Any feedback?
Literature summary for 2.7.7.64 extracted from
- Production of Arabidopsis thaliana UDP-sugar pyrophosphorylase by Pichia pastoris and its application in efficient UDP-Glucose and UDP-glucuronic acid synthesis (2019), Appl. Biochem. Microbiol., 55, 631-638 .
No PubMed abstract available
Application
| Application | Comment | Organism |
|---|---|---|
| synthesis | in mammals, uridine 5'-diphosphate-glucose (UDP-Glc) and uridine 5'-diphosphate-glucuronic acid (UDP-GlcA) are common building blocks of glycans and glycoconjugates. The commercial demand for these high-energy donors is increasing. To produce valuable UDP-GlcA in a cost-effective way, UDP-sugar pyrophosphorylase from Arabidopsis thaliana is constitutively expressed in Pichia pastoris and secreted into the extracellular medium. The synthesis of 4.2 g UDP-GlcA or 5.5 g UDP-Glc per liter of culture is revealed in the culture medium, without any need for purification. An anion exchange chromatography purification method for UDP-sugars is also developed. This route opens a door to large-scale production of the cheaper UDP-GlcA | Arabidopsis thaliana |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| constitutively expressed in Pichia pastoris | Arabidopsis thaliana |
| gene USP, functional UDP-sugar diphosphorylase from Arabidopsis thaliana is constitutively expressed in Pichia pastoris strain SMD1168H and secreted into the extracellular medium using the secretion signal from Saccharomyces cerevisiae alpha-factor prepro-peptide | Arabidopsis thaliana |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| additional information | functional UDP-sugar diphosphorylase from Arabidopsis thaliana, constitutively expressed in Pichia pastoris and secreted into the extracellular medium, is used for synthesis of UDP-alpha-D-glucuronate, purification of the UDP-sugar from medium by anion exchange chromatography. Purification and assay methods optimization, overview. The fermentation of Pichia pastoris strain SMD1168H-C-AtUSP is performed at 30°C and pH 7.0 throughout the procedure, 20% glucose is supplemented at a rate of 9 ml/h, and DO is maintained at over 20%. After 60 h, the cell wet weight stabilizes (a wet weight of 276.7 g/l is obtained at 96 h). The crude enzyme activity increases up to 84 h, then it is stabilized, with activity 1601 U/ml. Thus, USP activity increases in 5-l fermenter compared with that in shaken flasks. The optimal temperature is around 35°C, lower than the optimal temperature of enzymatic reaction by purified AtUSP (45°C), while the optimal pH value and metal ion conditions are close to those of the purified enzyme system | Arabidopsis thaliana |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Ca2+ | activates | Arabidopsis thaliana |  |
| Mg2+ | required, activates, preferred cation | Arabidopsis thaliana | |
| Mn2+ | activates | Arabidopsis thaliana | |
| additional information | the enzyme requires bivalent cations for activity. HvUSP accepts other bivalent cation such as Ca2+, Mn2+, and Zn2+, but prefers Mg2+ | Arabidopsis thaliana | |
| Zn2+ | activates | Arabidopsis thaliana | |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| UTP + alpha-D-glucose 1-phosphate | Arabidopsis thaliana | - |
diphosphate + UDP-alpha-D-glucose | - |
r | |
| UTP + alpha-D-glucuronate 1-phosphate | Arabidopsis thaliana | - |
diphosphate + UDP-alpha-D-glucuronate | - |
r | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Arabidopsis thaliana | Q9C5I1 | - |
- |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| UTP + alpha-D-glucose 1-phosphate | - |
Arabidopsis thaliana | diphosphate + UDP-alpha-D-glucose | - |
r | |
| UTP + alpha-D-glucuronate 1-phosphate | - |
Arabidopsis thaliana | diphosphate + UDP-alpha-D-glucuronate | - |
r | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| AtUSP | - |
Arabidopsis thaliana |
| UDP-sugar pyrophosphorylase | - |
Arabidopsis thaliana |
| USP | - |
Arabidopsis thaliana |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 35 | - |
crude recombinant enzyme | Arabidopsis thaliana |
| 45 | - |
purified recombinant enzyme | Arabidopsis thaliana |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.5 | 8.5 | - |
Arabidopsis thaliana |
Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.
Release 2023.1 (January 2023)
Legal
Curated at
Member of
