دورية
A Facile Enzymatic Synthesis of Uridine Diphospho-[14C]galacturonic Acid
| العنوان: | A Facile Enzymatic Synthesis of Uridine Diphospho-[14C]galacturonic Acid |
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| المؤلفون: | Basu, Shib Sankar, Dotson, Garry D., Raetz, Christian R. H. |
| المصدر: | Analytical Biochemistry; April 10, 2000, Vol. 280 Issue: 1 p173-177, 5p |
| مستخلص: | Galacturonic acid (GalA) is a major component of plant cell-wall-derived pectins. It can be also found in the cell-surface polysaccharides of different microorganisms, including several symbiotic and pathogenic bacteria. Uridine diphosphogalacturonic acid (UDP-GalA) is a likely donor for GalA during the biosynthesis of these polysaccharides. A highly efficient, yet simple, method is presented for generating and purifying UDP-[14C]GalA. Commercially available UDP-[14C]galactose was quantitatively oxidized (>95% conversion) to UDP-[14C]GalA in the presence of high levels of galactose oxidase and catalase, at prolonged incubation times. Following this one-step enzymatic oxidation, UDP-[14C]GalA was purified using a polyethyleneimine cellulose column with a single-step 1 M NaCl elution. The authenticity of the purified UDP-[14C]GalA was verified by its relative mobility on thin-layer chromatograms, analysis of its chemical hydrolysis products, and 1H NMR spectroscopy. Our yield of >90% is much higher than by previously described methods. The method may serve as a prototype for the preparation of other radiolabeled uronic acids and their nucleotide derivatives. Copyright 2000 Academic Press. |
| قاعدة البيانات: | Supplemental Index |
Full Text Finder | تدمد: | 00032697 10960309 |
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| DOI: | 10.1006/abio.2000.4479 |