Protein engineering of enzymes involved in lipid modification

Oroz‐Guinea, Isabel; Zorn, Katja; Brundiek, Henrike

doi:10.1016/b978-0-12-813167-1.00002-5

Cited by 2 publications

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References 197 publications

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“…Predictably, lipases are the most obvious choice to achieve this task as they naturally catalyze the hydrolysis of triacylglycerides (TAG) into glycerol and free FA (FFA). Moreover, this class of enzymes displays other interesting features from an industrial point of view, such as: (1) broad substrate spectrum; (2) excellent chemo‐, regio‐, and stereoselectivity; (3) high stability in harsh reaction conditions; (4) independence of cofactors; (5) enabling efficient transesterifications; and, furthermore, (6) a wide variety of lipases is commercially available (Oroz‐Guinea et al, ; Zorn et al, ). Consequently, several studies analyzing the selectivity of lipases toward different FA and their potential use for the enrichment of gondoic and erucic acids were described.…”

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confidence: 99%

Enrichment of Erucic and Gondoic Fatty Acids from Crambe and Camelina Oils Catalyzed by Geotrichum candidum Lipases I and II

Oroz‐Guinea

Zorn²,

Bornscheuer³

2019

J Americ Oil Chem Soc

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Erucic (22:1, cisΔ13) and gondoic acids (20:1, cisΔ11) are building blocks obtained from renewable sources for the oleochemical industry. Different biocatalytic strategies for the enrichment of these compounds with high recovery yields were developed in our group. Geotrichum candidum lipases (GCL) strongly discriminate against fatty acids longer than 18 carbon atoms. Thus, GCL-I and -II were investigated using hydrolysis or ethanolysis reactions with Crambe and Camelina oils. Hydrolysis was also studied using fatty acid ethyl esters (FAEE) derived from the corresponding oil. Both isoforms were highly selective; however, interesting differences were observed. Although it has been reported that GCL-I displays a higher preference toward 18 cisΔ9, which is present in the studied oils at high levels, GCL-II showed higher enrichment values during hydrolysis independent of the substrate used. Hence, enrichments of 87% (Crambe oil) and 82% (Crambe FAEE) for erucic acid and 50% (Camelina oil) and 45% (Camelina FAEE) for gondoic acid, with recovery values between 89% and 99%, were achieved. On the contrary, the best enzyme for ethanolysis was GCL-I (82% and 41% for erucic and gondoic acid, respectively). In this case, although GCL-II also displayed good enrichment and recovery levels (77% and 28%, respectively), they were lower compared to the former reactions. In both ethanolysis reactions, the FAEE fraction contained between 92% and 97% of 18 unsaturated fatty acids.

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Section: Introductionmentioning

confidence: 99%

Enrichment of Erucic and Gondoic Fatty Acids from Crambe and Camelina Oils Catalyzed by Geotrichum candidum Lipases I and II

Oroz‐Guinea

Zorn²,

Bornscheuer³

2019

J Americ Oil Chem Soc

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Enhancement of Lipase CAL‐A Selectivity by Protein Engineering for the Hydrolysis of Erucic Acid from Crambe Oil

Oroz‐Guinea

Zorn

Bornscheuer

2019

Euro J Lipid Sci & Tech

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The aim of this study is to pursue the identification and characterization of different CAL-A variants displaying higher specificity toward erucic acid than CAL-A wild type (wt). A careful analysis of the data generated from previously created site-directed saturation libraries reveals several variants that display a higher preference for the hydrolysis of p-nitrophenyl (pNP)-erucate over pNP-oleate than the wt. The best three candidates (CAL-A V238D, V238Y, and V286N) are applied in biocatalysis using both Crambe oil and ethyl ester derivatives. When acting on Crambe oil, these CAL-A variants are as efficient as CAL-A wt in terms of C22:1 enrichment and product recovery independently of the temperature (enrichment and recovery values between 70-76% and 67-79% at 37 C, and between 71-73% and 61-75% at 50 C). In contrast, hydrolysis of Crambe ethyl esters leads to substantially increased accumulations of C22:1 and recovery values (V238Y: 78% enrichment and 92% recovery; V286N: 83% enrichment and 91% recovery) when using CAL-A V238Y and CAL-A V286N compared to CAL-A wt (78% enrichment, 60% recovery) in the free fatty acid fraction. Practical Applications: This study describes the enhancement of lipase CAL-A selectivity for the isolation and recovery of erucic acid (C22:1) from plant oil or its ethyl ester derivatives. Hence, this approach could represent a more eco-friendly alternative for its application in processes where the erucic acid is used as building block, such as the production of surfactants or polymers.

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Protein engineering of enzymes involved in lipid modification

Cited by 2 publications

References 197 publications

Enrichment of Erucic and Gondoic Fatty Acids from Crambe and Camelina Oils Catalyzed by Geotrichum candidum Lipases I and II

Enrichment of Erucic and Gondoic Fatty Acids from Crambe and Camelina Oils Catalyzed by Geotrichum candidum Lipases I and II

Enhancement of Lipase CAL‐A Selectivity by Protein Engineering for the Hydrolysis of Erucic Acid from Crambe Oil

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