Proteomic Level Solubilisation of Spider Silk by Two Step Heating strategy in an Aqueous Solution

Abstract

Spider silk is a proteinaceous fiber with remarkable mechanical properties spun from spider silk

proteins (spidroins).

Spider silk is a protein-based fiber with outstanding mechanical properties, composed primarily of spider silk proteins (spidroins). In nature, spidroins are spun directly into fibers, whereas under controlled in vitro conditions they can be processed into a variety of morphologies, such as films, hydrogels, fibers, capsules, and particles. The structural versatility and biochemical properties of silk proteins make silk-derived materials promising candidates for applications in tissue engineering, regenerative medicine, and the controlled delivery of protein drugs and peptide vaccines. In this work, we present a two-step heating solubilization strategy to extract spidroins from the web of Neoscona mukerjii, taking advantage of their high thermal stability in concentrated urea solutions. The influence of pH and temperature on the solubilization efficiency was examined, and the optimized conditions were determined as follows: heating at 55 °C-90 °C for upto 60 minutes, pH 9.0–10, and 7-8 M urea in standard biological buffers. The spidroins obtained through this method exhibited the capacity to self-assemble into spherical nanoparticles with smooth morphology. Compared with conventional solubilization approaches, this two-step heating method provides a more effective strategy for dissolving spider silk proteins, which may be valuable for downstream applications. Nevertheless, care should be taken in cases where urea-induced modifications could interfere with the intended application. Furthermore, the method demonstrates adaptability across a wide range of buffer systems, pH levels, and temperature conditions, suggesting its potential applicability for solubilizing other thermally stable proteins.