• Title/Summary/Keyword: Silk protein

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Silk Protein as a Fetal Bovine Serum Substitute for Animal Cell Culture

  • Jo, You-Young;Kweon, HaeYong;Ji, Sang Deok;Kim, Jong Gil;Kim, Kee Young
    • Microbiology and Biotechnology Letters
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    • v.47 no.4
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    • pp.487-497
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    • 2019
  • Fetal Bovine Serum (FBS) is an essential substance added to animal cell culture medium. However, its composition is unclear causing problems such as development of an immune response when cultured cells are transplanted into the human body. In this study, silk sericin, silk fibroin, and hemolymph obtained from silkworms were added to the cell culture medium in order to determine if it can replace FBS. After establishment of the cell culture, cell proliferation and expression levels of cell growth-related genes were compared with those of control cells (cells cultured in the medium with 10% FBS). Results showed that the test group treated with silk fibroin extracted from a Korean silkworm variety, Kumokjam could replace 10% FBS. In addition, expression levels of cell growth related genes such as Fibronectin and TGF-β1 increased significantly in cells cultured using silk fibroin, depending on the concentration used in cell adhesion and cell proliferation [24]. To date, no studies have been conducted to find a replacement for FBS. Thus, this study was carried out to develop a substitute for FBS by using silkworm-derived alternatives such as silkworm hemolymph, silk sericin, and silk fibroin, which are cheap and have various physiological effects, cell promoting effects, and can be mass produced.

Production of fluorescent green silk using fibroin H-chain expression system (피브로인 H-chain 재조합 단백질 발현시스템을 이용한 녹색형광실크 생산)

  • Kim, Seong Wan;Yun, Eun Young;Choi, Kwang-Ho;Kim, Seong Ryul;Park, Seung Won;Kang, Seok Woo;Goo, Tae Won
    • Journal of Sericultural and Entomological Science
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    • v.51 no.2
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    • pp.153-158
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    • 2013
  • To express green fluorescent protein in the cocoon of silkworm, we constructed the fibroin H-chain expression system to produce enhanced green fluorescent protein (EGFP) in the cocoon of transgenic silkworms. The EGFP 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 EGFP/H-chain fusion gene was regulated by the fibroin H-chain promoter. The use of the 3xP3-driven DsRed2 cDNA as a marker allowed us to rapidly distinguish transgenic silkworm. A mixture of the donor and helper vector was micro-injected into 1,200 eggs of bivoltin silkworms, Baegokjam. We obtained 8 broods. The cocoon displayed strong green fluorescence, proving that the fusion protein was present in the cocoon. Also, the presence of fusion proteins in cocoons was demonstrated by SDS-PAGE and immunoblotting. Accordingly, we suggest that the EGFP fluorescence silk will enable the production of the novel biomaterial based on the transgenic silk.

Dietary Effect of Silk Protein on Colon Cancer of Animal (실크 단백질 식이공급이 동물대장암에 미치는 영향)

  • Yeo, Joo-Hong;Lee, Kwang-Gill;Woo, Soon-Ok;Kweon, Hae-Yong;Han, Sang-Mi;Park, Kyung-Ho;Kim, Sung-Su
    • Journal of Sericultural and Entomological Science
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    • v.49 no.2
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    • pp.67-70
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    • 2007
  • This study was done for the dietary effect of silk protein, molecular controlled silk sericin and silk fibroin, on colon cancer of animal. The ICR mice were given weekly injections of 1,2-dimethylhydrazine (DMZ) for the initial 10 mg/kg with four weeks, and fed a diets supplemented with 3% (v/v) molecular controlled sericin and fibroin for 6 weeks, respectively. Then, the body weight, efficiency of dietary, typical histologic appearance of animal colon tissue and COX-2 effect were studied, too. Molecular controlled sericin protein are shown more higher dietary effect on animal colon cancer than fibroin supplemental case. The results suggest a potential usefulness of sericin as a chemopreventive for colon carcinogenesis and the development functional food.

High maysin corn silk extract reduces body weight and fat deposition in C57BL/6J mice fed high-fat diets

  • Lee, Eun Young;Kim, Sun Lim;Kang, Hyeon Jung;Kim, Myung Hwan;Ha, Ae Wha;Kim, Woo Kyoung
    • Nutrition Research and Practice
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    • v.10 no.6
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    • pp.575-582
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    • 2016
  • BACKGROUNG/OBJECTIVES: The study was performed to investigate the effects and mechanisms of action of high maysin corn silk extract on body weight and fat deposition in experimental animals. MATERIALS/METHODS: A total of 30 male C57BL/6J mice, 4-weeks-old, were purchased and divided into three groups by weight using a randomized block design. The normal-fat (NF) group received 7% fat (diet weight basis), the high-fat (HF) group received 25% fat and 0.5% cholesterol, and the high-fat corn silk (HFCS) group received high-fat diet and high maysin corn silk extract at 100 mg/kg body weight through daily oral administration. Body weight and body fat were measured, and mRNA expression levels of proteins involved in adipocyte differentiation, fat accumulation, fat synthesis, lipolysis, and fat oxidation in adipose tissue and the liver were measured. RESULTS: After experimental diet intake for 8 weeks, body weight was significantly lower in the HFCS group compared to the HF group (P < 0.05), and kidney fat and epididymal fat pad weights were significantly lower in the HFCS group compared to the HF group (P < 0.05). In the HFCS group, CCAAT/enhancer binding protein-${\beta}$, peroxisome proliferator-activated receptor-${\gamma}1$ (PPAR-${\gamma}1$), and PPAR-${\gamma}2$ mRNA expression levels were significantly reduced (P < 0.05) in the epididymal fat pad, whereas cluster of differentiation 36, lipoprotein lipase, acetyl-CoA carboxylase-1, sterol regulatory element binding protein-1c, pyruvate dehydrogenase kinase, isozyme-4, glucose-6-phosphate dehydrogenase, and stearoyl-CoA desaturase-1 mRNA expression levels were significantly decreased in liver and adipose tissues (P < 0.05). In the HFCS group, mRNA expression levels of AMP-activated protein kinase, hormone-sensitive lipase, and carnitine palmitoyltransferase-1 were elevated (P < 0.05). CONCLUSIONS: It can be concluded that high maysin corn silk extract inhibits expression of genes involved in adipocyte differentiation, fat accumulation, and fat synthesis as well as promotes expression of genes involved in lipolysis and fat oxidation, further inhibiting body fat accumulation and body weight elevation in experimental animals.

Single-dose oral toxicity study of genetically modified silkworm expressing EGFP protein in ICR mouse

  • Jang, Kyung-Min;Kim, Sung-Gun;Park, Ji-Young;Choi, Won-Ho;Lee, Jae-Woo;Jegal, Hyeon-Young;Kweon, Soon-Jong;Choi, Kwang-Ho;Park, Jung-Ho
    • 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.

Application for Dietary Resources by Silk Protein (실크 단백질의 식이 소재로서의 응용)

  • Yeo, Joo-Hong;Lee, Kwang-Gill;Kweon, Hae-Yong;Han, Sang-Mi;Park, Kyung-Ho;Kim, Sung-Su;Shin, Bong-Sub
    • Journal of Sericultural and Entomological Science
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    • v.48 no.1
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    • pp.6-10
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    • 2006
  • Recently, B. mori proteins such as silk fibroin and silk sericin have been found to have a water-holding capacity, anti hydrogen peroxide toxicity, antioxidant activity and tyrosinase-inhibitory activity (Yeo 2006, Kurioka 1999 & 2004), implying its potential usefulness of the application for cosmetic and functional food(Yamazaki 1999 & Une 2000). We are tried to application for dietary resources of B. mori silk fibroin and sericin that were prepared to some of different molecular cutting these resources by preparative recycling HPLC system. In our studies with rats have demonstrated that consumption of these silk proteins are being prevents constipation effect and it is maybe enhances intestinal absorption of water and dietary effects. These some of useful results further suggest a usefulness of sericin as dietary resources for health.