Production and recovery of limonene-1,2-diol and simultaneous resolution of a diastereomeric mixture of limonene-1,2-epoxide with whole cells of Rhodococcus erythropolis DCL14

Cells of Rhodococcus erythropolis DCL14 present a limonene epoxide hydrolas e activity when grown on terpenes, which enables them to convert cis-limone ne-1,2-epoxide to limonene-1,2-diol. Trans-limonene-1,2-epoxide is only con verted when no cis is present. An organic/aqueous system was developed to o vercome the low solubility and instability of limonene-1,2-epoxide in the a queous phase. The presence of the organic solvent allowed high epoxide conc entrations to be used which resulted in high limonene-1,2-diol production r ates. Relatively cheap solvents were tested without any significant loss of epoxide hydrolase activity. Using a 500 mi fed-batch mechanically stirred reactor it was possible to produce 197.2 g of diol per g of protein and to obtain a yield of 98.2% and 67.9% for the trans-epoxide and the diol, respe ctively. Production of 72.4 g of diol per g of protein was obtained using a magnetically stirred reactor with an external loop for product separation. In this case, trans-epoxide and diol yields were 98.5% and 94.1%, respecti vely. A downstream process, based both on the preference of the substrate for org anic solvents and that of the product for the aqueous phase, allowed the re covery of limonene-1,2-diol, as well as of the trans-epoxide, with a purity higher than 99%.