Biocatalytic enantiomeric resolution of l-menthol from an eight isomeric menthol mixture through transesterification

The four diastereomers of menthol and their enantiomers, namely dl-menthol, dl-neomenthol, dl-neoisomenthol and dl-isomenthol, were synthesised by the hydrogenation of thymol to yield an eight isomer liquid menthol. A suitably selective lipase was sought to preferentially esterify l-menthol in organic solvent, hence simplifying separation from the diasteromeric mix through distillation.From an initial screen a commercial Pseudomonas fluorescens lipase (Amano AK) was selected, and vinyl acetate was chosen as a suitable irreversible acyl donor for transesterification. The reaction reagent ratios were optimised, and an enantiomeric excess (ee) of l-menthol of greater than 95% was reproducibly achievable at a conversion of 30% dl-menthol (0.68 M) at ≤50 °C. On the basis of the composition of liquid menthol the reaction had a diastereomeric excess (de) of 82%. The enzyme was recycled 150 times in 5 ml batch reactions using liquid menthol and achieving an overall yield of 184.3 g dl-menthol/g commercial enzyme preparation. The by-product acetic acid, formed by hydrolysis of menthyl acetate, was found to cause a high degree of enzyme inactivation.The resolution reaction was scaled up 400 fold to 2 L and the enzyme recycled 38 times with an average conversion of the available l-menthol of 59% and a volumetric productivity of 1.2 g/L/h.

Graphical abstractFigure optionsDownload full-size imageDownload as PowerPoint slideHighlights► Faced with the isolation of eight diastereomers with similar chemical properties we used an enzyme to selectively transform the desired isomer (l-menthol) by esterification. ► The esterified menthol could then easily be separated from the mixture by physical means, such as distillation. ► The reaction was scaled up to 2 L and used a commercial enzyme that is available in industrial quantities.