화학생물학 연구실

High-performance cell-free protein synthesis has transformative potential for synthetic biology, yet the prohibitive costs of PURE kits and the labor intensity of in-house preparation have restricted accessibility and scalability. We developed i-POPFLEX (Purified components OPtimized for FLEXible protein expression), a modular cell-free protein synthesis system in which 34 translation factors and a split T7 RNA polymerase are individually synthesized in vitro and assembled using automated liquid handling. This workflow minimizes manual input and supports parallelized production, generating complete, ready-to-use systems within two days. i-POPFLEX achieves up to 8.4-fold higher protein yields and a 96 % cost reduction (27-fold lower cost) compared with commercial kits. Its flexible architecture also enables selective component inclusion for genetic code reprogramming and site-specific incorporation of non-canonical amino acids. By coupling modular design with automation, i-POPFLEX provides an accessible, customizable, and economically viable platform for next-generation biomanufacturing workflows. Cyclic motifs, found in diverse natural products such as penicillin V, goadsporin, and patellamide C, are widely recognized for their exceptional pharmacological efficacy. Traditionally, these cyclic structures have been produced through indirect methods, including enzymatic post-translational modifications, where a linear peptide is first synthesized by the ribosome and then cyclized by specific enzymes or solid-phase peptide synthesis (SPPS). However, these approaches often face limitations in terms of productivity, overall efficiency, and the scalability required for constructing diverse, high-throughput libraries. To address these challenges, our research focuses on the rib