Principles of lipid – enzyme interactions in the limbus region of the catalytic site of Candida antarctica Lipase B
Lipases (E.C. 3.1.1.3) are ubiquitous hydrolases for the carboxyl ester bond of water-insoluble substrates such as triacylglycerols and phospholipids. Candida antarctica Lipase B (CALB) acts in aqueous as well as in low-water media, thus being of considerable biochemical significance with high inter...
Published in: | International Journal of Biological Macromolecules |
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Online Access: | http://hdl.handle.net/11566/276915 https://doi.org/10.1016/j.ijbiomac.2020.04.061 |
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ftupmarcheiris:oai:iris.univpm.it:11566/276915 2024-04-14T08:02:43+00:00 Principles of lipid – enzyme interactions in the limbus region of the catalytic site of Candida antarctica Lipase B Lucia Silvestrini Michele Cianci Silvestrini, Lucia Cianci, Michele 2020 ELETTRONICO http://hdl.handle.net/11566/276915 https://doi.org/10.1016/j.ijbiomac.2020.04.061 eng eng info:eu-repo/semantics/altIdentifier/pmid/32380114 info:eu-repo/semantics/altIdentifier/wos/WOS:000564486400009 volume:158 firstpage:358 lastpage:363 numberofpages:6 journal:INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES http://hdl.handle.net/11566/276915 doi:10.1016/j.ijbiomac.2020.04.061 info:eu-repo/semantics/altIdentifier/scopus/2-s2.0-85084323883 Lipase fatty acid/metabolism lipids/chemistry interfacial enzymology tributyrin CALB info:eu-repo/semantics/article 2020 ftupmarcheiris https://doi.org/10.1016/j.ijbiomac.2020.04.061 2024-03-21T18:27:36Z Lipases (E.C. 3.1.1.3) are ubiquitous hydrolases for the carboxyl ester bond of water-insoluble substrates such as triacylglycerols and phospholipids. Candida antarctica Lipase B (CALB) acts in aqueous as well as in low-water media, thus being of considerable biochemical significance with high interest also for its industrial applications. The hydrolysis reaction follows a two-step mechanism, or ‘interfacial activation’, with adsorption of the enzyme to a heterogeneous interface and subsequent enhancement of the lipolytic activity. Once positioned within the catalytic triad, substrates are then hydrolysed, and products released. However, the intermediate steps of substrate transfer from the lipidic-aqueous phase to the enzyme surface and then down to the catalytic site are still unclear. By inhibiting CALB with ethyl phosphonate and incubating with glyceryl tributyrate (2,3-di(butanoyloxy)propyl butanoate), the crystal structure of the lipid-enzyme complex, at 1.55 Å resolution, shows the tributyrin in the limbus region of active site. The substrate is found above the catalytic Ser, with the glycerol backbone readily pre-aligned for further processing by key interactions via an extended water network with α-helix10 and α-helix5. The findings offer new elements to elucidate the mechanism of substrate recognition, transfer and catalysis of Candida antarctica Lipase B (CALB) and lipases in general. Article in Journal/Newspaper Antarc* Antarctica Università Politecnica delle Marche: IRIS International Journal of Biological Macromolecules 158 358 363 |
institution |
Open Polar |
collection |
Università Politecnica delle Marche: IRIS |
op_collection_id |
ftupmarcheiris |
language |
English |
topic |
Lipase fatty acid/metabolism lipids/chemistry interfacial enzymology tributyrin CALB |
spellingShingle |
Lipase fatty acid/metabolism lipids/chemistry interfacial enzymology tributyrin CALB Lucia Silvestrini Michele Cianci Principles of lipid – enzyme interactions in the limbus region of the catalytic site of Candida antarctica Lipase B |
topic_facet |
Lipase fatty acid/metabolism lipids/chemistry interfacial enzymology tributyrin CALB |
description |
Lipases (E.C. 3.1.1.3) are ubiquitous hydrolases for the carboxyl ester bond of water-insoluble substrates such as triacylglycerols and phospholipids. Candida antarctica Lipase B (CALB) acts in aqueous as well as in low-water media, thus being of considerable biochemical significance with high interest also for its industrial applications. The hydrolysis reaction follows a two-step mechanism, or ‘interfacial activation’, with adsorption of the enzyme to a heterogeneous interface and subsequent enhancement of the lipolytic activity. Once positioned within the catalytic triad, substrates are then hydrolysed, and products released. However, the intermediate steps of substrate transfer from the lipidic-aqueous phase to the enzyme surface and then down to the catalytic site are still unclear. By inhibiting CALB with ethyl phosphonate and incubating with glyceryl tributyrate (2,3-di(butanoyloxy)propyl butanoate), the crystal structure of the lipid-enzyme complex, at 1.55 Å resolution, shows the tributyrin in the limbus region of active site. The substrate is found above the catalytic Ser, with the glycerol backbone readily pre-aligned for further processing by key interactions via an extended water network with α-helix10 and α-helix5. The findings offer new elements to elucidate the mechanism of substrate recognition, transfer and catalysis of Candida antarctica Lipase B (CALB) and lipases in general. |
author2 |
Silvestrini, Lucia Cianci, Michele |
format |
Article in Journal/Newspaper |
author |
Lucia Silvestrini Michele Cianci |
author_facet |
Lucia Silvestrini Michele Cianci |
author_sort |
Lucia Silvestrini |
title |
Principles of lipid – enzyme interactions in the limbus region of the catalytic site of Candida antarctica Lipase B |
title_short |
Principles of lipid – enzyme interactions in the limbus region of the catalytic site of Candida antarctica Lipase B |
title_full |
Principles of lipid – enzyme interactions in the limbus region of the catalytic site of Candida antarctica Lipase B |
title_fullStr |
Principles of lipid – enzyme interactions in the limbus region of the catalytic site of Candida antarctica Lipase B |
title_full_unstemmed |
Principles of lipid – enzyme interactions in the limbus region of the catalytic site of Candida antarctica Lipase B |
title_sort |
principles of lipid – enzyme interactions in the limbus region of the catalytic site of candida antarctica lipase b |
publishDate |
2020 |
url |
http://hdl.handle.net/11566/276915 https://doi.org/10.1016/j.ijbiomac.2020.04.061 |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_relation |
info:eu-repo/semantics/altIdentifier/pmid/32380114 info:eu-repo/semantics/altIdentifier/wos/WOS:000564486400009 volume:158 firstpage:358 lastpage:363 numberofpages:6 journal:INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES http://hdl.handle.net/11566/276915 doi:10.1016/j.ijbiomac.2020.04.061 info:eu-repo/semantics/altIdentifier/scopus/2-s2.0-85084323883 |
op_doi |
https://doi.org/10.1016/j.ijbiomac.2020.04.061 |
container_title |
International Journal of Biological Macromolecules |
container_volume |
158 |
container_start_page |
358 |
op_container_end_page |
363 |
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1796316970460643328 |