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Detection and characterization of phospholipase D by flow injection analysis.

作者信息

Becker M, Spohn U, Ulbrich-Hofmann R

机构信息

Martin-Luther-Universität Halle/Wittenberg, Institut für Biotechnologie, Halle, Germany.

出版信息

Anal Biochem. 1997 Jan 1;244(1):55-61. doi: 10.1006/abio.1996.9864.

DOI:10.1006/abio.1996.9864
PMID:9025908
Abstract

A precisely working automated system for the investigation of phospholipases D (PLDs, EC 3.1.4.4) from plant and microbial sources with flow injection analysis (FIA) has been developed. The two versions of the FIA setup described are based on the oxidation of choline liberated from phosphatidylcholine by PLD action and catalyzed by choline oxidase and the chemiluminescence detection of hydrogen peroxide produced by this reaction. The correlation between this chemiluminescence signal and the PLD activity was linear in the range between 1 and 100 mU/ml PLD. The sampling frequency was 12 samples per hour. This method was used to compare three different PLDs from cabbage and microbial sources with respect to their pH optima, temperature stability, effectors, and v/[S]-characteristics.

摘要

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