Dr. John D'Auria
Title: Assistant Professor
Education: Ph.D. University of Michigan, 2002; Alexander von Humboldt Postdoctoral Fellow 2003-2005, Project Leader; Max Planck Institute for Chemical Ecology, 2005-2013
Research Area: Biochemistry
Office: Chemistry-CHEM 413-D
More Info: http://www.depts.ttu.edu/chemistry/Faculty/Dauria/
Principal Research Interests
- Biochemistry and Evolution of Tropane Alkaloid Biosynthesis
- Biochemistry of Plant Specialized Metabolism
- Plant Metabolic Engineering
Tropane alkaloids represent a major class of plant-derived secondary metabolites known to occur in the Solanaceae family but are also present in the families Convolvulaceae, Proteaceae, Rhizophoraceae and Erythroxylaceae. The core defining structure of tropane alkaloids is an 8-azabicyclo[3.2.1] octane nucleus. The diversity of tropane alkaloids is achieved by elaboration of this core through different types of modifications. The genus Erythroxylum (family Erythroxylaceae) contains approximately 230 species with ranges spread throughout the tropics including South America and Madagascar.
Erythroxylum coca and Erythroxylum novogranatense are the most widely used species for the production of cocaine. Very little is known as to the biological and ecological roles that cocaine and other tropane alkaloids play in plants. Their anti-cholinergic properties argue strongly in favour of deterrent activity against herbivores. We have begun molecular and biochemical studies in order to elucidate the biochemical steps which lead to the production of tropane alkaloids in E. coca plants.
Biochemistry and Evolution of Tropane Alkaloid Biosynthesis
The terminal step in the production of cocaine or other tropane related esters is thought to be the formation of the acyl ester via the action of an acyltransferase enzyme. In the case of cocaine, this acyltransferase utilizes the substrates methylecgonine and benzoyl CoenzymeA to produce cocaine and free CoA. I have been working for several years on a plant specific family of acyltransferases commonly referred to as the BAHD acyltransferases. Thus far, more than 8 BAHD acyltransferases have been isolated from Erythroxylum coca (E. coca). Recent results show one of these BAHD members exhibits cocaine synthase activity. Members of my group have successfully developed an LC-MS based ‘realtime’ enzyme assay for cocaine synthase in order to obtain very accurate kinetic data for characterization studies. We are also using antibodies made against the whole purified protein in order to perform immunoprecipitation and immunohistochemical studies.
In addition to the study of the role of acyltransferases in E. coca, I am also actively pursuing what enzymes are involved in forming the first and second rings of the tropane core. Most theories to date suggest that the precursor compound is most likely the mono-methylated polyamine putrescine. With the aid of Dr. Christin Fellenberg, a postdoctoral fellow in my lab we are characterizing the properties of several polyamine synthases that are similar to putrescine methyltransferase and spermine/spermidine synthases. We are also interested in the origins of the benzoic acid portion of cocaine and are combining all of our tools that we have thus far developed for E. coca to develop this system as a model for benzoic acid biosynthesis.
- "The last step in cocaine biosynthesis is catalyzed by a BAHD acyltransferase."Schmidt, G., Jirschitzka, J., Porta, T., Reichelt, M., Luck, K., Pardo-Torre, J., Dolke, F., Varesio, E., Hopfgartner, G., Gershenzon, J., D'Auria, J. (2015).Plant Physiology, 16(1), 89-101. doi:10.1104/pp.114.248187
- "Influence of medium and elicitors on the production of cocaine, amino acids and phytohormones by Erythroxylum coca calli.",Docimo T, Davis AJ, Luck K, Fellenberg C, Reichelt M, Phillips M, Gershenzon J, D'Auria JC.(2015 in press) Plant Cell, Tissue and Organ Culture (PCTOC)doi: 10.1007/s11240-014-0660-8
- "Increasing the Pace of New Discoveries in Tropane Alkaloid Biosynthesis". Jan Jirschitzka, Franziska Dolke, John C. D'Auria,. In Nathalie Giglioli-Guivarc'h, editors : Advances In Botanical Research, Vol. 68,Burlington: Academic Press, 2013, pp. 39-72. ISBN: 978-0-12-408061-4
- "Selection and validation of reference genes for quantitative gene expression studies in Erythroxylum coca", Docimo, T., Schmidt, G., Luck, K., Delaney, S. K., D'Auria, J (2013).Faculty of 1000 Research,2:37. doi:10.3410/f1000research.2-37.v1.
- "Learning from nature: new approaches to the metabolic engineering of plant defense pathways", Jirschitzka, J., Mattern, D. J.,Gershenzon, J., D'Auria, J. (2013).Current Opinion in Biotechnology,24(2),320-328. doi:10.1016/j.copbio.2012.10.014.
- "The biosynthesis of hydroxycinnamoyl quinate esters and their role in the storage of cocaine in Erythroxylum coca." Pardo-Torre, J., Schmidt,G., Paetz, C., Reichelt, M., Schneider, B., Gershenzon, J.' D'Auria, J. (2013). Phytochemistry, 91, 177-186. doi:10.1016/j.phytochem.2012.09.009
- "The first step in the biosynthesis of cocaine in Erythroxylum coca: the characterization of arginine and ornithine decarboxylases.", Docimo, T., Reichelt, M., Schneider, B., Kunert, G., Gershenzon, J., D'Auria, J. ( 2012).Plant Molecular Biology, 78(6), 599-615. doi:10.1007/s11103-012-9886-1.
- "Plant tropane alkaloid biosynthesis evolved independently in the Solanaceae and Erythroxylaceae.", Jirschitzka, J., Schmidt, G., Reichelt, M., Schneider, B., Gershenzon, J., D'Auria, J. (2012). Proceedings of the National Academy of Sciences of the United States of Amercia, 109(26), 10304-10309. doi:10.1073/pnas.1200473109.
- "Contribution of CoA ligases to benzenoid biosynthesis in Petunia flowers. ", Klempien, A.; Kaminaga, Y.; Qualley, A.; Nagegowda, D. A., Widhalm, J. R., Orlova, I., Shasany, A. K., Taguchi, G., Kish, C. M., Cooper, B. R., D'Auria, J., Rhodes, D., Pichersky, E., Dudareva, N. (2012). The Plant Cell, 24,2014-2030. doi:10.1105/tpc.112.097519.
- "Evaluation of candidate reference genes for real-time quantitative PCR of plant samples using purified cDNA as template.", Phillips, M., D'Auria, J., Luck, K.; Gershenzon, J. (2009).Plant Molecular Biology Reporter, 27(3), 407-416. doi:10.1007/s11105-008-0072-1.
- "The Arabidopsis thaliana type I isopentenyl diphosphate isomerases are targeted to multiple subcellular compartments and have overlapping functions in isoprenoid biosynthesis.", Phillips, M. A.; D'Auria, J., Gershenzon, J., Pichersky, E. (2008) The Plant Cell, 20(3), 677-696. doi:10.1105/tpc.107.053926.
- "Characterization of a BAHD acyltransferase responsible for producing the green leaf volatile (Z)-3-hexen-1-yl acetate in Arabidopsis thaliana." D'Auria, J., Pichersky, E.,Schaub, A., Hansel, A., Gershenzon, J. (2007).The Plant Journal, 49(2), 194-207.
- "Identification and characterisation of the BAHD acyltransferase malonyl CoA: Anthocyanidin 5-O-glucoside-6''-O-malonyltransferase (At5MAT) in Arabidopsis thaliana."D'Auria, J., Reichelt, M., Luck, K., Svatos, A., Gershenzon, J. (2007).FEBS Letters, 581, 872-878.
- "Characterization of seed-specific benzoyloxyglucosinolate mutations in Arabidopsis thaliana." Kliebenstein, D. J., D'Auria, J., Behere, A. S., Kim, J. H., Gunderson, K. L., Breen, J. N., Lee, G., Gershenzon, J., Last, R. L., Jander, G. (2007).The Plant Journal, 51(6), 1062-1076.
- "Acyltransferases in plants: a good time to be BAHD." D'Auria, J. (2006). Current Opinion in Plant Biology, 9(3), 331-340.
- "The secondary metabolism of Arabidopsis thaliana: growing like a weed." D'Auria, J., Gershenzon, J. (2005).Current Opinion in Plant Biology, 8(3), 308-316.
- "An Arabidopsis thaliana gene for methylsalicylate biosynthesis, identified by a biochemical genomics approach, has a role in defense." Chen, F., D'Auria, J., Tholl, D., Ross, J. R., Gershenzon, J., Noel, J. P., Pichersky, E. (2003). An Arabidopsis thaliana gene for methylsalicylate biosynthesis, identified by a biochemical genomics approach, has a role in defense. The Plant Journal, 36(5), 577-588. Retrieved from ://000186532400001.
- "Biosynthesis and emission of terpenoid volatiles from Arabidopsis flowers." Chen, F., Tholl, D., D'Auria, J., Farooq, A., Pichersky, E., Gershenzon, J. (2003). Biosynthesis and emission of terpenoid volatiles from Arabidopsis flowers. The Plant Cell, 15(2), 481-494. Retrieved from ://000181008700014.