Engineered membraneless organelles in for enhanced indigoidine biosynthesis and antimicrobial peptide production.

Liquid-liquid phase separation (LLPS)-driven membraneless organelles (MLOs) have been employed to enhance metabolic efficiency in various microbial cell factories. However, their application in the industrial bacterium has not been explored. Here, we report the formation of liquid protein condensates in using the RGG domain of LAF-1. We optimized conditions for condensate formation, including the pre-induction period, inducer concentration, and cultivation temperature. Using the indigoidine biosynthesis pathway as a model, we demonstrated that LLPS-mediated MLOs enhanced indigoidine production. Furthermore, we applied these MLOs to modulate the toxicity of antimicrobial peptides (AMPs) to host cells, facilitating the expression of AMPs, including melittin and lactoferricin B. These findings provide insights into MLOs engineering in and suggest broader applications of LLPS-mediated systems in industrial biotechnology.