Titlebook: Non-Ribosomal Peptide Biosynthesis and Engineering; Methods and Protocol Michael Burkart,Fumihiro Ishikawa Book 2023 The Editor(s) (if appl

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A Practical Guideline to Engineering Nonribosomal Peptide Synthetasessult of nearly one decade of NRPS engineering in the Bode lab, we provide valuable insights into the strategies we have developed during this time for the successful engineering and cloning of these fascinating molecular machines.

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In Vitro Biochemical Characterization of Excised Macrocyclizing Thioesterase Domains from Non-ribosodes substrate required for characterization of the TE. In this method, we describe the cloning and expression of NRPS TEs, the synthesis of thioester peptides, and the . biochemical characterization of the enzyme.

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Chemo-Enzymatic Synthesis of Non-ribosomal Macrolactams by a Penicillin-Binding Protein-Type Thioest their stand-alone nature, highlight PBP-type TEs as valuable candidates for development as biocatalysts for peptide macrocyclization. Here in this chapter, we describe the scheme for the chemoenzymatic synthesis of non-ribosomal macrolactam by SurE, a representative member of PBP-type TEs.

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Ribosomal Synthesis of Peptides Bearing Noncanonical Backbone Structures via Chemical Posttranslatiocursor residues and their chemical posttranslational modification processes. In this chapter, we describe the detailed procedures for the in vitro translation of peptides containing the precursor residues by means of genetic code reprogramming technology and posttranslational generation of objective noncanonical backbone structures.

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