Influence of the conformational flexibility on the kinetics and dimerisation process of two Candida rugosa lipase isoenzymes

FEBS Letters

Hermoso, Juan¹  Rúa, M.Luisa²  López, Cristina²  Pernas, Marı́a A.² 

[1] Grupo de Cristalografı́a Macromolecular y Biologı́a Estructural, Instituto ‘Rocasolano’ CSIC, Serrano 119, 28006 Madrid, Spain;Area de Bioquı́mica y Biologı́a Molecular, Facultade de Ciencias de Ourense, Universidade de Vigo, As Lagoas s/n, 32004 Ourense, Spain

关键词: Lipase;    Kinetic;    Interfacial activation;    Inhibition;    Dimerization;    Structure;    Candida rugosa;    CRL;    Candida rugosa lipase;    TA;    triacetin;    E600;    diethyl p-nitrophenyl phosphate;   

DOI  :  10.1016/S0014-5793(01)02630-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We have investigated the interfacial activation process of two isoenzymes from Candida rugosa (Lip1 and Lip3) using triacetin as substrate. Kinetics were coupled to inhibition experiments in order to analyse the transition between the open and closed conformers. This process was slow, particularly for Lip1, in the absence of an interface provided by the substrate or a detergent. Dimers of Lip3 were also purified and their catalytic action was closer to that of a typical esterase. In spite of the high sequence homology between Lip1 and Lip3, small changes enhance hydrophobicity in the binding pocket of Lip3 and increase the flexibility of its flap. We postulated that these factors account for the higher tendency of Lip3 to dimerise fixing its open conformation.

【 授权许可】

Unknown