A Cutinase from Trichoderma reesei with a lid-covered active site and kinetic properties of true lipases

A. Roussel, S. Amara, Antti Nyyssölä, E. Mateos-Diaz, S. Blangy, Hanna Kontkanen, Ann Westerholm-Parvinen, F. Carriere, C. Cambillau

Research output: Contribution to journalArticleScientificpeer-review

19 Citations (Scopus)

Abstract

Cutinases belong to the a/ß-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 ß-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 ß-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.
Original languageEnglish
Pages (from-to)3757-3772
JournalJournal of Molecular Biology
Volume426
Issue number22
DOIs
Publication statusPublished - 2014
MoE publication typeA1 Journal article-refereed

Fingerprint

Trichoderma
Lipase
Catalytic Domain
Triglycerides
Enzymes
Galactolipids
cutinase
Organophosphonates
Hydrolases
Surface-Active Agents
Detergents
Serine
Phospholipids
Esters
Fungi

Keywords

  • cutinase
  • lipase
  • Trichoderma reesei
  • X-ray structure
  • interfacial activation

Cite this

Roussel, A. ; Amara, S. ; Nyyssölä, Antti ; Mateos-Diaz, E. ; Blangy, S. ; Kontkanen, Hanna ; Westerholm-Parvinen, Ann ; Carriere, F. ; Cambillau, C. / A Cutinase from Trichoderma reesei with a lid-covered active site and kinetic properties of true lipases. In: Journal of Molecular Biology. 2014 ; Vol. 426, No. 22. pp. 3757-3772.
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abstract = "Cutinases belong to the a/{\ss}-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 {\ss}-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 {\ss}-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.",
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A Cutinase from Trichoderma reesei with a lid-covered active site and kinetic properties of true lipases. / Roussel, A.; Amara, S.; Nyyssölä, Antti; Mateos-Diaz, E.; Blangy, S.; Kontkanen, Hanna; Westerholm-Parvinen, Ann; Carriere, F.; Cambillau, C.

In: Journal of Molecular Biology, Vol. 426, No. 22, 2014, p. 3757-3772.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - A Cutinase from Trichoderma reesei with a lid-covered active site and kinetic properties of true lipases

AU - Roussel, A.

AU - Amara, S.

AU - Nyyssölä, Antti

AU - Mateos-Diaz, E.

AU - Blangy, S.

AU - Kontkanen, Hanna

AU - Westerholm-Parvinen, Ann

AU - Carriere, F.

AU - Cambillau, C.

PY - 2014

Y1 - 2014

N2 - Cutinases belong to the a/ß-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 ß-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 ß-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.

AB - Cutinases belong to the a/ß-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 ß-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 ß-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.

KW - cutinase

KW - lipase

KW - Trichoderma reesei

KW - X-ray structure

KW - interfacial activation

U2 - 10.1016/j.jmb.2014.09.003

DO - 10.1016/j.jmb.2014.09.003

M3 - Article

VL - 426

SP - 3757

EP - 3772

JO - Journal of Molecular Biology

JF - Journal of Molecular Biology

SN - 0022-2836

IS - 22

ER -