We have recorded several kinds of
13C NMR to characterize the structure of the noncrystalline domain in
Bombyx mori silk fibroin and the influence of hydration on the structure of silk fibroin. First, the structural analysis of the soluble fraction (noncrystalline domain) after chymotrypsin hydrolysis of the silk fibroin has been performed by
13C solution NMR spectroscopy. In the noncrystalline fraction. Cs fraction, the
C-terminal amino acid residues are identified as---Gly-Tyr from the pH titration of the spectra. Similarly, the
N-terminal residues are identified as Gly-Ser---. Second, the
13C CP/MAS NMR spectra for the precipitated fraction (Cp fraction; crystalline domain) and Cs fraction have been observed as a function of moisture regain in order to know what parts, residue or groups, interact with the adsorbed water. In the Cp fraction, the significant spectral changes were not observed with increasing water content. However, in the Cs fraction, the significant changes were observed at the carbonyl carbon peak of the
N-terminal Gly residue and the aromatic C
γ carbon peak of the
C-terminal Tyr residue by hydration. Third, the
1H NMR spin-lattice relaxation times, T
1 of water in Cp and Cs fractions were observed. The T
1 values were analyzed as two components for the Cp fraction and three components for the Cs fraction. The water molecules with the restricted motion increase considerably in the Cs fraction and such a restricted motion is roughly divided into two cases. This seems to be corresponding to the strongly hydration at the terminal groups observed in the
13C CP/MAS NMR spectrum of the Cs fraction.
View full abstract