Chicken feather, a potential source of keratin, is often disposed as waste material. Although some methods, i.e., hydrolysis, reduction, and oxidation, have been developed to isolate keratin for composites, it has been limited due to the rising environmental concerns. In this work, a green solvent N-methylmorpholine N-oxide (NMMO) was used to extract keratin from chicken feather waste. Eighty-nine percent of keratin was extracted using 75% NMMO solution. However, the result from size exclusion HPLC showed that most of the keratin degraded into polypeptide with molecular weight of 2189 and only 25.3% regenerated keratin was obtained with molecular weight of 14,485. Analysis of amino acid composition showed a severe damage to the disulfide bonds in keratin during the extraction procedure. Oxidization had an important effect on the reconstitution of the disulfide bonds, which formed a stable three-dimensional net structure in the regenerated keratins. Besides, Raman spectra, NMR, FT-IR, XRD, and TGA were used to characterize the properties of regenerated keratin and raw chicken feather. In the end, a possible mechanism was proposed based on the results.