• Journal of Sericultural and Entomological Science
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• v.45 no.1
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• pp.46-57
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• 2003

This study was carried out to investigate the characteristics of the soluble sericins after degumming and after hydrolysis of insoluble sericin with various enzymes. Especially, the hydrolysis characteristics were examined in terms of molecular weight of the soluble sericin. Amino acid composition and molecular weight characteristics of the soluble sericins were also studied. When the insoluble sericin was hydrolyzed with kojizyme and flavourzyme, the solubility was highest at pH 7 and 50$^{\circ}C$. On the other hand, in the cases of protamex and alcalase, the highest solubility was obtained at 60$^{\circ}C$. In these cases, solubility increased with pH. In enzymatic hydrolysis, the solubility was increased with concentration of enzymes until 4 hours. After then, a slight difference was found along with treatment times. In enzymatic hydrolysis, the absorbance of the soluble sericin was increased with concentration of enzymes and treatment times. Average degree of polymerization was decreased with treatment time and concentration. The amino acid compositions were similar in low(low molecular weight by degumming) and high (high molecular weight by degumming). Those of PK (soluble sericin hydrolyzed with kojizyme), PP (soluble sericin hydrolyzed with protamex), and PA(soluble sericin hydrolyzed with alcalase) were similar to each other. Serine and tyrosine compositions were higher in low and high than those of PK, PP, and PA. However proline was absent in low and high. Molecular weights of the various sericins became higher as KP>high>PP>low>PA and those of KP and PA were 9,800 and 905 respectively.

• Journal of Sericultural and Entomological Science
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• v.45 no.1
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• pp.58-65
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• 2003

This study was carried out to investigate the characteristics of the soluble sericins after degumming and after hydrolysis of insoluble sericin with various enzymes. Physical and chemical characteristics of the soluble sericins were also studied and pack test was conducted. Electro donating ability of the various sericins became higher as high＞low＞KP＞PP＞PA. In FT-IR analysis, the band of insoluble sericin at 3,285cm$\^$-1/ was shifted to the lower wavelengths in soluble sericins. The intensity of the amide II band was reduced and shifted to lower wavelengths as a result of hydrolysis. In DSC analysis, all sericins showed endothermic peaks around 200$^{\circ}C$ and insoluble sericin showed another peak at 250$^{\circ}C$. The endothermic peak of the insoluble sericin was found at higher temperature compared with those of the soluble sericins which showed at 300$^{\circ}C$. Whiteness of the various sericins became higher as low＞high＞PP＞PA＞PK＞IN and yellowness were PK＞PA＞PP＞low＞high. In pack test, the skin packed with the non-woven fabric treated with the various sericins was more smooth and less oily.

• Journal of Sericultural and Entomological Science
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• v.42 no.2
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• pp.104-108
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• 2000

To hydrolyze insolule sericin the enzyme hydrolysis was used, and then obtained the results as given belows. When insoluble sericin was hydrolyzed by enzyme treatment, the solubility was best at pH 7, 60$\^{C}$ and was slightly increased both above 2 hours treatment and above 10% of enzyme concentration. As the results of electrophoresis, the distribution of molecular weight of sericin powder obtained by enzyme hydrolysis was very weak and showed in the wide range having no distinguishable band. Average degree of polymerzations (A.D.P.) of sericin hydrolyzed by enzyme were about 4.1∼6.3, average molecular weight were about 470∼730. The whiteness of the sericin powder obtained by enzyme hydrolysis was high and increased slightly with higher treatment concentration of enzyme. As the results of amino acid analysis, the amino acid analysis, the amino acid composition of the sericin powder from the enzyme treatment were similar to which located at near 230$\^{C}$ and 320$\^{C}$. The peak of near 230$\^{C}$ could not been found in the sericin powder obtained by enzyme hydrolysis.

• Journal of Sericultural and Entomological Science
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• v.16 no.1
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• pp.21-48
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• 1974

The studies were carried out to disclose the physical and chemical properties of sericin fraction obtained from silk cocoon shells and its characteristics of swelling and solubility. The following results were obtained. 1. The physical and chemical properties of sericin fraction. 1) In contrast to the easy water soluble sericin, the hard soluble sericin contains fewer amino acids include of polar side radical while the hard soluble amino acid sach as alanine and leucine were detected. 2) The easy soluble amino acids were found mainly on the outer part of the fibroin, but the hard soluble amino acids were located in the near parts to the fibroin. 3) The swelling and solubility of the sericin could be hardly assayed by the analysis of the amino acid composition, and could be considered to tee closely related to the compound of the sericin crystal and secondary structure. 4) The X-ray patterns of the cocoon filament were ring shape, but they disappeared by the degumming treatment. 5) The sericin of tussah silkworm (A. pernyi), showed stronger circular patterns in the meridian than the regular silkworm (Bombyx mori). 6) There was no pattern difference between Fraction A and B. 7) X-ray diffraction patterns of the Sericin 1, ll and 111 were similar except interference of 8.85A (side chain spacing). 8) The amino acids above 150 in molecular weight such as Cys. Tyr. Phe. His. and Arg. were not found quantitatively by the 60 minutes-hydrolysis (6N-HCI). 9) The X-ray Pattern of 4.6A had a tendency to disappear with hot-water, ether, and alcohol treatment. 10) The partial hydrolysis of sericin showed a cirucular interference (2A) on the meridian. 11) The sericin pellet after hydrolysis was considered to be peptides composed with specific amino acids. 12) The decomposing temperature of Sericin 111 was higher than that of Sericin I and II. 13) Thermogram of the inner portioned sericin of the cocoon shell had double endothermic peaks at 165$^{\circ}C$, and 245$^{\circ}C$, and its decomposing temperature was higher than that of other portioned sericin. 14) The infrared spectroscopic properties among sericin I, II, III and sericin extracted from each layer portion of the cocoon shell were similar. II. The characteristics of seriein swelling and solubility related with silk processing. 1) Fifteen minutes was required to dehydrate the free moisture of cocoon shells with centrifugal force controlled at 13${\times}$10$^4$ dyne/g at 3,000 R.P.M. B) It took 30 minutes for the sericin to show positive reaction with the Folin-Ciocaltue reagent at room temperature. 3) The measurable wave length of the visible radiation was 500-750m${\mu}$, and the highest absorbance was observed at the wave length of 650m${\mu}$. 4) The colorimetric analysis should be conducted at 650mu for low concentration (10$\mu\textrm{g}$/$m\ell$), and at 500m${\mu}$ for the higher concentration to obtain an exact analysis. 5) The absorbing curves of sericin and egg albumin at different wave lengths were similar, but the absorbance of the former was slightly higher than that of the latter. 6) The quantity of the sericin measured by the colorimetric analysis, turned out to be less than by the Kjeldahl method. 7) Both temperature and duration in the cocoon cooking process has much effect on the swelling and solubility of the cocoon shells, but the temperature was more influential than the duration of the treatment. 8) The factorial relation between the temperature and the duration of treatment of the cocoon cooking to check for siricin swelling and solubility showed that the treatment duration should be gradually increased to reach optimum swelling and solubility of sericin with low temperature(70$^{\circ}C$) . High temperature, however, showed more sharp increase. 9) The more increased temperature in the drying of fresh cocoons, the less the sericin swelling and solubility were obtained. 10) In a specific cooking duration, the heavier the cocoon shell is, the less the swelling and solubility were obtained. 11) It was considered that there are differences in swelling or solubility between the filaments of each cocoon layer. 12) Sericin swelling or solubility in the cocoon filament was decreased by the wax extraction.. 13) The ionic surface active agent accelerated the swelling and solubility of the sericin at the range of pH 6-7. 14) In the same conditions as above, the cation agent was absorbed into the sericin. 15) In case of the increase of Ca ang Mg in the reeling water, its pH value drifted toward the acidity. 16) A buffering action was observed between the sericin and the water hardness constituents in the reeling water. 17) The effect of calcium on the swelling and solubility of the sericin was more moderate than that of magnecium. 18) The solute of the water hardness constituents increased the electric conductivity in the reeling water.

• Journal of Sericultural and Entomological Science
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• v.42 no.1
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• pp.31-35
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• 2000

When insoluble sericin was hydrolyzed by treatment of NaOH solution, the solubility was increased with higher treatment temperature and longer treatment time. Whereas it was decreased in addition of NaHSO$_3$. As the results of electrophoresis in sericin powder obtained by the NaOH treatment, a distinguishable band was not confirmed. Average degree of polymerizations(A.D.P.) of sericin hydrolyzed by NaOH solutions were about 19.6∼22.1 and average molecular weight(M.W.) were about 2,200∼2,500. The longer hydrolysis time increased the whiteness of sericin powder. As the results of amino acid analysis, the contents of Thr., Tyr., and Ser. were decreased in NaOH hydrolysis as compared to HCl hydrolysis. In DSC analysis, thermal deformation and pyrolysis peak located at near 230$\^{C}$ and 320$\^{C}$, respectively.

• Journal of Sericultural and Entomological Science
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• v.45 no.1
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• pp.34-45
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• 2003

Silk was treated with calcium hydroxide for degumming at different treatment times, temperatures and Ca(OH)$_2$ concentration to optimize degumming conditions in this thesis. After degumming, soluble and insoluble sericin were seperated and then the soluble sericin was characterized by measuring the average degree of polymerization (D.P.), lysinoalanine (LAL) content, DSC, and by amino acid analysis. And degummed silk fibroin was characterized by measuring tenacity and SEM. Degumming loss was increased by increasing the treatment time and temperature until about 30 minutes. After then, a slight difference was found along with treatment times at the Ca(OH)$_2$ concentrations of 0.07% and 0.1% solutions. After degumming, insoluble sericin ratio on degumming solution was increased by increasing treatment temperature at Ca(OH)$_2$ 0.04% solution. At the concentration Ca(OH)$_2$ of 0.07%, a soluble ratio was almost 100% regardless of treatment time and temperature. At the beginning of treatment, insoluble ratio was high at Ca(OH)$_2$ 0.1% solution but it was decreased by increasing treatment time. At the Ca(OH)$_2$ concentration of 0.04%, D.P. of soluble sericin was maintained as a constant value of 10 at 100$^{\circ}C$ although treatment time was increased. However, at 80$^{\circ}C$ and 90$^{\circ}C$, it was hard to prepare a soluble sericin having a constant D.P. by increasing treatment time. At the Ca(OH)$_2$ concentration of 0.07%, D.P. was almost 10 irrespective of treatment temperature and time. Soluble sericins with high D.P. of 20∼30 were obtained at 0.1% and 100$^{\circ}C$. LAL was not detected in soluble sericin. As the results of amino acid analysis, it showed that Ca(OH)$_2$ degumming reduced the contents of hydroxy amino acids like Ser., Thr. and Tyr. In DSC analysis of soluble sericin, endothermic peak by thermal deformation and pyrolysis showed at 189$^{\circ}C$ and at 299$^{\circ}C$, respectively. The tenacities of degummed silk were 15∼30% lower than that of raw silk. And it was decreased with increasing treatment time. From the morphological study, the thickness of degummed silk fibroin became thinner by increasing degumming loss. The roughness of a silk fibroin surface was appeared as treatment concentration was increased.

• Textile Coloration and Finishing
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• v.14 no.4
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• pp.249-258
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• 2002

Natural silk is formed by two proteins : the crystalline fibroin (inside the silk thread) and amorphous sericin (as a tube outside the thread). The degumming process is used to eliminate the external sericin prior to dyeing ; generally it makes use of soaps at about pH 10. Sericin is the protein constituent that "gums"together the fibroin filaments of cocoon silk. It constitutes about 25% of the weight of the cocoon, is soluble in hot water and "gels" on cooling. The removal of sericin from raw silk, known as degumming, is a simple but important process usually employing hot dilute soap or alkaline solution and occasionally dilute acids or enzymic methods. During degumming, alkali is taken up by the sericin and the free acid from the soap is formed ; this may be deposited on the fiber, reducing the rate of degumming and protecting it from hydrolysis. Alkali is often added to maintain or restore the pH of the baths, but it is rarely used alone, since it leaves the silk rather harsh in handle. If complete sericin removal is required as for printing, sodium carbonate may be added. If the pH of the bath exceeds 11, the fibroin is attacked. Recently, According to the development of electrolysis, we can be obtained the electrolytic reduction water(above pH 11.5) and electrolytic oxidation water (below pH 3). The aim of this work was to study a degumming process using electrolytic water and a possibility of sericin recovery. The new degumming process used electrolytic water operates at $95^\circ{C}$ for 2hr. without any reagents. The wastewater of this process are formed by a solution of sericin in water. This conditions suggest the study of a possible recovery of this protein (sericin) which has an amino acid composition suitable for many used in cosmetics, textile finishing agents, animal feeding, etc. The degumming process using electrolytic water is available to reduce treatment costs and pollute and at the same time to recover sericin.

• Journal of Sericultural and Entomological Science
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• v.41 no.1
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• pp.48-53
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• 1999

The study was carried out to investigate the effects of hydrolysis conditions such as treatment times and concentrations on the solubility of insoluble sericin using the hydrochloric acid solution. When insoluble sericin was hydrolyzed by HCl solution, the solubility was increased with the higher treatment concentration. As the results of electrophoresis of sericin powder obtained by the HCl treatment, a distinguishable band was not confirmed. Average degree of polymerizations(A.D.P.) of sericin hydrolyzed by HCl solution were about 4.2~5.9 and average molecular weights(M.W.) were about 470~670. The longer hydrolysis time reduced the whiteness of sericin powder. As the results of amino acid analysis, the amino acid compositions of the sericin powder from HCl treatment were sililar to that of insoluble sericin, but Tyr. and Arg. were not detected in the powder obtained by HCl treatment. In DSC analysis, thermal deformation and pyrolysis peak located at near 220$^{\circ}C$ and 330$^{\circ}C$.

• Journal of Life Science
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• v.20 no.8
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• pp.1249-1253
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• 2010

Sericin is a type of high molecular weight water-soluble glycoprotein surrounding fibroin (silk protein) that has been used as a cell culture supplement and accelerates cell proliferation in various serum-free media. The purpose of this study was to investigate the enhancing effect of thyroglobulin (Tg) secretion by sericin in thyrocytes, FRTL-5 cells. While Tg-mRNA expression was not enhanced, a secreted form of Tg was obviously increased by sericin. In this status, expression of both endoplasmic reticulum (ER) molecular chaperones (Bip & calreticulin) and ER membrane proteins (IRE1, PERK & ATF6) was enhanced. The proximal step of IRE1, XBP1 mRNA splicing was slightly detected however, the proximal step of PERK, phosphorylation of $eIF2{\alpha}$, was changeless. In addition, sericin enhanced cell viability by the MTT assay. The above results showing the ability of sericin to promote protein production demonstrated its potential usefulness as a new biomaterial.

• Journal of Sericultural and Entomological Science
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• v.50 no.2
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• pp.76-81
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• 2012

Silk protein has been explored to be used for biomedical applications for several decades. However, it has not been used in this field cause to their irreversible crystallization after dissolving in water. The existing methods of silk protein hydrolysis using silkworm cocoon were used with harmful solvents and through a very complicated process. Therefore, we have developed novel methods for the production of water-soluble hydrolysate using silkworm gland. We manufactured two types of silkworm gland-derived hydrolysate (water-soluble SGH, SSGH; total SGH, TSGH) and compared the characteristics with commercial cocoon-derived sericin hydrolysate (CSH). The molecular weight of SGH ranged from 7 to 50 kDa (SSGH) and 5 to 15 kDa (TSGH) within glycine, alanine, and aspartic acid as a main amino acid composition. In contrast, CSH ranged from 15 to 50 kDa within serine and aspartic acid. The results of FTIR implied that SGH was more soluble form than CSH, as shown by the decrease in the ${\beta}$-sheet structure at $1630cm^{-1}$ on amide I peak. In comparison with 10% fetal bovine serum, 0.1% (1 mg/ml) SSGH had equivalent effect on the proliferation of human dermal fibroblasts and mesenchymal stem cells. All results of the SSGH made by novel manufacturing process indicate the SSGH is more preferable as a culture medium supplement than cocoon-derived sericin.