Unser 10. Leibniz Plant Biochemistry Symposium am 7. und 8. Mai war ein großer Erfolg. Thematisch ging es in diesem Jahr um neue Methoden und Forschungsansätze der Naturstoffchemie. Die exzellenten Vorträge über Wirkstoffe…
Omanische Heilpflanze im Fokus der Phytochemie IPB-Wissenschaftler und Partner aus Dhofar haben jüngst die omanische Heilpflanze Terminalia dhofarica unter die phytochemische Lupe genommen. Die Pflanze ist reich an…
Geschmack ist vorhersagbar: Mit FlavorMiner. FlavorMiner heißt das Tool, das IPB-Chemiker und Partner aus Kolumbien jüngst entwickelt haben. Das Programm kann, basierend auf maschinellem Lernen (KI), anhand der…
Brückner, K.; Tissier, A.;High-level diterpene production by transient expression in Nicotiana benthamianaPlant Meth.946(2013)DOI: 10.1186/1746-4811-9-46
BackgroundCharacterization of plant terpene synthases is typically done by production of recombinant enzymes in Escherichia coli. This is often difficult due to solubility and codon usage issues. Furthermore, plant terpene synthases which are targeted to the plastids, such as diterpene synthases, have to be shortened in a more or less empirical approach to improve expression. We report here an optimized Agrobacterium-mediated transient expression assay in Nicotiana benthamiana for plant diterpene synthase expression and product analysis.ResultsAgrobacterium-mediated transient expression of plant diterpene synthases in N. benthamiana led to the accumulation of diterpenes within 3 days of infiltration and with a maximum at 5 days. Over 50% of the products were exported onto the leaf surface, thus considerably facilitating the analysis by reducing the complexity of the extracts. The robustness of the method was tested by expressing three different plant enzymes, cembratrien-ol synthase from Nicotiana sylvestris, casbene synthase from Ricinus communis and levopimaradiene synthase from Gingko biloba. Furthermore, co-expression of a 1-deoxy-D-xylulose-5-phosphate synthase from tomato and a geranylgeranyl diphosphate synthase from tobacco led to a 3.5-fold increase in the amount of cembratrien-ol produced, with maximum yields reaching 2500 ng/cm2.ConclusionWith this optimized method for diterpene synthase expression and product analysis, a single infiltrated leaf of N. benthamiana would be sufficient to produce quantities required for the structure elucidation of unknown diterpenes. The method will also be of general use for gene function discovery, pathway reconstitution and metabolic engineering of diterpenoid biosynthesis in plants.
Publikation
Mihály, T.; Bette, M.; Mihály, B.; Schmidt, J.; Schmidt, H.; Steinborn, D.;Synthesis, structure and characterization of adenine-based aminocarbene complexes of platinum(II)J. Organomet. Chem.73957-62(2013)DOI: 10.1016/j.jorganchem.2013.04.030
The reaction of the dinuclear platina-β-diketone [Pt2{(COMe)2H}2(μ-Cl)2] (1) with two equivalents of adenine and its methylated derivatives N6-R,9-R′Ade–H resulted in adenine-based aminocarbene platinum(II) complexes [Pt(COMe)Cl{CMe(N6-R,9-R′Ade–H)-κC,κN}] (R/R′ = Me/Me, 2; H/Me, 3; H/H, 4) whose identities were confirmed by NMR and IR spectroscopies as well as by high-resolution mass spectrometric investigations. Single-crystal X-ray diffraction analyses of complexes 2 and 4·THF revealed relatively short Pt–C and N–C bonds in the aminocarbene–platinum units, which is in accord with a substantial double bond character of these bonds. The electronic structure of these complexes will be further confirmed by DFT calculations as also the course of reaction.