Plant Cell Biology 2022

Cinnamom belongs to the most ancient genera of flowering plants and existed already at the time, before Pangaea broke up. Thus, it is found on almost all continents and has split up into numerous species that are very hard to identify. As aromatic plant, Asiatic Cinnamom species have been used for several thousands of years, for different purposes (as spice, as ailment, as perfume). The chemical reason for the aroma are mostly phenylpropanoids (the precursors for lignin and derivatives thereof), but also terpenoids such as camphor or borneol. The commercial value is quite variable, for instance Cassia Cinnamom (formerly C. aromaticum or C. cassia) is much cheaper than Ceylon Cinnamom (C. verum). In addition, several Southeast Asian species such as Saigon Cinnamom (C. burmannii) are traded as well and used for medicinal purposes. Some species contain high levels of coumarin, which is genotoxic and therefore problematic if ingested at higher quantities. This problem has become accentuated by a new trend for Cinnamom capsules that are used to cure type II diabetes and contain high concentrations of often undefined cinnamoms. We try to develop assays to detect coumarin-rich species in such products and have established a collection of cinnamoms in our Botanical Garden. The identity of our reference plants is key to any approach to authenticate food products by genetic barcoding. However, we have learnt that many of our species, which we got from other reputed Botanical Gardens or from commercial vendors, turned out to be not the species declared. The project will use a combination of genetic barcoding, molecular phylogeny, and phenotypic analysis to shed light into the dark. Poster Masterarbeit Claudia Swoboda . Scriptum with protocols .

Microfluidic Bioreactor

During five years of cooperation with the Institute for Mikrostructure Technology (IMT) we developed this microfluidic bioreactor. In frame of a project funded by the Federal Ministry of Research we want to generate with this approch valuable compounds.
Valuable medically active plant compounds.

 

Plants produce around one million specific secondary metabolites. Many of those function to steer the interaction with other organisms and therefore many of those are pharmaceutically active. Technical synthesis of these valuable and costly compounds is often challenging or even impossible, such that they have to be extracted and purified from their natural source. The underlying metabolic pathways are complex and usually require interaction of different cell types till the compound is stored in specialised, often individual excretory cells. This renders extraction cumbersome and inefficient. Moreover, many of these plants are rare and endangered. For instance, the Pacific Yew was brought to the verge of extinction by the discovery that Theo compound taxol found in its bark can cure cancers. Biotechnological approaches would provide more sustainable alternatives.

 

Molecular Farming – Potential and Limitations.

 

Especially in situations, where small amounts of a costly product have to be produced, Green Molecular Farming turns out to excel other systems such as transgenic animals or microorganisms. Since the system of production is closed, the controversial issue of GMO spread can be circumvented. The focus, at present, is still on protein based compounds. Synthesis of secondary metabolites still plays only a marginal role. This is not caused by a lack of interest – the synthesis of the anticancer compound vinblastine in cell lines of Catharanthus roseus has been pursued over half a century by now with only modest outcome. A limitation for Molecular Farming has been the forementioned compartmentalisation of secondary metabolism to different cell types, what is difficult tob e mimicked in a bioreactor. The production of valuable plant secondary compounds requires that cell types providing different metabolic steps are coupled by a flux.

 

Our new approach – modular microfluidics.

 

Our idea starts exactly from here. Basically, we want to simulate a plant tissue technically. This is not achieved by conventional fermenters, but by a microfluidic system that provides a metabolic flux between different cell types that carry individual metabolic steps. The product is then measured in the flowthrough. By the modular organisation allows for different combinations to produced different compounds (even derivatives that would not naturally occur in the plant). A metabolic module consists of cells, where by genetic engineering specific metabolic key enzymes are overexpressed. As proof-of-principle we investigate the flavonoid-/stilbene pathway (synthesis of the potent antitumour compound quercetine-aglycon) of grapevine, the alkaloid pathway of tobacco (synthesis of the anti-Alzheimer compound nornicotine), and the alkaloid pathway of the Indian Wintercherry Withania (synthesis of withanolides with high potential against Parkinson).

 

Where we are

In cooperation with the group of the lab of Dr. Guber, Institute for Mikrostructure Technology (KIT-CN) we developed over several years a microfluidic biofermenter that is now submitted for patenting. This system allows to cultivate the plant model tobacco BY-2 over longer periods. The system was optimised to maintain biological functionality of plant cells and to harvest in the flowthrough different secondary metabolites.