Measurement of phytase activity using tethered phytic acid as an artificial substrate: Methods development [An article from: Soil Biology and Biochemistry]
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- Language:English (Published)
- Publication Date:January 1, 2007
This digital document is a journal article from Soil Biology and Biochemistry, published by Elsevier in 2007. The article is delivered in HTML format and is available in your Amazon.com Media Library immediately after purchase. You can view it with any web browser.
The artificial chromophoric substrate analog of phytic acid, 5-O-[6-(benzoylamino)hexyl]-d-myo-inositol-1,2,3,4,6-pentakisphosphate (T-IP"5), may prove useful in measuring soil phytase activity. This chemical probe allows for direct measurement of phytase-catalyzed dephosphorylation (i.e., hydrolysis of the phosphoester bond) using high-performance liquid chromatography with UV detection. Before T-IP"5 can be used to measure phytase activity in environmental samples (soil, stream sediment, manure) refinement of the T-IP"5 probe methodology is required. Using ^3^1P nuclear magnetic resonance (NMR) spectroscopy, we identified 5-O-[6-(benzoylamino)hexyl]-d-myo-inositol-trisphosphate (T-IP"3) as the key intermediate that accumulates during phytase-catalyzed dephosphorylation of T-IP"5. An improved HPLC method for separation of reactants is also presented.
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