HCl, pH 7.0, 14 mM 2-mercaptoethanol, 30°C Arabidopsis thaliana
10.92
-
 scopoletin wild type, 50 mM Tris-HCl, pH 7.0, 14 mM 2-mercaptoethanol, 30°C Arabidopsis thaliana

General Information

General Information Comment Organism
physiological function glycosyltransferases of family 1 are capable of transferring sugar moieties from activated sugar donors to a wide range of small lipophilic acceptors leading to an increase in water solubility, improved chemical stability and altered biological activity of the sugar conjugates, in plants, family 1 glycosyltransferases play an important role in metabolic homeostasis, regulation of phytohormone activities and detoxification of xenobiotics such as herbicides and pesticides Arabidopsis thaliana
physiological function glycosyltransferases of family 1 are capable of transferring sugar moieties from activated sugar donors to a wide range of small lipophilic acceptors leading to an increase in water solubility, improved chemical stability and altered biological activity of the sugar conjugates, in plants, family l glycosyltransferases play an important role in metabolic homeostasis, regulation of phytohormone activities and detoxification of xenobiotics such as herbicides and pesticides Arabidopsis thaliana

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.21
-
 UDP-glucose wild type, 50 mM Tris-HCl, pH 7.0, 14 mM 2-mercaptoethanol, 30°C Arabidopsis thaliana
1.21
-
 scopoletin wild type, 50 mM Tris-HCl, pH 7.0, 14 mM 2-mercaptoethanol, 30°C Arabidopsis thaliana
3.92
-
 UDP-glucose 71C1/71C3 chimera including N-terminus of 71C1 (position 1-261) and C-terminus of 71C3 (position 256-476) Arabidopsis thaliana
3.92
-
 UDP-glucose 71C1/71C3 chimera including N-terminus of 71C1 (position 1-261) and C-terminus of 71C3 (position 256-476), 50 mM Tris-HCl, pH 7.0, 14 mM 2-mercaptoethanol, 30°C Arabidopsis thaliana
9.45
-
 UDP-glucose 71C1 double mutant Arabidopsis thaliana
13.05
-
 UDP-glucose wild type Arabidopsis thaliana
28.48
-
 scopoletin 71C1/71C3 chimera including N-terminus of 71C1 (position 1-261) and C-terminus of 71C3 (position 256-476) Arabidopsis thaliana
28.48
-
 scopoletin 71C1/71C3 chimera including N-terminus of 71C1 (position 1-261) and C-terminus of 71C3 (position 256-476), 50 mM Tris-HCl, pH 7.0, 14 mM 2-mercaptoethanol, 30°C Arabidopsis thaliana
166.6
-
 scopoletin 71C1 double mutant Arabidopsis thaliana
210.7
-
 scopoletin wild type Arabidopsis thaliana