Efficient Enzymatic Cyclization of Disulfide-Rich Peptides by Using Peptide Ligases |
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Authors: | Marcel Schmidt Dr Yen-Hua Huang Dr Eduardo F Texeira?de?Oliveira Dr Ana Toplak Dr Hein J Wijma Prof?Dr Dick B Janssen Prof?Dr Jan H van?Maarseveen Prof?Dr David J Craik Dr Timo Nuijens |
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Affiliation: | 1. EnzyPep B.V., Brightlands Campus, Urmonderbaan 22, 6167 RD Geleen, The Netherlands;2. Institute for Molecular Bioscience, The University of Queensland, Brisbane, Queensland, 4072 Australia;3. Groningen Biomolecular Science and Biotechnology Institute, University of Groningen, Nijenborgh 4, 9747 AG Groningen, The Netherlands;4. Van 't Hoff Institute for Molecular Sciences (HIMS), University of Amsterdam, Science Park 904, 1098 XH Amsterdam, The Netherlands |
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Abstract: | Disulfide-rich macrocyclic peptides—cyclotides, for example—represent a promising class of molecules with potential therapeutic use. Despite their potential their efficient synthesis at large scale still represents a major challenge. Here we report new chemoenzymatic strategies using peptide ligase variants—inter alia, omniligase-1—for the efficient and scalable one-pot cyclization and folding of the native cyclotides MCoTI-II, kalata B1 and variants thereof, as well as of the θ-defensin RTD-1. The synthesis of the kB1 variant T20K was successfully demonstrated at multi-gram scale. The existence of several ligation sites for each macrocycle makes this approach highly flexible and facilitates both the larger-scale manufacture and the engineering of bioactive, grafted cyclotide variants, therefore clearly offering a valuable and powerful extension of the existing toolbox of enzymes for peptide head-to-tail cyclization. |
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Keywords: | chemoenzymatic peptide synthesis (CEPS) cyclotides enzyme catalysis macrocycles omniligase-1 |
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