Document Detail


A facile enzymatic synthesis of uridine diphospho-[14C]galacturonic acid.
MedLine Citation:
PMID:  10805536     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
S S Basu; G D Dotson; C R Raetz
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Analytical biochemistry     Volume:  280     ISSN:  0003-2697     ISO Abbreviation:  Anal. Biochem.     Publication Date:  2000 Apr 
Date Detail:
Created Date:  2000-06-30     Completed Date:  2000-06-30     Revised Date:  2011-04-25    
Medline Journal Info:
Nlm Unique ID:  0370535     Medline TA:  Anal Biochem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  173-7     Citation Subset:  IM    
Affiliation:
Department of Biochemistry, Duke University Medical Center, Durham, North Carolina 27710, USA.
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MeSH Terms
Descriptor/Qualifier:
Carbon Radioisotopes
Catalase / chemistry*
Galactose Oxidase / chemistry*
Magnetic Resonance Spectroscopy
Uridine Diphosphate Sugars / chemical synthesis*
Grant Support
ID/Acronym/Agency:
R37 GM051796-05/GM/NIGMS NIH HHS; R37-GM-51796/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Carbon Radioisotopes; 0/Uridine Diphosphate Sugars; 50722-58-2/UDP-galacturonic acid; EC 1.1.3.9/Galactose Oxidase; EC 1.11.1.6/Catalase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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