A facile and eco-friendly approach for preparation of microkeratin and nanokeratin by ultrasound-assisted enzymatic hydrolysis

Protein is one of the most abundant natural polymeric materials, but only a few studies on nanoproteins have been conducted. In this paper, a novel approach based on ultrasound-assisted enzymatic hydrolysis was employed for the preparation of microkeratin and nanokeratin from wool. The hydrolysis system included a solution containing enzyme (esperase) and reductant (L-cysteine) and treated ultrasonically to remove the scales and amorphous regions within wool. Results showed that the reaction was most effective at 50 °C and pH 7, when incubated for 3 h, followed by sonication for 6 h. The products included spindle-shaped microkeratin (4–7 μm in diameter and 70–120 μm in length) and cone-shaped nanokeratin (50–300 nm in diameter and less than 15 μm in length). Under ultrasonic-assisted conditions, the yields of microkeratin and nanokeratin increased significantly, while the treatment time decreased. Fourier transform infrared spectroscopy (FTIR) showed that the chemical structures of microkeratin and nanokeratin did not change, compared to that of wool. X-ray diffraction (XRD) analysis showed that the microkeratin was mainly composed of α-helical structure, while the β-sheet structure was more prevalent in nanokeratin. The presented method is facile and eco-friendly, thereby paving new pathways for the preparation of microkeratin and nanokeratin.