An active-site titration method for lipases

被引：53

作者：

Rotticci, D

Norin, T

Hult, K ^{[1
]}

Martinelle, M

机构：

[1] Royal Inst Technol, Dept Biotechnol, SE-10044 Stockholm, Sweden

[2] Royal Inst Technol, Dept Chem Organ Chem, SE-10044 Stockholm, Sweden

来源：

BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR AND CELL BIOLOGY OF LIPIDS | 2000年 / 1483卷 / 01期

关键词：

concentration; immobilized lipase; aqueous medium; organic medium; interfacial activation; detergent; inhibitor;

D O I：

10.1016/S1388-1981(99)00168-7

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A method for active-site titration of lipases has been developed based on irreversible inhibition by methyl p-nitrophenyl n-hexylphosphonate. This method was applied to five lipases displaying from minor to pronounced interfacial activation. Soluble and immobilized lipases were successfully titrated in aqueous media. A low concentration of sodium dodecyl sulfate was needed for lipases displaying pronounced interfacial activation. The carrier of some of the immobilized preparations adsorbed part of the produced p-nitrophenolate, This problem could be solved by extracting the p-nitrophenolate after inhibition. The method was extended to apolar organic solvents in the case of immobilized lipase preparations. (C) 2000 Elsevier Science B.V. All rights reserved.

引用

页码：132 / 140

页数：9

共 21 条

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