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Alteration of Chain Length Selectivity of Candida antarctica Lipase A by Semi‐Rational Design for the Enrichment of Erucic and Gondoic Fatty Acids

Biotechnological strategies using renewable materials as starting substrates are a promising alternative to traditional oleochemical processes for the isolation of different fatty acids. Among them, long chain mono‐unsaturated fatty acids are especially interesting in industrial lipid modification,...

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Autores principales: Zorn, Katja, Oroz‐Guinea, Isabel, Brundiek, Henrike, Dörr, Mark, Bornscheuer, Uwe T.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6283244/
https://www.ncbi.nlm.nih.gov/pubmed/30555288
http://dx.doi.org/10.1002/adsc.201800889
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author Zorn, Katja
Oroz‐Guinea, Isabel
Brundiek, Henrike
Dörr, Mark
Bornscheuer, Uwe T.
author_facet Zorn, Katja
Oroz‐Guinea, Isabel
Brundiek, Henrike
Dörr, Mark
Bornscheuer, Uwe T.
author_sort Zorn, Katja
collection PubMed
description Biotechnological strategies using renewable materials as starting substrates are a promising alternative to traditional oleochemical processes for the isolation of different fatty acids. Among them, long chain mono‐unsaturated fatty acids are especially interesting in industrial lipid modification, since they are precursors of several economically relevant products, including detergents, plastics and lubricants. Therefore, the aim of this study was to develop an enzymatic method in order to increase the percentage of long chain mono‐unsaturated fatty acids from Camelina and Crambe oil ethyl ester derivatives, by using selective lipases. Specifically, the focus was on the enrichment of gondoic (C20:1 cisΔ11) and erucic acid (C22:1 cisΔ13) from Camelina and Crambe oil derivatives, respectively. The pursuit of this goal entailed several steps, including: (i) the choice of a suitable lipase scaffold to serve as a protein engineering template (Candida antarctica lipase A); (ii) the identification of potential amino acid targets to disrupt the binding tunnel at the adequate location; (iii) the design, creation and high‐throughput screening of lipase mutant libraries; (iv) the study of the selectivity towards different chain length p‐nitrophenyl fatty acid esters of the best hits found, as well as the analysis of the contribution of each amino acid change and the outcome of combining several of the aforementioned residue alterations and, finally, (v) the selection and application of the most promising candidates for the fatty acid enrichment biocatalysis. As a result, enrichment of C22:1 from Crambe ethyl esters was achieved either, in the free fatty acid fraction (wt, 78%) or in the esterified fraction (variants V1, 77%; V9, 78% and V19, 74%). Concerning the enrichment of C20:1 when Camelina oil ethyl esters were used as substrate, the best variant was the single mutant V290W, which doubled its content in the esterified fraction from approximately 15% to 34%. A moderately lower increase was achieved by V9 and its two derived triple mutant variants V19 and V20 (27%). [Image: see text]
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spelling pubmed-62832442018-12-14 Alteration of Chain Length Selectivity of Candida antarctica Lipase A by Semi‐Rational Design for the Enrichment of Erucic and Gondoic Fatty Acids Zorn, Katja Oroz‐Guinea, Isabel Brundiek, Henrike Dörr, Mark Bornscheuer, Uwe T. Adv Synth Catal Full Papers Biotechnological strategies using renewable materials as starting substrates are a promising alternative to traditional oleochemical processes for the isolation of different fatty acids. Among them, long chain mono‐unsaturated fatty acids are especially interesting in industrial lipid modification, since they are precursors of several economically relevant products, including detergents, plastics and lubricants. Therefore, the aim of this study was to develop an enzymatic method in order to increase the percentage of long chain mono‐unsaturated fatty acids from Camelina and Crambe oil ethyl ester derivatives, by using selective lipases. Specifically, the focus was on the enrichment of gondoic (C20:1 cisΔ11) and erucic acid (C22:1 cisΔ13) from Camelina and Crambe oil derivatives, respectively. The pursuit of this goal entailed several steps, including: (i) the choice of a suitable lipase scaffold to serve as a protein engineering template (Candida antarctica lipase A); (ii) the identification of potential amino acid targets to disrupt the binding tunnel at the adequate location; (iii) the design, creation and high‐throughput screening of lipase mutant libraries; (iv) the study of the selectivity towards different chain length p‐nitrophenyl fatty acid esters of the best hits found, as well as the analysis of the contribution of each amino acid change and the outcome of combining several of the aforementioned residue alterations and, finally, (v) the selection and application of the most promising candidates for the fatty acid enrichment biocatalysis. As a result, enrichment of C22:1 from Crambe ethyl esters was achieved either, in the free fatty acid fraction (wt, 78%) or in the esterified fraction (variants V1, 77%; V9, 78% and V19, 74%). Concerning the enrichment of C20:1 when Camelina oil ethyl esters were used as substrate, the best variant was the single mutant V290W, which doubled its content in the esterified fraction from approximately 15% to 34%. A moderately lower increase was achieved by V9 and its two derived triple mutant variants V19 and V20 (27%). [Image: see text] John Wiley and Sons Inc. 2018-09-26 2018-11-05 /pmc/articles/PMC6283244/ /pubmed/30555288 http://dx.doi.org/10.1002/adsc.201800889 Text en © 2018 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA. This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.
spellingShingle Full Papers
Zorn, Katja
Oroz‐Guinea, Isabel
Brundiek, Henrike
Dörr, Mark
Bornscheuer, Uwe T.
Alteration of Chain Length Selectivity of Candida antarctica Lipase A by Semi‐Rational Design for the Enrichment of Erucic and Gondoic Fatty Acids
title Alteration of Chain Length Selectivity of Candida antarctica Lipase A by Semi‐Rational Design for the Enrichment of Erucic and Gondoic Fatty Acids
title_full Alteration of Chain Length Selectivity of Candida antarctica Lipase A by Semi‐Rational Design for the Enrichment of Erucic and Gondoic Fatty Acids
title_fullStr Alteration of Chain Length Selectivity of Candida antarctica Lipase A by Semi‐Rational Design for the Enrichment of Erucic and Gondoic Fatty Acids
title_full_unstemmed Alteration of Chain Length Selectivity of Candida antarctica Lipase A by Semi‐Rational Design for the Enrichment of Erucic and Gondoic Fatty Acids
title_short Alteration of Chain Length Selectivity of Candida antarctica Lipase A by Semi‐Rational Design for the Enrichment of Erucic and Gondoic Fatty Acids
title_sort alteration of chain length selectivity of candida antarctica lipase a by semi‐rational design for the enrichment of erucic and gondoic fatty acids
topic Full Papers
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6283244/
https://www.ncbi.nlm.nih.gov/pubmed/30555288
http://dx.doi.org/10.1002/adsc.201800889
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