Spider silk fibers are stronger and tougher than industrial fibers while being biodegradable and sustainable. Artificial spider silk products exist, but their rational design and cost-effective production at scale remains challenging. Spider silk-like proteins, essential for these fibers, can be made using microbes like E. coli, but yields are low compared to other proteins. The fungus T. reesei could scale up production, but its enzymes degrade silk proteins due to their disordered nature. On the other hand, standard methods can’t fully characterize flexible and disordered regions of silk proteins. This disordered nature gives spider silk its unique properties but creates challenges in production and design. This project tackles these challenges using computational and experimental methods. The goals are to understand how protein sequences and processing conditions can be rationally optimized, and provide tools to improve production by increasing solubility and reducing degradation.