Open and closed states of Candida antarctica lipase B: protonation and the mechanism of interfacial activation1
Lipases (EC 3.1.1.3) are ubiquitous hydrolases for the carboxyl ester bond of water-insoluble substrates, such as triacylglycerols, phospholipids, and other insoluble substrates, acting in aqueous as well as in low-water media, thus being of considerable physiological significance with high interest...
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ftpubmed:oai:pubmedcentral.nih.gov:4655990 2023-05-15T13:40:01+02:00 Open and closed states of Candida antarctica lipase B: protonation and the mechanism of interfacial activation1 Stauch, Benjamin Fisher, Stuart J. Cianci, Michele 2015-12 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4655990/ http://www.ncbi.nlm.nih.gov/pubmed/26447231 https://doi.org/10.1194/jlr.M063388 en eng The American Society for Biochemistry and Molecular Biology http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4655990/ http://www.ncbi.nlm.nih.gov/pubmed/26447231 http://dx.doi.org/10.1194/jlr.M063388 Copyright © 2015 by the American Society for Biochemistry and Molecular Biology, Inc. http://creativecommons.org/licenses/by/3.0/ Author’s Choice—Final version free via Creative Commons CC-BY license. CC-BY Research Articles Text 2015 ftpubmed https://doi.org/10.1194/jlr.M063388 2015-12-06T01:42:41Z Lipases (EC 3.1.1.3) are ubiquitous hydrolases for the carboxyl ester bond of water-insoluble substrates, such as triacylglycerols, phospholipids, and other insoluble substrates, acting in aqueous as well as in low-water media, thus being of considerable physiological significance with high interest also for their 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. Among lipases, Candida antarctica lipase B (CALB) has never shown any significant interfacial activation, and a closed conformation of CALB has never been reported, leading to the conclusion that its behavior was due to the absence of a lid regulating the access to the active site. The lid open and closed conformations and their protonation states are observed in the crystal structure of CALB at 0.91 Å resolution. Having the open and closed states at atomic resolution allows relating protonation to the conformation, indicating the role of Asp145 and Lys290 in the conformation alteration. The findings explain the lack of interfacial activation of CALB and offer new elements to elucidate this mechanism, with the consequent implications for the catalytic properties and classification of lipases. Text Antarc* Antarctica PubMed Central (PMC) Journal of Lipid Research 56 12 2348 2358 |
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Research Articles Stauch, Benjamin Fisher, Stuart J. Cianci, Michele Open and closed states of Candida antarctica lipase B: protonation and the mechanism of interfacial activation1 |
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Research Articles |
description |
Lipases (EC 3.1.1.3) are ubiquitous hydrolases for the carboxyl ester bond of water-insoluble substrates, such as triacylglycerols, phospholipids, and other insoluble substrates, acting in aqueous as well as in low-water media, thus being of considerable physiological significance with high interest also for their 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. Among lipases, Candida antarctica lipase B (CALB) has never shown any significant interfacial activation, and a closed conformation of CALB has never been reported, leading to the conclusion that its behavior was due to the absence of a lid regulating the access to the active site. The lid open and closed conformations and their protonation states are observed in the crystal structure of CALB at 0.91 Å resolution. Having the open and closed states at atomic resolution allows relating protonation to the conformation, indicating the role of Asp145 and Lys290 in the conformation alteration. The findings explain the lack of interfacial activation of CALB and offer new elements to elucidate this mechanism, with the consequent implications for the catalytic properties and classification of lipases. |
format |
Text |
author |
Stauch, Benjamin Fisher, Stuart J. Cianci, Michele |
author_facet |
Stauch, Benjamin Fisher, Stuart J. Cianci, Michele |
author_sort |
Stauch, Benjamin |
title |
Open and closed states of Candida antarctica lipase B: protonation and the mechanism of interfacial activation1 |
title_short |
Open and closed states of Candida antarctica lipase B: protonation and the mechanism of interfacial activation1 |
title_full |
Open and closed states of Candida antarctica lipase B: protonation and the mechanism of interfacial activation1 |
title_fullStr |
Open and closed states of Candida antarctica lipase B: protonation and the mechanism of interfacial activation1 |
title_full_unstemmed |
Open and closed states of Candida antarctica lipase B: protonation and the mechanism of interfacial activation1 |
title_sort |
open and closed states of candida antarctica lipase b: protonation and the mechanism of interfacial activation1 |
publisher |
The American Society for Biochemistry and Molecular Biology |
publishDate |
2015 |
url |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4655990/ http://www.ncbi.nlm.nih.gov/pubmed/26447231 https://doi.org/10.1194/jlr.M063388 |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_relation |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4655990/ http://www.ncbi.nlm.nih.gov/pubmed/26447231 http://dx.doi.org/10.1194/jlr.M063388 |
op_rights |
Copyright © 2015 by the American Society for Biochemistry and Molecular Biology, Inc. http://creativecommons.org/licenses/by/3.0/ Author’s Choice—Final version free via Creative Commons CC-BY license. |
op_rightsnorm |
CC-BY |
op_doi |
https://doi.org/10.1194/jlr.M063388 |
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Journal of Lipid Research |
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56 |
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12 |
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2348 |
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2358 |
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1766126868588658688 |