ANTHOcyanin production PLatform using suspension cultures (ANTHOPLUS): strategies for purification and quality control

In ANTHOPLUS anthocyanins are produced in a robust new plant tissue culture system enabling the stable production of natural plant products. Our aim is to produce purified anthocyanins, of novel complexity in side chain decoration, or labelled with stable isotopes for assaying the composition of feedstock for natural colors, for bioavailability, bioefficacy and mechanistic research in experimental medicine and as analytical standards. Production of these compounds comprises the whole value-chain from synthesis (JIC), scale-up (UMB), preparative purification (IPK), assessment of toxicity and bioactivities (UNIVIE) to distribution for sales (PMC). We will focus on strategies established for the medium- and large-scale purification of anthocyanins from cell suspension cultures. Anthocyanin profiles of plant cell extracts and chemical structures of candidate molecules are analyzed by an LC-MS based approach. Selected compounds are purified by sequential flash and preparative reversed phase chromatography. The purity of the resulting fractions is validated by a combination of LC-MS and GC-MS approaches, as well as by enzyme assays. In addition, we will present our efforts on the production of “Antho-kits” with defined chemical composition, initially based on fruit juice preparations such as from blackberry, elderberry and bilberry. “Antho-kits” of those fruits will comprise the main anthocyanins of the respective fruits in their original abundance ratio after preparative separation from the remaining juice composites. These “Antho-kits” will be tested for their bioefficacy on cancerogenic cell lines and effects will be compared to those of fractions of the remaining composites and of the individual anthocyanins by our scientific partner UNIVIE