Natural products from plants have been utilized by mankind since thousands of years. Those ingredients that are related to health issues have been of special interest. However, while plants are a renewable natural resource, the secondary metabolites involved, e.g. in plant defense or signaling processes, are often of limited availability. At the same time, the demand for the scalable production of plant metabolite-based pharmaceutical drugs is growing. With increasing structural complexity, a de novo synthesis is rendered impractical: with numerous synthetic steps involved and, consequently, large amounts of waste accumulating, even the most elegant sequences will fail to be applied. Microbial production is a promising alternative that is based on inexpensive renewable feed stocks. The fast growth rates of microbes allow short production times. Moreover they perform multi-step syntheses in a “one pot” manner thereby reducing the amount of waste produced. The challenge lies in heterologous expression of whole metabolic pathways and their optimization towards natural product analogues.
The main aim of this joint project was the development of a multi-enzyme cascade for the production of strictosidine from 7-deoxyloganinic acid, a key intermediate for a vast number of plant monoterpenoid-indole-alkaloids, which are for instance used as powerful antitumor drugs (e.g., vinblastine and vincristine). Furthermore, while rather complex synthases are regularly successfully applied, reaction cascades involving synthetically highly versatile monooxygenases remain especially demanding. The monooxygenases are largely responsible for imparting structural and functional diversity to plant natural products. These NAD(P)H-dependent enzymes have a relatively low activity, a comparatively low stability, and often a low coupling efficiency between NAD(P)H consumption and product formation, leading to the formation of hydrogen peroxide and other reactive oxygen species, which limits their performance in production processes. These challenges were successfully tackled within this MoRe-Plants project: First results on a novel screening technique for the high throughput measurement of active monooxygenase concentration have already been published. Further findings on all project parts will be presented in due course.
Participating Core Groups
Prof. J. Pietruszka, Bioorganische Chemie, HHU Düsseldorf/FZ Jülich
Prof. J. Büchs, Bioverfahrenstechnik, RWTH Aachen University
Prof. U. Schwaneberg, Biotechnologie, RWTH Aachen University
Prof. V. Urlacher, Biochemie, HHU Düsseldorf
Prof B. Usadel, Biologie, RWTH Aachen University/FZ Jülich
Prof. J. Pietruszka
Bioorganische Chemie (BOC)
HHU Düsseldorf/FZ Jülich
phone: +49 (0) 2461 – 614158
01.07.2014 – 30.06.2016
MoRe-Plants is part of the NRW-Strategieprojekt BioSC and thus funded by the Ministry of Innovation, Science and Research of the German State of North Rhine-Westphalia.
Hartl, K, Denton, A, Franz-Oberdorf, K, Hoffmann, T, Spornraft, M, Usadel, B and Schwab, W (2017). Early metabolic and transcriptional variations in fruit of natural white-fruited fragaria vesca genotypes. Sci Rep 7: 45113.
Welters, T, Horn, T, Ruff, AJ, Schwaneberg, U and Buchs, J (2017). Novel technique for high throughput measurement of active monooxygenase concentration. Biotechnol Bioeng 114(4): 929-933.