✦ LIBER ✦

A radiometric assay for 5-enolpyruvylshikimate-3-phosphate synthase

✍ Scribed by Eric S. Sharps

Publisher: Elsevier Science
Year: 1984
Tongue: English
Weight: 454 KB
Volume: 140
Category: Article
ISSN: 0003-2697
DOI: 10.1016/0003-2697(84)90151-9

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✦ Synopsis

A new assay for 5-enolpyruvylshikimate-3-phosphate synthase is described. This enzyme of the shikimate pathway of aromatic amino acid biosynthesis generates 5-enolpyruvylshikimate 3-phosphate and orthophosphate from phosphoenolpyruvate and shikimate 3-phosphate. The shikimate pathway is present in bacteria and plants but not in mammals. The assay employs a paper-chromatographic separation of radiolabeled substrate from product. The method is specific, is sensitive to 50 pmol of product, and is suitable for use in crude extracts of bacteria. This enzyme appears to be the primary target site of the commercial herbicide glyphosate (N-phosphonomethyl glycine). A procedure for the enzymatic synthesis of [14C]shikimate 3-phosphate from the commercially available precursor [14C]shikimic acid is also described.

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