• KSBB Journal
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• v.24 no.6
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• pp.563-569
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• 2009

Fibroin is an insoluble structural protein from Bombyx mori. It can be solubilized by dissolving in a hot $CaCl_2$ solution and subsequent dialysis. The aqueous solution is unstable and a transition from aqueous fibroin molecules rich in random coil is undergo to one rich in $\beta$-sheet content, resulting in hydrogelation. However, fibroin gel is so fragile and plastic that its mechanical property should be reformed for various applications. In this report, a semi-solid form of fibroin gel was prepared using glycerol and ethanol and was investigated to analyze their flow properties. A fibroin gel with 80% glycerol showed pseudoplastic and thixotropic properties. The square root of its yield stress varied linearly with fibroin concentration and it extrapolated to zero shear stress at 0.2% fibroin. A fibroin gel with 40% ethanol was shown to be highly thixotropic but its shear-thinning behavior was only observed above a certain level of shear rate. Its pseudoplasticity was restored by a high rate of shear stress.

• Journal of Sericultural and Entomological Science
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• v.41 no.1
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• pp.41-47
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• 1999

Silk fibroin dissolved in highly concentrated calcium chloride and ethanol mixture aqueous solution turned into gel under suitable conditions. Preparation conditions and properties of gel were investigated as a function of parameters such as pH of solution, fibroin concentration, glycerol concentration and molecular weight. When pH of silk fibroin aqueous solution was near the isoelectronic point(pH 3.9~4.0), gelation occurred rapidly and strength of gel was stonger than that of pH-unadjusted due to electrostatic repulsion decrease between silk fibroin macromolecules. As concentration of silk fibroin and glycerol was higher, gelation occurred more rapid. FT Infra-red spectra of freeze-dried fibroin gel showed that gelation was derived by intermolecular anti-parallel ${\beta}$-sheet structure formation. In addition to, it was found that white-precipitate occurred instead of gelation when aqueous silk fibroin was treated by enzyme(flavouzyme), however, after flavouzyme-treated silk fibroin aqueous solution was centrifugated gelation occurred instantly. The results of differential scanning thermal analysis and infra-red spectroscopy showed that thermal stability and crystallinity of enzyme-hydrolyzed fibroin are superior to those of unhydrolyzed fibroin.

• Proceedings of the Korean Society of Sericultural Science Conference
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• 2003.04a
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• pp.56-56
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• 2003

Using gel filtration chromatography (GFC), pure separation of high molecular silk fibroin was obtained and silk fibroin microsphere particles (SFMP) could be simply made by spray dryer method. Also, some of its physicochemical properties and morphology were investigated. The average molecular weight (Mw) of pure silk fibroin protein dissolved in calcium chloride is about 61, 500 g/mol as measured by gel permeation chromatography(GPC). (omitted)

• International Journal of Industrial Entomology and Biomaterials
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• v.2 no.1
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• pp.7-13
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• 2001

Enzymatic hydrolysis of different forms of silk fibroin (soluble, gel and insoluble forms) by industrial and commercial enzyme preparations to obtain aqueous and powdered silk fibroin in relatively mild conditions was investigated. A mono-enzymatic hydrolysate systems were tested for hydrolysis of water-soluble form of fibroin as most productive form of protein substrate. Insoluble forms of substrate usually were hydrolyzed less effective. In some cases from soluble fibroin substrate gel was formed during hydrolysis process. This hindered intermixing and decreased rates of hydrolysis. Insoluble sediments were formed in enzymatic hydrolysates in other cases. These sediments and also sediment after chemical hydrolysis were purified and tested on amino acids content for comparison. Sediments formation in these conditions are considered as pure tyrosine isolation method. Obtained hydrolysates were characterized by gel-chromatography analysis and other standard biochemical methods. Possibility of application of enzymatic hydrolysis for preparation of silk fibroin hydrolysates is discussed.

• International Journal of Industrial Entomology and Biomaterials
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• v.25 no.1
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• pp.75-79
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• 2012

Low-melting-temperature agarose gel solution, as a novel porogen was combined with a silk fibroin solution to generate interconnected porous networks. The porosity of the resultant silk fibroin-agarose scaffolds was greater than that of the scaffolds generated with agarose and deionized water. The porosities of silk fibroin scaffolds containing agarose gel at 0.5%, 1.0%, 1.5%, 2.0% [w/v] were 110.9%, 111.7%, 120.9%, and 123.0%, respectively. Lastly, the internal space generated in scaffolds after dissolution of the agarose gel provides a good environment for cell growth and movement within the scaffold.

• Journal of Sericultural and Entomological Science
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• v.41 no.3
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• pp.211-215
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• 1999

The pure-separation of calcium chloride-treated fibroin hydrolysates could be carried out using gel filtration chromatography. Also, the effect of its enzymatic hydrolysis was investigated in order to find out the enhancement of their functionality. The average molecular weight(Mw), solubility and free amino acid compositions of three hydrolysates samples (calcium chloride, calcium chloride-flavourzyme and calcium chloride-thermoase)were measured to compare their characteristics. The molecular weight of calcium chloride hydrolysate was about Mw 46,800 and it can be reduced to Mw 12,500 and 1,070 upon the enzymatic hydrolysis by flavourzyme and thermoase, repectively. A solubility of calcium chloride-treated samples shows about 60% while calcium chloride/enzyme-treated samples are perfectly soluble (100% solubility). The total amino acid composition of calcium chloride enzymatic hydrolysates are much higher than that of calcium chloride hydrolysate.

• Korean Journal of Food Science and Technology
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• v.40 no.4
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• pp.470-473
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• 2008

This study examined the enzymatic digestion, average molecular weight distribution and structural changes of Bombyx mori silk gland fibroin using gel penneation chromatography and nuclear magnetic resonance methods. The pure-separation of calcium chloride-treated fibroin hydrolysates was carried out by gel filtration chromatography. Also, the effects of fibroin's enzymatic hydrolysis were investigated using an edible enzyme. The average molecular weight of three hydrolysate samples (silk gland fibroin (SF), SF-calcium chloride (SFC), and SFC-enzyme) were measured to compare their characteristics. The molecular weights of SF, SFC, and SFC-enzyme were approximately 108,000, 65,000, and 1,000 Da, respectively. Finally, we determined the structural characteristic changes of the different enzymatically digested samples by $^{13}C$ nuclear magnetic resonance methods. For the enzymatically digested fibroin, the glycine $^{13}C^{\alpha}$ resonance indicated that the amino acid was dramatically changed and/or separated out; however, this was not shown for the normal Bombyx mori silk gland fibroin.

• International Journal of Industrial Entomology and Biomaterials
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• v.6 no.2
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• pp.151-155
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• 2003

Using gel filtration chromatography, high molecular silk fibroin with high purity was obtained and silk flbroin microsphere particles (SFMP) could be simply made by spray dryer method. Also, some of the physicochemical properties of SFMP and morphology were investigated. The average molecular weight of pure silk fibroin protein dissolved in calcium chloride is about 61,500g/㏖ as measured by gel permeation chromatography. SFMP was spherical in shape, and particles, sized average of 2 ${\pm}$ 10 ${\mu}$, were observed by SEM and particle analyzer, respectively. Obtaining microspheres particles by spray dryer method accelerated the transition from the random coil to the $\beta$-sheet structure during spray dryer treatment. It was identified by the basic fourier transform infrared spectroscopy of SFMP. The swelling ratio of SFMP is majorly dependent on the pH of the solution, not on the occurred gelation. The characteristic structure, which might be applicable to immobilization of drugs is superior to other matrix materials for the use of biomaterials with skin affinity.

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.621-625
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• 2003

In order to investigate the properties of aqueous fibroin solution, the molecular weight distribution of silk fibroin was determined by gel filtration chromatography. The molecular weights spreaded from 200 kDa to less than 20 kDa. The distribution of molecular weight was significantly affected when the pH of solubilization solution is less than 1. Distributions of fibroin solution stored at various condition were also investigated.

• Journal of Sericultural and Entomological Science
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• v.43 no.1
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• pp.41-48
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• 2001

The purification, dissolution and powdering of stained waste silk obtained from weaving and dyeing process were studied for the surface modification of textile fabric and plastic materials. The whiteness of stained waste silk could be improved through degumming and bleaching with sodium hydrosulfite. The water-soluble fibroin solution can be obtained by dissoving the degummed waste silk in a boiling solution of 50% calcium chloride for 60 minutes. The salts and heavy metals contained in fibroin solution were removed by electric dialysis, wool fiber filtration and gel filtration chromatography. The fibroin powder was prepared by using a fine grinder after the alkali treatment for weakening the silk fiber. The fine fibroin powder of particle size around 30 ㎛ was obtained with a ultra fine-mill, while it was finer below 10 ㎛ with a ball-mill. The dissolved or powdered silk was applied to the surface of fabric with addition of the binder (a urethane resin). The moisture content of polyester and nylon fabrics treated with the silk solution was improved due to hygroscopic property of silk. The fine fibroin powder mixed with the binder ws coated on the surface of synthetic film by use of the air pressed sprayer. It was revealed that the hygroscopicity as well as the softness of fibroin powder coated film was much improved. Therefore, it is thought that the fine silk fibroin powder is applicable as an coating agent for the surface modification of plastic and synthetic leather.