Engineering ribosomally synthesized and post-translationally modified peptide biosynthetic pathways for de novo peptides with designed structures and programmed functions

Engineering of ribosomally synthesized and post-translationally modified peptide (RiPP) biosynthetic pathways has emerged as a powerful strategy for generating peptides with designed structures and programmed functions. RiPP biosynthetic machineries combine genetic programmability with the broad substrate tolerance of post-translational modification enzymes, allowing systematic diversification of peptide structures. Recent advances in RiPP engineering have enabled diverse strategies for constructing peptides with expanded chemical complexity and tailored biological activities. This review describes two major directions in current RiPP engineering. One focuses on the integration of biosynthetic machineries from distinct RiPP classes to construct hybrid RiPPs, in which multiple class-specific non-proteinogenic building blocks coexist within a single peptide scaffold. The other exploits rational sequence engineering and genetically encoded peptide library technologies to develop de novo RiPP derivatives with programmed biological functions. These complementary strategies highlight the utility of RiPP biosynthetic pathway engineering for exploring peptide chemical space and creating functional molecules beyond natural biosynthesis.