Protein Biochemistry and Drug Development

The group is doing research on the metabolic pathway, food pathogens and anaerobic biocatalysis. The interdisciplinary orientation is made possible by the latest technical equipment. The junior research group is embedded into the division Food Chemistry I.

Foci of Research

Protein Biochemistry and Drug Development

  • The mevalonate-independent terpene biosynthesis and the biosynthesis of flavins: High-Throughput-Screening for active substances for new antibiotics, tuberculostatics and malaria drugs (terpene biosynthesis)
  • Concomitant study of protein/ligand-interactions by modern structure determination methods (crystallography, small-angle scattering)
  • The metabolic pathway of arginine: The control of the surrogate-marker ADMA and SDMA (asymmetric resp. symmetric dimethyl arginine) makes a regulation of the nitrogen monoxide production possible; prophylactic against coronary heart diseases, hypertension and arteriosclerosis.

Microbiology

  • Pseudomonas aeruginosa: Identified active substances can be tested directly against pathogens by in vivo inhibition studies. (Special laboratory S1 risk group 2 is available at the institute).
  • Surrogate organisms are employed for Mycobacterium tuberculosis and EHEC, for example Mycobacterium phlei.

Anaerobic Biotechnology

Mutagenesis on enoate reductase for “asymmetric bioreduction”. These catalyse trans-specific Michael-Additions with high enantio-selectivity. β-prochiral aldehydes and β-prochiral nitro compounds are interesting new substrates. 

Equipment

Two pipetting robots are available for a quick aliquotation of a 150.000 compounds encompassing substance library as well as for the preparation of assay plates for screens with a high-throughput method. Beyond that three state-of-the-art plate-reader-photometer and two dispensers are ready for use for the HTS.

Binding affinities on enzymes of the different biosynthetic pathways are measured by isothermal colorimetry.

(Almost) all working steps can be performed under exclusion of oxygen in two coy-glove-boxes with altogether 5 qm working space.

The research group has laboratories with security level S1 and S2 risk group 2 for dealings with pathogen and genetically modified microorganisms at its disposal.

For a defined and large-volume fermentation a 10 l fermenter can be run aerobic and anaerobic.

Publications

Crystal Structures of Mutant IspH Proteins Reveal a Rotation of the Substrate's Hydroxymethyl Group during Catalysis. Span, I., Gräwert, T., Bacher, A., Eisenreich, W., Groll, M.
Journal of Molecular Biology 416, 1-9 (2012)

Reverse fosmidomycin derivatives against the antimalarial drug target IspC (Dxr). Behrendt, C. T., Kunfermann, A., Illarionova, V., Matheeussen, A., Pein, M. K., Gräwert, T., Kaiser, J., Bacher, A., Eisenreich, W., Illarionov, B., Fischer, M., Maes, L., Groll, M., Kurz, T.
Journal of Medicinal Chemistry 54, 6796-6802 (2011)

Biochemistry of the non-mevalonate isoprenoid pathway. Gräwert, T., Groll, M., Rohdich, F., Bacher, A., Eisenreich, W.
Cellular and Molecular Life Sciences 68, 3797-3814 (2011)

Reductive Dehydroxylation of Allyl Alcohols by IspH Protein. Gräwert, T., Span, I., Bacher, A., Groll, M.
Angewandte Chemie, International Edition 49, 8802-8809 (2010)

Biosynthesis of Isoprenoids: Crystal Structure of the [4Fe-4S] Cluster Protein IspG. Lee, M., Gräwert, T., Quitterer, F., Rohdich, F., Eppinger, J., Eisenreich, W., Bacher, A., Groll, M.
Journal of Molecular Biology 404, 600-610 (2010)

Synthesis and Antiplasmodial Activity of Highly Active Reverse Analogues of the Antimalarial Drug Candidate Fosmidomycin. Behrendt, C.T., Kunfermann, A., Illarionova, V., Matheeussen, A., Gräwert, T., Groll, M., Rohdich, F.,Bacher, A.,Eisenreich, W., Fischer, M., Maes, L., Kurz, T.
ChemMedChem 5, 1673-1676 (2010)

Thiazolopyrimidine Inhibitors of 2-Methylerythritol 2,4-Cyclodiphosphate Synthase (IspF) from Mycobacterium tuberculosis and Plasmodium falciparum. Geist, J., Lauw, S., Illarionova, V., Illarionov, B., Fischer, M., Gräwert, T., Rohdich, F., Eisenreich, W., Kaiser, J., Groll, M., Scheurer, C., Wittlin, S., Alonso-Gómez, J., Schweizer, B., Bacher, A., Diederich, F.
ChemMedChem 5, 1092-1101 (2010)

Biosynthesis of terpenes. Probing the reaction mechanism of IspH protein by X-ray structure analysis. Gräwert, T., Span, I., Eisenreich, W., Eppinger, J., Rohdich, F., Bacher, A., Groll, M.
Proceedings of the National Academy of Sciences of the United States of America 107, 1077-1081 (2010)

Structure of Active IspH Enzyme from Escherichia coli Provides Mechanistic Insights into Substrate Reduction. Gräwert, T., Rohdich, F., Span, I., Bacher, A., Eisenreich, W., Eppinger, J., Groll, M.
Angewandte Chemie, International Edition 48, 5756-5759 (2009)

IspH Protein from Escherichia coli and IspDF protein from Campylobacter jejuni: examination and characterization. Gräwert, T.
Dissertation, München (2006)

Biosynthesis of Isoprenoids. Purification and Properties of IspG Protein from Escherichia coli. Zepeck, F., Gräwert, T., Kaiser, J., Schramek, N., Eisenreich, W., Bacher, A., Rohdich, F.
Journal of Organic Chemistry 70, 9168-9174 (2005)

Biosynthesis of isoprenoids: a bifunctional IspDF enzyme from Campylobacter jejuni. Gabrielsen, M., Rohdich, F., Eisenreich, W., Gräwert, T., Hecht, S., Bacher, A., Hunter, W. N.
European Journal of Biochemistry 271, 3028-3035 (2004)

IspH protein of Escherichia coli: studies on iron-sulfur cluster implementation and catalysis. Gräwert, T., Kaiser, J., Zepeck, F., Laupitz, R., Hecht, S., Amslinger, S., Schramek, N., Schleicher, E., Weber, S., Haslbeck, M., Buchner, J., Rieder, C., Arigoni, D., Bacher A., Eisenreich, W., Rohdich, F.
Journal of the American Chemical Society 126, 12847-12855 (2004)

Assays for inhibitors of IspH. Adam, P., Bacher, A., Eisenreich, W., Gräwert, T., Hecht, S., Rohdich, F., Zepeck, F.
Patent: DE 10302370 A1 20040805

Stereochemical studies on the making and unmaking of isopentenyl diphosphate in different biological systems. Laupitz, R., Gräwert, T., Rieder, C., Zepeck, F., Bacher, A., Arigoni, D., Rohdich, F., Eisenreich, W.
Chemistry & Biodiversity 1 (9) 1367-1376 (2004)
Erratum: Chemistry & Biodiversity 1, 1854-1854 (2004)

The deoxyxylulose phosphate pathway of isoprenoid biosynthesis: studies on the mechanisms of the reactions catalyzed by IspG and IspH protein. Rohdich, F., Zepeck, F., Adam, P., Hecht, S., Kaiser, J., Laupitz, R., Gräwert, T., Amslinger, S., Eisenreich, W., Bacher, A., Arigoni, D.
Proceedings of the National Academy of Sciences of the United States of America 100, 1586-1591 (2003)

Biosynthesis of terpenes: studies on 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase. Adam, P., Hecht, S., Eisenreich, W., Kaiser, J., Gräwert, T., Arigoni, D., Bacher, A., Rohdich, F.
Proceedings of the National Academy of Sciences of the United States of America 99, 12108-12113 (2002)