A Spectrophotometric Assay for the Transphosphatidylation Activity of Phospholipase D Enzyme
| Autor: | Masanobu Nishikawa, Tairo Hagishita, Tadashi Hatanaka |
|---|---|
| Rok vydání: | 1999 |
| Předmět: |
Biophysics
Phospholipase Biochemistry Streptomyces Substrate Specificity Nitrophenols Hydrolysis chemistry.chemical_compound Phospholipase D Streptomyces chromofuscus Molecular Biology Phospholipids chemistry.chemical_classification Chromatography Ethanol biology Chemistry Cell Biology biology.organism_classification Quantitative determination Kinetics Enzyme Spectrophotometry |
| Zdroj: | Analytical Biochemistry. 276:161-165 |
| ISSN: | 0003-2697 |
| DOI: | 10.1006/abio.1999.4353 |
| Popis: | We developed a specific spectrophotometric assay for the quantitative determination of phospholipase D-catalyzed transphosphatidylation activity. The assay measures p-nitrophenol liberated by phospholipase D-catalyzed reaction of phosphatidyl-p-nitrophenol and ethanol in an aqueous-organic emulsion system. The release of p-nitrophenol was linear to reaction time at an early stage of the reaction with phospholipase D from Streptomyces sp. In the spectrophotometric assay for the reaction with phospholipase D from Streptomyces chromofuscus, which has higher hydrolytic activity than transphosphatidylation activity, p-nitrophenol was not found. The advantages of this novel method for measuring the transphosphatidylation activity of phospholipase D are that (i) it does not use radioactive compounds, (ii) it can measure the initial velocity of the reaction, and (iii) it is rapid, easy, and accurate to perform. |
| Databáze: | OpenAIRE |
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