LAUSR

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.