Chemical and enzymatic macrocyclisation of antibiotic peptides Yaoyu Ding, Gustavo Perez-Ortiz, Sarah M. Barry King's College London, UK Antimicrobial resistance is a rising global concern, it is important to speed up the discovery of novel antibiotics. Compared to other natural products inhibitors, the non-ribosomal peptide rufomycins have outstanding anti- mycobacterial activity by inhibiting bacteria enzyme ClpC1 proteolysis in M. tuberculosis and M. abscessus . 1 The rufomycin biosynthetic gene cluster has been identified from two strains. 2,3 The cyclic precursor is released through macrocyclisation by RufT thioesterase (TE) domain. The molecule is further functionalised by two cytochrome P450s, which were proved active in vitro bioassay. 4 Our goal is to combine peptide synthesis with biocatalysis and also chemical cyclisation to generate derivatives of rufomycin for future structure - activity studies. Herein, we used Solid Phase Peptide Synthesis (SPPS) to assemble linear peptide precursor and developed a chemoenzymatic synthesis employing the RufT-TE domain to cyclise and afford rufomycin analogues. Alternatively, we also developed an efficient and green cyclisation method as an in-situ reaction to give cyclic peptide on a preparative scale within 10mins. Notably, it can also cyclise peptides that cannot be achieved through enzymatic reaction. We diversify peptide building blocks through SPPS, then either chemically or enzymatically macro cyclise peptide (Figure.1). Furthermore, one-pot style enzyme cascade reaction with other tailoring enzymes to modify this scaffold will be investigated in future.
References 1. Han J, Liu X, Zhang L, Quinn RJ, Feng Y. Nat Prod Rep . 2022;39(1):77-89.Ma J, Huang H, Xie Y, et al. Nat Commun . 2017;8(1).Tomita H, Katsuyama Y, Minami H, Ohnishi Y. J Biol Chem . 2017;292(38):15859-15869.Perez Ortiz G, Sidda JD, De Los Santos ELC, Hubert CB, Barry SM. Chem Commun . 2021;57(89):11795-11798.
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