Process Biochemistry, Vol.88, 22-30, 2020
A whole-cell process for the production of epsilon-caprolactone in aqueous media
epsilon-Caprolactone is an industrially important intermediate produced in multi-10,000 ton scale annually with broad applications. We report on a whole-cell biocatalytic conversion of cyclohexanol to epsilon-caprolactone using the combination of alcohol dehydrogenase (ADH) with two stability-improved variants (QM and M15) of the Baeyer-Villiger monooxygenase CHMO with a special focus on process development at the 200 mM scale. Influence of parameters such as volumetric mass transfer co-efficient, stirrer speed and catalytic loading (amount of E. coli whole-cells expressing ADH and CHMO) on the process efficiency were studied and optimised. This resulted in over 98% conversion, a product titer of 20 g L-1 and an isolated product amount of 9.1 g (80%). This corresponds to a space-time yield of 1.1 g L-1 h(-1) and a reaction yield (mole of product per mole substrate) of 0.9. Comparing the two CHMO variants a significant difference in catalytic yield (weight of product to weight of catalyst; 0.6 vs 0.3) was observed without any inherent changes in the process. Hence, the reported process can accommodate in the future improved variants of the CHMO.
Keywords:Alcohol dehydrogenase;Cyclohexanone monooxygenase;epsilon-caprolactone synthesis;Process optimisation;Whole-cell biocatalysis