13C NMR study of silk fibroin synthesis in vitro
AbstractAbstract
[en] The posterior silkgland of Bombyx mori, silkworm larvae, produces abundant specific silk fibroin at about 80 μg/cell for one day. In this study, the continuous production of silk fibroin in the posterior silk-gland was monitored by circumfusion cultivation using both C-13 NMR and C-13 labeling techniques in order to quantitatively determined the production of silk fibroin in vitro. The experimental setup is shown. The posterior silkglands of four larvae were collected in a 10 mm NMR sample tube under sterile condition. The C-13 NMR spectra were observed in the culture medium. The time-dependence of the C-13 NMR spectra of the culture medium containing the posterior silkglands are shown. The peak intensity for silk fibroin increased with the lapse of time. Also the peak due to the Cα carbon of the serine residue of silk fibroin slightly increased, which indicates that there is a system which converts glycine to serine in the posterior silkglands. In order to discuss this silk production quantitatively, the relative intensity of the Cα peaks for silk fibroin and glycine in the cultivation medium was plotted. The ability of the posterior silk-glands to produce silk protein was still high in vitro when the circumfusion cultivation technique was used. (K.I.)
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