One-pot biosynthesis of genetically encoded amphiphiles via protein prenylation

Despite the well-established role of post-translational lipidation in regulating diverse facets of cellular biology, scalable access to lipidated proteins remains limited by reliance on complex eukaryotic expression systems or labor-intensive semisynthetic approaches. To address these limitations, we describe a robust, genetically encoded platform for production of prenylated proteins in prokaryotes through metabolic and genetic engineering. We apply this system to synthesize prenylated intrinsically disordered protein polymers derived from the elastin consensus sequence as genetically encodable amphiphiles as a model system to investigate how prenylation syntax controls self-assembly and phase behavior. We further demonstrate the platform's versatility by extending it to natively geranylgeranylated globular proteins, such as RhoA, establishing a scalable, protein-agnostic toolkit for biomaterials and biochemical research.