Methods for producing ribosomally synthesized and posttranslationally modified peptides
The disclosure provides recombinant cells and methods for producing a ribosomally synthesized and posttranslationally modified peptide (RiPP), as well as RiPP libraries and methods for producing RiPP libraries.
1. A method for producing a ribosomally synthesized and posttranslationally modified peptide (RiPP), comprising combining a PatD or a TruD RiPP heterocyclase, a PatA or a TruA RiPP leader protease, a PatG or a TruG RiPP macrocyclase, RiPP heterocyclase substrate, and a sulfide source to form a mixture, under conditions sufficient to convert the RiPP heterocyclase substrate into a ribosomally synthesized and posttranslationally modified peptide;
wherein the RiPP is produced in vivo in a bacterial cell and/or in vitro.
2. The method of claim 1 , wherein the RiPP heterocyclase substrate is a TruE variant.
3. The method of claim 1 , wherein the RiPP heterocyclase substrate is a PatE variant.
4. The method of claim 1 , wherein the mixture further comprises a RiPP prenyl transferase.
5. The method of claim 1 , wherein E. coli produces the RiPP.
6. The method of claim 1 , wherein the RiPP is produced in vivo wherein the RiPP heterocyclase, the RiPP leader protease, and the RiPP macrocyclase are from an exogenous source.