A spectrophotometric assay for the transphosphatidylation activity of phospholipase D enzyme

被引：25

作者：

Hagishita, T ^{[1
]}

Nishikawa, M ^{[1
]}

Hatanaka, T ^{[1
]}

机构：

[1] Res Inst Biol Sci, Okayama 7161241, Japan

来源：

ANALYTICAL BIOCHEMISTRY | 1999年 / 276卷 / 02期

关键词：

phospholipase D; assay method; spectrophotometry; p-nitrophenol;

D O I：

10.1006/abio.1999.4353

中图分类号：

Q5 [生物化学];

学科分类号：

071010 [生物化学与分子生物学]; 081704 [应用化学];

摘要：

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. (C) 1999 Academic Press.

引用

页码：161 / 165

页数：5

共 12 条

[1]

CARREA G, 1995, BIOCHIM BIOPHYS ACTA, V1225, P273

[2]

Using phospholipases for phospholipid modification [J].