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 culturing a recombinant cell in the presence of exogenous mevalonate, wherein the recombinant cell comprises a nucleic acid encoding a tru protein or a pat protein, and wherein the recombinant cell comprises a nucleic acid encoding a mev protein.
2. The method of claim 1 , wherein the recombinant cell expresses ERG12, ERG8, and MVD1.
3. The method of claim 1 , further comprising culturing the recombinant cell in the presence of an exogenous sulfide source.
4. The method of claim 3 , wherein the exogenous sulfide source comprises cysteine.
5. The method of claim 4 , wherein cysteine is provided at a concentration of 0.5-40 mM.
6. The method of claim 4 , wherein cysteine is provided at a concentration of 5-10 mM.
7. The method of claim 3 , wherein the exogenous sulfide source comprises cystine.
8. The method of claim 3 , wherein the exogenous sulfide source comprises hydrogen sulfide.
9. The method of claim 3 , wherein the exogenous sulfide source comprises sodium hydrosulfide.
10. The method of claim 1 , wherein the recombinant cell expresses a TruE variant.
11. The method of claim 1 , wherein the recombinant cell expresses a PatE variant.
12. The method of claim 1 , wherein the exogenous mevalonate is provided at a concentration of 5-40 mM.
13. The method of claim 1 , wherein the recombinant cell is Escherichia coli.