Max J. Cryle
NHMRC Career Development Fellow
Understanding the Biosynthesis of the Glycopeptide Antibiotics
The glycopeptide antibiotics (GPAs) are a structurally complex and medically important class of peptide natural products that include the clinical antibiotics vancomycin and teicoplanin. 1 They contain a large number of non-proteinogenic amino acids and are produced by a linear non-ribosomal peptide synthetase (NRPS) machinery comprising seven modules. 2Furthermore, GPAs are extensively crosslinked late in their biosynthesis on the NRPS assembly line by the actions of a cascade of cytochrome P450 enzymes, a process which contributes to the rigidity, structural complexity and activity of these compounds. 3Due to the challenge of synthesizing GPAs, biosynthesis remains the only means of accessing GPAs for clinical use, which makes understanding the biosynthesis of GPAs of key importance.
Figure 1: The biosynthesis of the glycopeptide antibiotics as exemplified for teicoplanin. In this process, a linear heptapeptide precursor is first assembled by the non-ribosomal peptide synthetase (NRPS) machinery, which is then cyclized by the actions of 3-4 cytochrome P450 (Oxy) enzymes prior to cleavage from the NRPS and subsequent structural diversification of the peptide aglycone.
In this presentation, I will detail results from our recent studies into the enzymology of the peptide assembly line, the P450-cyclisation cascade and the interplay of these two important biosynthetic processes during GPA biosynthesis. This includes the characterization of key enzymatic processes during NRPS-mediated peptide biosynthesis (chlorination, thioesterase activity and reconstitution of peptide synthesis) 4 as well as the P450-mediated cyclization cascade (substrate specificity of P450 enzymes and cascade reconstitution). 5 Overall, our results demonstrate how selectivity during GPA biosynthesis is mediated through the careful orchestration of critical modification steps and interactions between the peptide-producing NRPS machinery and trans-modifying enzymes.
AffiliationsMonash University, Melbourne, Australia
1. E. Stegmann, H.-J. Frasch, W. Wohlleben, Curr. Opin. Microbiol. 2010, 13, 595-602.
2. R. D. Süssmuth, A. Mainz, Angew. Chem., Int. Ed. 2017, 56, 3770-3821.
3. K. Haslinger, M. Peschke, C. Brieke, E. Maximowitsch, M. J. Cryle, Nature 2015, 521, 105-109.
4. [a] M. Peschke, C. Brieke, M. Heimes, M. J. Cryle, ACS Chem. Biol. 2018, 13, 110-120;
4. [b] T. Kittila, C. Kittel, J. Tailhades, D. Butz, M. Schoppet, A. Buttner, R. J. A. Goode, R. B. Schittenhelm, K. H. van Pee, R. D. Sussmuth, W. Wohlleben, M. J. Cryle, E. Stegmann, Chem. Sci. 2017, 8, 5992-6004.
M. Schoppet, M. Peschke, A. Kirchberg, V. Wiebach, R. D. Süssmuth, E. Stegmann, M. J. Cryle, Chem. Sci. 2018, 10, 118-133.
5. [a] J. Tailhades, M. Schoppet, A. Greule, M. Peschke, C. Brieke, M. J. Cryle, Chem. Comm. 2018, 54, 2146-2149;
5. [b] M. Peschke, C. Brieke, R. J. Goode, R. B. Schittenhelm, M. J. Cryle, Biochemistry 2017, 56, 1239-1247.
Dr Max Cryle is an EMBL Australia Group leader in the Victorian Node, based in the Department of Biochemistry and Molecular Biology at Monash University and an associate investigator at the Australian Research Council Centre of Excellence in Advanced Molecular Imaging. After obtaining his PhD in chemistry from the University of Queensland in 2006, he moved to the Max Planck Institute for Medical Research in Heidelberg as a Cross Disciplinary Fellow of the Human Frontiers Science Program.
He was subsequently awarded funding from the German Research Foundation (Deutsche Forschungsgemeinschaft) to establish his own group to investigate glycopeptide antibiotic biosynthesis as part of the Emmy Noether program. In 2016 he joined EMBL Australia to continue his research into understanding the biosynthesis of important natural antibiotics and developing new antimicrobial agents. His group works at the boundary of chemistry and biology, where they apply a multidisciplinary approach including synthetic chemistry, biochemistry, structural biology and enzyme catalysis.