A new method to determine enzyme activities in two-phase systems by Flow Injection Analysis (FIA)

被引：3

作者：

Becker, M ^{[1
]}

Spohn, U ^{[1
]}

机构：

[1] Univ Halle Wittenberg, Inst Biotechnol, D-06120 Halle, Germany

来源：

BIOSENSORS & BIOELECTRONICS | 1998年 / 13卷 / 7-8期

关键词：

phospholipase D; Streptomyces chromofuscus; organic solvent; log P;

D O I：

10.1016/S0956-5663(98)00064-5

中图分类号：

Q6 [生物物理学];

学科分类号：

071011 ;

摘要：

The activity of phospholipase D (PLD) from Streptomyces chromofuscus in two-phase systems consisting of a PLD containing aqueous phase and a phosphatidylcholine (PC) containing organic solvent phase was investigated. Two-phase systems with highly reproducible segmentation were generated in a continuous flow system using a capillary segmentor. Between 0.01 and 2 U/ml PLD from Streptomyces chromofuscus were detected in a water/hexane two-phase system. The linear detection range was 0.01 to 0.5 U PLD/ml. The apparent K-M of Streptomyces chromofuscus PLD in a water/hexane system was 41.43 mu M PC, the apparent V-max was 3.68 mu mole/min mg. The variation of PLD activity in different organic solvents correlated with the solvent's hydrophobicity. If anionic detergents or primary alcohols were added to the two-phase system, concentration dependent activation of the PLD was observed. (C) 1998 Elsevier Science S.A. All rights reserved.

引用

页码：925 / 930

页数：6

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