Preparation of undegraded native molecular fibroin solution from silkworm cocoons

Yamada, Hiromi; Nakao, Hiroshi; Takasu, Yoko; Tsubouchi, Kozo

doi:10.1016/s0928-4931(01)00207-7

Cited by 277 publications

(293 citation statements)

References 5 publications

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“…This was followed by washing several times with deionized water and the degummed silk was left drying at room temperature. To obtain aqueous SF solution, 1.2 g degummed silk was added to 20 times (v/w) of Ajisawa's reagent (CaCl 2 /ethanol/water) (Yamada et al, 2001). The mixture was stirred at 78 • C to form a clear solution for 2 h. The resulting SF solution was then dialyzed against deionized water for at least 3 days at sub-ambient temperature to remove the neutral salts using cellulose tubing.…”

Section: Methodsmentioning

confidence: 99%

pH- and electro-responsive characteristics of silk fibroin–hyaluronic acid polyelectrolyte complex membranes

Malay

Batıgün

Bayraktar

2009

International Journal of Pharmaceutics

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a b s t r a c t pH-responsiveness of recently developed silk fibroin (SF) and hyaluronic acid (HA) polyelectrolyte complex (PEC) membranes and their potential use in electro-responsive drug release systems were investigated. PEC membranes were prepared within a narrow pH window (3.0-3.5) for a SF-HA weight ratio of 20 and they were characterized by Atomic Force Microscopy in addition to characterization studies previously reported by our group. Swelling kinetics of the membranes was studied for a pH window of 2.5-7.4 and cyclic swelling test was performed to determine the pH-responsiveness of the membranes. It was shown that membranes swelled more in alkaline conditions and responded to variations in pH of the medium. Electric-stimuli assisted drug permeation and release studies were performed with a custom-made diffusion cell under both passive condition and electric field applied in pulsatile fashion. The instantaneous flux raised as the current was applied and then declined when the current application was terminated, and this process was repeated on subsequent applications. SF-HA complex membranes were found promising for the electric-stimuli-sensitive release of a high molecular weight and charged model drug for a membrane-permeation controlled formulation.

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Section: Methodsmentioning

confidence: 99%

pH- and electro-responsive characteristics of silk fibroin–hyaluronic acid polyelectrolyte complex membranes

Malay

Batıgün

Bayraktar

2009

International Journal of Pharmaceutics

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“…This corresponds to a specific productivity of about 80 ug protein/cell/day. By comparison, the best recombinant high-yield mammalian cell culture systems only produce about 50 pg protein/cell/day (Altman et al, 2003;Yamada et al, 2001).…”

Section: Plasmid Dna Constructionmentioning

confidence: 99%

Expression of the cyan fluorescent protein in fibroin H-chain of transgenic silkworm

Goo¹,

Choi²,

Kim³

et al. 2017

International Journal of Industrial Entomology

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We constructed the fibroin H-chain expression system to produce enhanced cyan fluorescent proteins (ECFP) in transgenic silkworm cocoon. Fluorescent cocoon could be made by fusing ECFP cDNA to the heavy chain gene and injecting it into a silkworm. The ECFP fusion protein, each with N-and C-terminal sequences of the fibroin H-chain, was designed to be secreted into the lumen of the posterior silk glands. The expression of the ECFP/H-chain fusion gene was regulated by the fibroin H-chain promoter. The use of the 3xP3-driven EGFP cDNA as a marker allowed us to rapidly distinguish transgenic silkworms. The EGFP fluorescence became visible in the ocelli and in the central and peripheral nervous system on the seventh day of embryonic development. A mixture of the donor and helper vector was micro-injected into 1,020 Kumokjam, bivoltin silkworm eggs. We obtained 6 broods. The cocoon was displayed strong blue fluorescence, proving that the fusion protein was present in the cocoon. Accordingly, we suggest that the ECFP fluorescence silk will enable the production of novel biomaterial based on the transgenic silk.

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“…In the literature, the degradation was evaluated by the changes of silk mechanical properties [34], colour [30], crystalline structure by X-ray diffraction [14,18,35], viscosity [36,37], molecular weight of silk component by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) [38] and molecular weight distribution by SEC [19,30,36,[39][40][41]. As the macroscopic properties (like mechanical endurance) of silk textiles may be reflected by the distribution of fibroin chain length, condition of the historical silk textiles can be evaluated from measurements of the molecular weight distribution of the silk [ref].…”

Section: Introductionmentioning

confidence: 99%

Evaluating the impact of different exogenous factors on silk textiles deterioration with use of size exclusion chromatography

et al. 2016

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Size exclusion chromatography (SEC), especially coupled with multiple angle laser light scattering detector (MALLS) is a powerful tool in diagnostics of deterioration of historic and art objects to evaluate their condition. In this paper, SEC-UV-MALLS-DRI technique was applied to study degradation of silk fibroin samples (Bombyx mori) artificially aged under various conditions: in the presence of oxygen, in different amount of water vapour and in volatile organic products (VOCs), all at temperature of 90°C. Conditions were chosen in such a way that it mimicked real conditions of textiles' storing during exhibitions and in show cases. The influence of temperature, moisture and VOCs content on the state of silk textiles was examined with the use of size exclusion chromatography. Pseudo-zero-order Ekenstam equation was applied to study degradation rates of fibroin with use of the approximated values of DP of fibroin.

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Preparation of undegraded native molecular fibroin solution from silkworm cocoons

Cited by 277 publications

References 5 publications

pH- and electro-responsive characteristics of silk fibroin–hyaluronic acid polyelectrolyte complex membranes

pH- and electro-responsive characteristics of silk fibroin–hyaluronic acid polyelectrolyte complex membranes

Expression of the cyan fluorescent protein in fibroin H-chain of transgenic silkworm

Evaluating the impact of different exogenous factors on silk textiles deterioration with use of size exclusion chromatography

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