Two-Step One-Pot Synthesis of Pinoresinol from Eugenol in an Enzymatic Cascade

Article abstract of DOI:10.1002/cctc.201500182

The phytoestrogen pinoresinol is a high-value compound that has a protective effect against diverse health disorders, and thus is of interest for the pharmaceutical industry. Isolation of pinoresinol from plants suffers from low yields, and its chemical synthesis involves several work-up steps. In this study we devised a novel two-step one-pot enzymatic cascade combining a vanillyl-alcohol oxidase and a laccase for the production of pinoresinol from eugenol via the intermediate coniferyl alcohol. Along with the well-characterized vanillyl-alcohol oxidase from Penicillium simplicissimum used to catalyze the oxidation of eugenol, enzyme screening revealed three bacterial laccases that were appropriate for the synthesis of pinoresinol from coniferyl alcohol. The cascade was optimized regarding enzyme ratios, pH value, and the presence of organic solvents. Under optimized conditions, pinoresinol concentration achieved 4.4 mM (1.6 gl^-1), and this compound was isolated and analyzed. Increasing value: The high-value compound pinoresinol is synthesized in a two-step one-pot process combining the vanillyl-alcohol oxidase from Penicillium simplicissimum (PsVAO) and a bacterial laccase starting from the inexpensive substrate eugenol. tBME=tert-butyl methyl ether.