Recombinant spidroin production and fabrication using crude-enzyme-treated corn stalk medium

Corn stalk residues represent a potential feedstock for recombinant protein materials. This study integrated corn stalk treatment using crude cellulase and laccase with recombinant spidroin production and film fabrication. Following autoclaving and enzymatic treatment at 65 °C for 8 h, the combined treatment achieved a DNS-measured reducing sugar concentration of 141.2 ± 1.6 g/L, compared with 56.3 ± 4.9 g/L in the no-enzyme control, accompanied by approximately 47% solid mass loss and surface disruption. The resulting medium supported Escherichia coli growth and spidroin expression, enabling fabrication of transparent, flexible films. Representative films exhibited maximum tensile stresses up to 40.4 with estimated β-sheet fractions of 24.5%. These findings demonstrate the feasibility of using crude-enzyme-treated corn stalk medium for recombinant spidroin production and film fabrication, providing a basis for further development of agricultural residue valorization into protein-based materials.