Engineered biological systems can enable the template-directed synthesis of biopolymers such as proteins and peptides with desirable material properties and functions. The cellular engine of this process, the ribosome and associated translation apparatus, drives the polymerization of hundreds to thousands of amino acid monomers into sequence-defined biopolymers with speed, efficiency, and accuracy unmatched by chemical methods. This extraordinary biosynthetic capability has been leveraged through recombinant protein production to address a broad range of challenges in the medical and industrial biotechnology industries. In nature, however, template-directed production of biopolymers is constrained to the 20 canonical amino acids, with limited ability to encode and incorporate unnatural monomers. The potential for harnessing the translation apparatus to manufacture new biopolymer-based products beyond these natural limits remains underexploited, and fundamental constraints on the efficiency and scalability of existing systems remain a challenge. Pearl Bio is uniquely positioned to overcome this barrier, with a team and technology to enable the production of new classes of sequence defined biopolymers with properties unmatched by biology or chemistry alone. The proposed work seeks to expand natures repertoire of ribosomal monomers to non-canonical amino acids (ncAAs) in a scalable production system that can yield new classes of materials, chemicals, and enzymes with diverse genetically encoded chemistries. Pearl Bio is collaborating with research institution partner Northwestern University, establishing a team with deep experience (i) expanding the scope of ribosome-mediated polymerization to include chemically diverse scaffolds, (ii) engineering and evolving the translation apparatus, (iii) constructing genomically recoded organisms (GROs) with new genetic codes and open codons that can be dedicated to ncAA incorporation, and (iv) producing novel biomaterials harboring multiple ncAAs. This experience will be leveraged in Phase I to establish feasibility for incorporation of two distinct ncAAs in a biopolymer using a novel GRO, enabling production of sequence-defined molecules that have never been synthesized before. Both cell-based and cell-free production systems will be developed, enabling versatile approaches to accelerate achievement of manufacturing-relevant titers of prototype designer proteins in Phase II.
