• Journal of Life Science
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• v.10 no.5
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• pp.511-518
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• 2000

This study was designed to investigate the effects of silk fibroin powder (SFP : Mw 500) on oxidative stress and membrane fluidity in brain membranes of rats. Sprague-Dawley (SD) male rats (160$\pm$10 g) were fed basic diet (control group), and experimental diets (SFP-2.5 and SFP-5.0 groups) added 2.5 and 5.0 g/kg BW/day for 6 weeks. Cholesterol level was significantly decreased about 8.0% in brain microsomes of SFP-5.0 group only compared with control group. Membrane fluidities were significantly increased (12.9% and 15.2%, respectively) in brain microsomes of SFP-2.5 and SFP-5.0 groups, but significant difference between in brain mitochondria of these two groups could be not obtained. Basal oxygen radicals (BOR) in brain mitochondria and microsomes were significantly ingibited (10.4%, and 24.0%, 7.9% and 14.9%, respectively) by SFP-2.5 and SFP-5.0 groups compared with control group. Induced oxygen radicals (IOR) in brain mitochondria and microsomes were significantly inhibited (11.8% and 14.1%, respectively) by SFP-5.0 groups compared with control group compared with control group. Lipid peroxide (LPO) levels were dose-dependently decreased (12.9% and 21.9%, 13.2% and 22.5%, respectively) in brain mitochondria and microsomes of SFP-2.5 and SFP-5.0 groups compared with control group. Oxidized protein (OP) levels were significantly decreased (15.7% and 17.1%, 16.7% and 15.7%, respectively) in brain mitochondria and microsomes of SFP-2.5 and SFP-2.5 and SFP-5.0 groups compared with control group. These results suggest that administration of SFP may play an effective role in a attenuating a oxidative stress and increasing a membrane fluidity in brain membranes.

• KSBB Journal
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• v.13 no.1
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• pp.114-118
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• 1998

The effects of fibron solubilization conditions on molecular weight distribution of enzymatically-hydrolyzed silk peptides were investigated. The weight-averaged molecular weights of silk proteins prepared by solubilization with calcium chloride, ethylenediamine and sulfuric acid were 41600, 3308, and 1268 dalton, respectively. Silk peptides in the average molecular weight range of 600-1200 dalton were obtained by protease treatment from solubilized silk fibroin. After the acid hydrolysis of silk protein using hydrochloric acid for 24 hr, silk protein was hydrolyzed to peptides whose average molecular weight and free amino acid content were 145 dalton and 80%, respectively. It was possible to control molecular weight distribution of silk peptides by the combination of solubilization and hydrolysis methods. Among the various treatment methods, acid solubilization followed by protease treatment had an advantage of molecular weight control for the peptide production.

• Proceedings of the Korean Society of Sericultural Science Conference
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• 2003.10a
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• pp.55-56
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• 2003

Silk fibroin (SF) is one of the typical protein polymer produced by silkworm, Bombyx mori. SF has been used as textile fiber and surgical suture fur thousands of years due to its unique gloss, handle, and mechanical properties. Recently, SF has been intensively studied to diverse usage for biotechlological and biomedical fields because of their reproducibility, environmental compatibility, non-toxicity, and biological compatibility. Based on its biocompatibility, the possible uses of regenerated SF have been proposed including substrate for cell culture[1], enzyme immobilization[2], and matrix for drug release[3]. (omitted)

• Journal of Microbiology and Biotechnology
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• v.22 no.4
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• pp.494-500
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• 2012

Silk fibroin (SF) peptide has been traditionally used as a treatment for flatulence, spasms, and phlegm. In this study, we examined whether SF peptide enhanced the anti-inflammatory effect of PEP-1-FK506 binding protein (PEP-1-FK506BP) through comparing the anti-inflammatory activities of SF peptide and/or PEP-1-FK506BP. In the presence or absence of SF peptide, transduction levels of PEP-1-FK506BP into HaCaT cells and mice skin and anti-inflammatory activities of PEP-1-FK506BP were identified by Western blot and histological analyses. SF peptide alone effectively reduced both mice ear edema and the elevated levels of cyclooxygenase-2, interleukin-6 and $-1{\beta}$, and tumor necrosis factor-${\alpha}$, showing similar anti-inflammatory effect to that of PEP-1-FK506BP. Furthermore, co-treatment with SF peptide and PEP-1-FK506BP exhibited more enhanced anti-inflammatory effects than the samples treated with SF peptides or PEP-1-FK506BP alone, suggesting the possibility that SF peptide and PEP-1-FK506BP might interact with each other. Moreover, the transduction data demonstrated that SF peptide did not affect the transduction of PEP-1-FK506BP into HaCaT cells and mice skin, indicating that the improvement of anti-inflammatory effect of PEP-1-FK506BP was not caused by enhanced transduction of PEP-1-FK506BP. Thus, these results suggest the possibility that co-treatment with SF peptide and PEP-1-FK506BP may be exploited as a useful therapy for various inflammation-related diseases.

• Journal of Nutrition and Health
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• v.39 no.6
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• pp.520-528
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• 2006

Free amino acids in epidermis function as a major component of Natural Moisturizing Factor (NMF), which maintains the optimal level of water in skin even at the low humidity. In fact, the depletion of free amino acids is reported in the epidermis of atopic dermatitis, the skin condition involving dryness. As an effort searching the dietary source for improving the level of water and free amino acid in epidermis, the dietary effects of silk protein, sericin (S) and fibroin (F) on trans epidermal water loss (TEWL), and plasma and epidermal levels of free amino acids were compared in this study. Thirty of male NC/Nga mice, an animal model of atopic dermatitis, were divided into three groups: group CA as an atopic control with control diet, group S: 1% sericin diet and group F: 1% fibroin diet. Ten of male BALB/c mice were served as group C (control group) with control diet. All mice were fed on diet and water ad libitum for 10weeks. Dry skin condition was established in group CA as TEWL was increased (148.7% of group C). In parallel, epidermal level of glutamate, one of major amino acids functioning as NMF, was dramatically decreased and epidermal levels of methionine and alanine were inversely elevated. Dietary supplementation of sericin (group S) reduced TEWL at the similar level with group C and increased epidermal levels of glutamate as well as serine and glycine, the other major amino acids as NMF. Despite a marked decrease of methionine and alanine, the reduction of TEWL and epidermal levels of glutamate, serine and glycine of group F were less than of group S. Furthermore, in contrast to similar levels of other free amino acids in plasma and epidermis of group S and group C, plasma and epidermal levels of other free amino acids, specifically phenylalanine, isoleucine, cysteine and tyrosine in epidermis of group F, were significantly higher than of group C. Together, our data demonstrate that dietary supplementation of sericin is more effective at improving dry skin condition that paralleled with the normalization of free amino acids in plasma and epidermis of NC/Nga mice.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.34 no.1
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• pp.1-11
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• 2012

Purpose: The purpose of this study was to evaluate the bone regeneration capacity of silk fibroin (SF) when combined with beta tricalcium phosphate (${\beta}$-tricalcium phosphate [TCP]) and rh-bone morphogenetic protein (BMP) in vivo by micro-computed tomography (CT), soft x-ray, and histological analysis. Methods: A total of 56 critical size defects formed by a trephine bur made on 28 adult female Spague-Dawley rats were used for this study and the defect size was 5.0 mm in diameter. The defects were transplanted with (1) no graft material (raw defect), (2) autogenous bone, (3) SF ($10{\mu}g$), (4) SF-BMP ($10{\mu}g$, $0.8{\mu}g$ each), and (5) SF+${\beta}$-TCP ($10{\mu}g$). At 4 and 8 weeks after operation, the experimental animals were sacrificed. Samples were evaluated with soft x-ray, histological examinations and 3-dimensional micro-CT analysis. Results: In the 3-dimensional micro-CT evaluation, bone volume and bone surface data were higher in the SF-BMP ($12.8{\pm}1.5$, $138.6{\pm}45.0$ each) (P<0.05) and SF-TCP ($12.3{\pm}1.5$, $144.9{\pm}30.9$ each) group than in the SF group ($6.1{\pm}3.3$, $77.2{\pm}37.3$ each) (P<0.05), except for the autogenous group ($15.0{\pm}3.0$, $190.7{\pm}41.4$ each) at 4 weeks. At 8 weeks, SF-BMP ($16.8{\pm}3.5$, $173.9{\pm}34.2$ each) still revealed higher (P<0.05) bone volum and surface, but SF-TCP ($11.3{\pm}1.5$, $1132.9{\pm}52.1$ each) (P=0.5, P=0.2) revealed the same or lower amount compared with the SF group ($13.8{\pm}2.7$, $127.5{\pm}44.8$ each). The % of bone area determined by radiodensity was higher in the SF-TCP ($31.4{\pm}9.1%$) and SF-BMP ($36.2{\pm}16.2%$) groups than in the SF ($19.0{\pm}10.4$) group at the period of 4 weeks. Also, in the histological evaluation, the SF-BMP group revealed lower inflammation reaction, lower foreign body reaction and higher bone healing than the SF group at postoperative 4 weeks and 8 weeks. The SF-TCP group revealed lower inflammation at 4 weeks, but accordingly, as the TCP membrane was absorbed, inflammatory and foreign body reaction are increased at 8 weeks. Conclusion: The current study provides evidence that the silk fibrin can be used as an effective grafted material for tissue engineering bone generation through a combination of growth factor or surface treatment.

• Development and Reproduction
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• v.24 no.3
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• pp.231-239
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• 2020

Many benefits of silk protein fibroin (SPF) have been suggested in biomedical applications; and notably, significant SPF effects have been observed for metabolic syndromes that are directly linked to insulin resistance, such as type 2 diabetes mellitus (T2DM). Based on our previous findings, we believe that SPF from spiders exhibits outstanding glucose-lowering effects in diabetic BKS.Cg-m+/+Lepr^db mice. In order to evaluate the dietary effects of SPF in diabetic animals, we generated several lines of transgenic rice (TR) that expresses SPF, and the feeding of TR-SPF to diabetic animals decreased blood glucose levels, but did not change insulin levels. Western blot analyses of hepatic proteins showed that AMP-activated protein kinase (AMPK) expression and phosphorylation both decreased in TR-SPF-fed groups, compared with controls. This finding suggests that the glucose-lowering effects in this diabetic animal model might be AMPK-independent. In contrast, six-transmembrane protein of prostate 2 (STAMP2) was upregulated after TR-SPF exposure. Together with STAMP2, the Akt protein phosphorylation increased after TR-SPF exposure, which indicates that STAMP2 leads to Akt phosphorylation and thus increases insulin sensitivity in hepatocytes. Importantly, the hepatic steatosis that was seen in the liver of diabetic mice was remarkably alleviated in TR-SPF-fed mice. Hepatocytes that were immunopositive for STAMP2 were overwhelmingly observed in hepatic tissues from TR-SPF-fed mice compared to the control. Taken together, these results suggest that feeding diabetic mice with TR-SPF upregulates STAMP2 expression and increases Akt phosphorylation in hepatic tissues and thus potentially alleviates insulin resistance and hepatic steatosis.

• Journal of Sericultural and Entomological Science
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• v.42 no.1
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• pp.58-64
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• 2000

This study was designed to investigate the effects of silk fibroin powder (Mw 500) on oxidative stress and membrane fluidity in liver membranes of rats. Sprague-Dawley (SD) male rats (160$\pm$10g) were fed basic diet (control group), and experimental diets (SEP-2.5 and SFP-5.0 groups) added 2.5 and 5.0 g/kg BW/day for 6 weeks. Cholesterol levels resulted in a significant decrease (12.1% and 9.0%, respectively) in the liver mitochondria and microsomes of SEP-5.0 group compared with control group. Membrane fluidity as significantly increased (16.1% and 16.5%, 5.8% and 17.4%) in the liver mitochondria and microsomes were significantly inhibited (16.1% and 18.3%, 8.1% and 15.1%, respectively) at the SFP-2.5 and SEP-5.0 groups compared with control group. Induced oxygen radicals (BOR) in liver mitochondria and microsomes were significantly inhibited (16.1% and 18.3%, 8.1% and 15.1%, respectively) at the SFP-2.5 and SEP-5.0 groups compared with control group. Induced oxygen radicals (IOR) in liver microsomes were significantly inhibited (17.0% and 26.6%, respectively) at the SFP-2.5 and SFP-5.0 groups compared with control group, but IOR in liver mitochondria was significantly inhibited about 12.3% at the SWP-400 group only compared with control group. Lipid peroxide (LPO) levels were significantly decreased (8.3% and 18.0%, 13.4% and 18.4%, respectively) in the liver mitochondria and microsomes of SFP-2.5 and SFP-5.0 groups compared with control group. Oxidized protein (OP) levels were dose-dependently decreased (5.4% and 11.6%, 19.0% and 24.4%, respaectively) in the iver mitochondria and microsomes of SFP-2.5 and SFP-5.0 groups compared with control group. These results suggest that administration of SFP may play an effective role in attenuating an oxidative stress and increasing a membrane fluidity in liver membranes.

• KSBB Journal
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• v.24 no.5
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• pp.458-462
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• 2009

Silk fibroin is a structural protein from Bombyx mori and can be sulfated to provide biofunctional polypeptides showing antiviral and anticoagulating activities. However, the sulfated fibroins have not been characterized enough to present their compositions and structures. In this report, sulfation reaction was investigated by varying the reaction temperature and sulfuric acid concentration and analysis of the resulting peptides were carried out. The degree of sulfation was proportion to sulfuric acid concentration but the maximum product yield was obtained at $60^{\circ}C$ and 5% of sulfuric acid concentration. FT-IR spectrum, UV absorption and NMR spectrum support that o-sulfate ester was formed at the hydroxyl group of serine and tyrosine residues. Size exclusion chromatography identified that sulfated fibroin was composed of a highly hydrolyzed polypeptide mixtures and peptides of relatively higher molecular weight.

• Korean Journal of Agricultural Science
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• v.43 no.1
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• pp.109-115
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• 2016

Silk has had a reputation as a luxurious and sensuous fabric but it is not popular due to the expensive price and poor durability. To develop the silk materials that apply the various industries, the artificially synthesized gene can be introduced into the silkworm and expressed in the silk gland. Transgenic silkworms for the mass production of green fluorescent silks are generated using a fibroin H-chain expression system. For commercial use, safety assessment of the transgenic silkworms is essential. The purpose of this study was to examine the potential acute oral toxicity of EGFP protein expressed in genetically modified (GM) fluorescence silkworm and to obtain the approximative lethal dose in the male and female at 6-weeks ICR mice. EGFP protein was fed at a dose of 2,000 mg/kg body weight in five male or five female mice. Mortalities, clinical findings and body weight changes were monitored for 1, 3, 7, 14 days after dosing. At the end of 14 day observation period, all mice were sacrificed, and the postmortem necropsy were performed. The test group was not observed death case. Also the effect was not admitted by test substance administration in common symptoms, the body weight and postmortem. The results of single-dose oral toxicity test showed that approximative lethal dose of EGFP protein expressed in fluorescence silkworm was considered to exceed the 2,000 mg/kg body weight in both sexes.