Bombyx mori silk fibroin (BMSF) has received considerable research interest as a potential biomaterial owing to its excellent mechanical properties and benign, versatile material fabrication options, including electrospinning. Despite this, characterizations of regenerated BMSF aqueous solutions and electrospun materials resulting from them are still very limited in the literature. This report details the rheological characterization of regenerated aqueous BMSF solutions under shear and elongational deformation. Well-characterized regenerated BMSF solutions were then systematically electrospun over a range of concentrations and process parameters to determine their effects on electrospinning processing windows and fiber morphology. BMSF solutions could not be electrospun successfully if BMSF concentration was below 20 wt % or the relaxation time measured using the CaBER rheometer was below 0.001 s. Electrospun BMSF fiber diameter was found to increase with solution concentration when stable electrospinning was achieved. An upper threshold of 30 wt % BMSF solution was identified for the formation of fibers with a circular cross section. Adding small amount of high molecular weight poly(ethylene oxide) was an effective rheological modifier that greatly improved the electrospinnability of BMSF solutions. Electrospinning BMSF-PEO solutions over a range of parameters significantly altered the fiber products. Increasing voltage from 0.5 to 1 kV/cm was found to decrease fiber diameter by approximately 50% (p < 0.001). Flow rate was found to have a significant effect on fiber diameter, which decreased with spinneret height. The results presented here provide valuable guidance in the production of BMSF electrospun materials with specific properties for tissue engineering and regenerative medicine.