Preparation of UDP-galacturonic acid using UDP-sugar pyrophosphorylase
| Autor: | Takao Ohashi, Sumihiro Hase, Nicolai Cramer, Takeshi Ishimizu |
|---|---|
| Rok vydání: | 2006 |
| Předmět: |
Magnetic Resonance Spectroscopy
Time Factors food.ingredient Pectin Biophysics Biochemistry Pyrophosphate Catalysis Pisum law.invention chemistry.chemical_compound food law Sugar Molecular Biology Nucleotide salvage Pyrophosphatase biology Peas Substrate (chemistry) Cell Biology Hydrogen-Ion Concentration Reference Standards Uridine Diphosphate Sugars biology.organism_classification Nucleotidyltransferases Recombinant Proteins Diphosphates Enzyme Activation carbohydrates (lipids) chemistry Recombinant DNA |
| Zdroj: | Analytical Biochemistry. 352:182-187 |
| ISSN: | 0003-2697 |
| DOI: | 10.1016/j.ab.2006.02.026 |
| Popis: | UDP-galacturonic acid, the activated form of galacturonic acid (GalUA), is synthesized both de novo and by salvage pathways. The UDP-GalUA pyrophosphorylase gene involved in the salvage pathway has not been identified. Here we show that UDP-sugar pyrophosphorylase from Pisum sativum with a broad specificity has UDP-GalUA pyrophosphorylase activity. The enzyme catalyzed the formation of UDP-GalUA and pyrophosphate from GalUA 1-phosphate and UTP with an equilibrium constant value of 0.24. The recombinant UDP-sugar pyrophosphorylase had optimal pH of 6.0, and the apparent K(m) values for GalUA 1-phosphate, UTP, UDP-GalUA, and pyrophosphate were 2.27, 1.15, 0.70, and 1.26 mM, respectively. In the presence of inorganic pyrophosphatase, the recombinant enzyme produced UDP-GalUA in an 84% yield (based on the GalUA 1-phosphate substrate) on a preparative scale. Thus, this UDP-sugar pyrophosphorylase is useful for the highly efficient production of UDP-GalUA for studies on pectin biosynthesis. |
| Databáze: | OpenAIRE |
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