Alteration of Chain Length Selectivity ofCandida antarcticaLipase A by Semi‐Rational Design for the Enrichment of Erucic and Gondoic Fatty Acids
| Autor: | Henrike Brundiek, Uwe T. Bornscheuer, Katja Zorn, Isabel Oroz-Guinea, Mark Dörr |
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| Rok vydání: | 2018 |
| Předmět: |
lipid modification
0301 basic medicine chemistry.chemical_classification Candida antarctica lipase A biology long chain mono-unsaturated fatty acid enrichment Fatty acid General Chemistry Oleochemical biology.organism_classification Camelina Amino acid 03 medical and health sciences chemistry.chemical_compound 030104 developmental biology chemistry Erucic acid Biocatalysis biology.protein Candida antarctica Food science Lipid modification Lipase semi-rational protein design |
| Zdroj: | Advanced Synthesis & Catalysis |
| ISSN: | 1615-4169 1615-4150 |
| Popis: | 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 monounsaturated 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 fromCamelinaandCrambeoil ethyl ester derivatives, by using selective lipases. Specifically, the focus was on the enrichment of gondoic (C20:1cisΔ11) and erucic acid (C22:1cisΔ13) fromCamelinaandCrambeoil 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 antarcticalipase 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 lengthp-trophenyl 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 fromCrambeethyl 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 whenCamelinaoil 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 byV9 and its two derived triple mutant variants V19 and V20 (27%). |
| Databáze: | OpenAIRE |
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