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A Cutinase from Trichoderma reesei with a Lid-Covered Active Site and Kinetic Properties of True Lipases.

Authors :
Roussel, Alain
Amara, Sawsan
Nyyssölä, Antti
Mateos-Diaz, Eduardo
Blangy, Stéphanie
Kontkanen, Hanna
Westerholm-Parvinen, Ann
Carrière, Frédéric
Cambillau, Christian
Source :
Journal of Molecular Biology. Nov2014, Vol. 426 Issue 22, p3757-3772. 16p.
Publication Year :
2014

Abstract

Cutinases belong to the α/β-hydrolase fold family of enzymes and degrade cutin and various esters, including triglycerides, phospholipids and galactolipids. Cutinases are able to degrade aggregated and soluble substrates because, in contrast with true lipases, they do not have a lid covering their catalytic machinery. We report here the structure of a cutinase from the fungus Trichoderma reesei ( Tr ) in native and inhibitor-bound conformations, along with its enzymatic characterization. A rare characteristic of Tr cutinase is its optimal activity at acidic pH. Furthermore, Tr cutinase, in contrast with classical cutinases, possesses a lid covering its active site and requires the presence of detergents for activity. In addition to the presence of the lid, the core of the Tr enzyme is very similar to other cutinase cores, with a central five-stranded β-sheet covered by helices on either side. The catalytic residues form a catalytic triad involving Ser164, His229 and Asp216 that is covered by the two N-terminal helices, which form the lid. This lid opens in the presence of surfactants, such as β-octylglucoside, and uncovers the catalytic crevice, allowing a C11Y4 phosphonate inhibitor to bind to the catalytic serine. Taken together, these results reveal Tr cutinase to be a member of a new group of lipolytic enzymes resembling cutinases but with kinetic and structural features of true lipases and a heightened specificity for long-chain triglycerides. [ABSTRACT FROM AUTHOR]

Details

Language :
English
ISSN :
00222836
Volume :
426
Issue :
22
Database :
Academic Search Index
Journal :
Journal of Molecular Biology
Publication Type :
Academic Journal
Accession number :
99232462
Full Text :
https://doi.org/10.1016/j.jmb.2014.09.003