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Untersuchungen zur Biosynthese von ribosomal synthetisierten Lassopeptiden und makrozyklischen Peptidantibiotika

Subject Area Biochemistry
Term from 2010 to 2015
Project identifier Deutsche Forschungsgemeinschaft (DFG) - Project number 178860017
 
Final Report Year 2016

Final Report Abstract

This project was dedicated towards the understanding of the biochemical and structural aspects of novel class of ribosomally synthesized peptides antibiotics with a focus on bacterial lasso peptides and other macrocyclic peptides such as sactipeptides and cyclic-dipeptides. Applying a novel genome mining approach for lasso peptide discovery we were able to identify over 100 different gene clusters from a total of 87 different proteobacterial strains. Over 20 new lasso peptides were expressed in heterologous systems, purified and their specific MS-fragmentation patterns and some NMR-structures were elucidated. We have also in collaboration biochemically explored the roles of the ATP-dependent cysteine protease (protein B, associated with leader peptide release and folding) and the adenylate-forming enzyme (protein C, associated with acidic site chain activation and isopeptide bond formation) in the microcin J25 system, which represents a model system for lasso peptide maturation. In addition we discovered in this study a new 7-residues ring lasso peptides besides the well-known 8- and 9-membered ring structures and used the lasso peptide scaffold for peptide grafting for the first time. For the ribosomally synthesized sactipeptides (subtilosin A, Thurincin H and the sporulation killing factor) we explored their unique enzymatic mechanism for the formation of the thioether bond formation (post-translational modification) by SAM-radical enzymes. For the class of the ribosomally synthesized cyclic dipeptides that show the privileged 2, 5-diketopiperazine (DKP) scaffold, we explored in this study some of their modifying enzymes that are responsible for installing functional groups crucial for the biological activity of the resulting modified DKPs. The results obtained lead to important discoveries in the fascinating fields of lasso peptide structural elucidation, biosynthesis and grafting as well as to important insights into the mechanisms of post-translational modification of other ribosomally synthesized macrocyclic peptides.

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