Prof. Dr. Helge Bode, Max Planck Institute for terrestrial Microbiology, Germany

NRPS engineering to build new molecules and assign enzyme function

In the past 10 years we have developed several ways to modify non-ribosomal peptide synthetases (NRPS), which allows us to make novel linear and cyclic (depsi) peptides containing unnatural, hydroxylated, N-methylated and other unusual building blocks. With the recent XUT technology that allows the fusion of different NRPS parts within the thiolation domain, we could also generate hybrids or NRPS and polyketide synthases (PKS), since the thiolation domain is present in both megasynthases.

Recently, we could also use NRPS engineering to (i) assign the function of accessory genes encoding enzymes like methyltransferases and hydroxylases modifying the amino acids accepted by the associated NRPS, and (ii) to assign the function of adenylation domains with unknown substrate specificity. Furthermore, we could apply self-spicing inteins to covalently connect small NRPS fragments into functional multimodular NRPS systems and we could apply NRPS fragments for the in vitro production of different peptides from multiple individually produced and SYNZIP-modified NRPS parts.