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oai:www.ideals.illinois.edu:2142/121960 2024-03-01 col_2142_5131 col_2142_14789 com_2142_5130 com_2142_14788 com_2142_8903

van der Donk, Wilfred A. van der Donk, Wilfred A. Silverman, Scott K. Mitchell, Douglas A. Zhao, Huimin 2023-12 application/pdf en text Submission original under an indefinite embargo labeled 'Open Access'. The submission was exported from vireo on 2024-03-01 without embargo terms The student, Tung Le, accepted the attached license on 2023-10-19 at 11:29. The student, Tung Le, submitted this Dissertation for approval on 2023-10-19 at 11:38. This Dissertation was approved for publication on 2023-10-24 at 16:56. DSpace SAF Submission Ingestion Package generated from Vireo submission #19865 on 2024-03-01 at 13:13:55 Application of a substrate-tolerant lanthipeptide synthetase in bioengineering peptide natural products Le, Tung Thanh Lanthipeptides Enzymatic Promiscuity Bio-engineering Ripps 2023-10-24 Lanthipeptides constitute a diverse class of peptide natural products defined by the presence of lanthionine residue(s). These peptides belong to a larger family of natural products called RiPPs, which stands for ribosomally synthesized and post-translationally modified peptides. Like many RiPPs classes, the enzymes involved in the biosynthesis of lanthipeptides demonstrate relatively high substrate tolerance, a feature which, combined with the ribosomal origin of the peptide substrates, facilitates bioengineering efforts of these peptides via evolution techniques. This dissertation is thematically built around an intriguing example of extreme catalytic promiscuity in lanthipeptides biosynthesis, the prochlorosin system. The journey starts with a discussion of why and how nature has possibly evolved the prochlorosins to favor diversity-oriented biosynthesis, followed by an overview of high-throughput screening methods that have been applied to lanthipeptides. In chapter 2, the biosynthesis of prochlorosins is shown to be under substrate control such that the site-selectivity of lanthionine formation is heavily influenced by the substrates’ primary sequences. This feature opens up an opportunity for the construction of lanthipeptide libraries which is realized in chapter 3, in which an in-liposome directed evolution experiment is described in order to discover lanthipeptides with improved bioactivity. Finally, in chapter 4, the prochlorosin biosynthetic machinery is combined with an enzyme from a distant RiPP class in order to create hybrid structures that have potentials for higher stability, affinity, or adaptability for high-throughput screening experiments. Text eng https://hdl.handle.net/2142/121960 Copyright 2023 Tung Le Ph.D. Dissertation Chemistry University of Illinois at Urbana-Champaign Chemistry