Orthogonal Translation-Based Strategy for Growth-Decoupled Protein Production

Abstract

Optimizing resource allocation between growth and production remains a major challenge in microbial protein manufacturing. In many systems, excessive biomass accumulation limits the efficient synthesis of recombinant proteins, leading to reduced yield and productivity. To address this limitation, we developed a modular genetic system that enables phase-specific control of protein synthesis in a two-stage bioprocess. By separating growth and production phases, the system reduces cellular burden during early culture and enhances target expression when production is triggered. Through the use of orthogonal translation machinery and tunable regulatory elements, we dynamically modulated translational activity and improved protein yield Application of this approach led to improved protein yields and reduced metabolic stress in the host strain. The framework is versatile and can be extended to various target proteins and microbial production systems. This strategy provides a generalizable solution for growth-decoupled protein production, contributing to more efficient and scalable microbial biomanufacturing.