Silk
fibroin is a protein with a unique combination of properties
and is widely studied for biomedical applications. The extraction
of fibroin (degumming) from the silk filament impacts the properties
of the outcoming material. The degumming can be conducted with different
procedures. Among them, the most used and studied procedure in the
research field is the alkali degumming with sodium carbonate (Na2CO3). In this study, by the use of a statistical
method, namely, design of experiment (DOE), we characterized the Na2CO3 degumming, taking into consideration the main
process factors involved and changing them within a selected range
of values. We considered the process temperature and time, the salt
concentration, and the number of baths used, testing the impact of
these variables on the fibroin properties by building empirical models.
These models not only took into consideration the direct effect of
the process factors but also their combined effect, which are not
conventionally detectable with other methods. The weight loss and
the amount of sericin removed in the process were determined and used
as a measure of the effectiveness of the process. The secondary structure,
the molecular weight, the diameter of fibers, and their morphology
and mechanical properties were studied with the intent to correlate
the macroscopical properties with the structural changes. We report,
for the first time, the possibility to effectively remove all sericin
from the silk fibroin using Na2CO3, using a
process that requires less salt, water, and energy, in comparison
with the standard alkali protocol, making this technique overall more
environmentally sustainable; in addition, we have demonstrated the
possibility to tune the material properties by varying the degumming
conditions and even to optimize them with empirical statistically
based equations that allow one to directly set the optimal process
parameters. The major effect on the macroscopical properties (such
as the ultimate strength and Young’s modulus) has been proved
to be correlated with the removal of sericin instead of the microstructural
variations. Finally, a ready-to-use table with a set of optimized
degumming procedures to maximize or minimize the studied properties
was provided.