Development of an efficient and scalable bioprocess for the plant hormone 12-OPDA: Overcoming hurdles of nature´s biosynthesis

Abstract

Besides its native biological function as plant hormone, cis-(+)-12-oxo-phytodienoic acid (12-OPDA) serves as a metabolite for the cellular formation of (-)-jasmonic acid and also turned out to have an influence on mammalian cells. In order to make this biologically active, but at the same time very expensive natural product 12-OPDA broadly accessible for further biological and medicinal research, we developed an efficient bioprocess based on the utilization of a tailor-made whole-cell catalyst and following the principles of its biosynthesis in nature. After process optimization, the designed three-step one-pot synthesis of 12-OPDA starting from readily accessible α-linolenic acid was able to be conducted at technically relevant substrate loadings in the range of 5-20 g/L. The desired 12-OPDA was obtained with excellent conversion efficiency and by means of a developed efficient downstream-processing, this emulsifying as well as stereochemically labile biosynthetic metabolite 12-OPDA was then obtained with very high chemical purity (>99%) and enantio- and diastereomeric excess (>99% ee, 96% de) as well as neglectable by-product formation (<1%). With respect to a future technical application, we also demonstrated the scalability of the production of the whole cell-biocatalyst in a high-cell-density fermenation process.