• Title/Summary/Keyword: silk fibroin 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.

Silk Fibroin Microsphere and Its Characterization

  • Yeo, Joo-Hong;Lee, Kwang-Gill;Lee, Yong-Woo;Kweon, Hae-Yong;Woo, Soon-Ok
    • 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.

Anti-adipogenic Effect of Hydrolysate Silk Fibroin in 3T3-L1 Cells

  • Chon, Jeong-Woo;Lee, Kwang-Gill;Park, Yoo-Kyoung;Park, Kyung-Ho;Yeo, Joo-Hong
    • International Journal of Industrial Entomology and Biomaterials
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    • v.21 no.2
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    • pp.169-174
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    • 2010
  • Hydrolysate silk fibroin (HSF) is a fibrous protein composed of parallel $\beta$-structures and is made from pure silk elements including 18 amino acids, with glycine, alanine, and serine comprising of over 80% of the amino acids. Numerous studies have documented a range of effects of HSF, including moisturizing, antioxidant activity, nervous system disorders, and many more. We investigated whether HSF has anti-obesity effects in vitro. The effects of HSF inhibition on lipid accumulation and acceleration of lipid degradation in 3T3-L1 cells were studied. Treatment of 3T3-L1 cells with HSF caused significant inhibition of cell viability, an increase in glycerol release, and a decreased in adipocyte differentiation. Moreover HSF stimulated downregulated of adipogenic enzyme expressions (PPAR${\gamma}$ and C/EBP${\alpha}$) and up-regulated of fatty oxidation enzyme expressions (CPT-1 and UCP-2). Based on these results, hydrolysate silk fibroin can be suggested as a potential therapeutic substance as part of a prevention or treatment strategy for obesity.

Synthesis and Characterization of Acrylic Polymer Containing Silk Protein

  • Zhongmin Chen;Kim, Mutsumi ura;Masahiro Suzuki;Yoshiyuki Kondo;Kenji Hanabusa;Hirofusa Shirai
    • Proceedings of the Korean Fiber Society Conference
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    • 2003.10a
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    • pp.87-87
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    • 2003
  • Three kinds of acrylic polymers containing silk protein were synthesized, which are (1) blending of silk fibroin (SF) fiber and polyacrylonitrile (PAN); (2) graft-copolymer of PAN onto SFs; (3) random-copolymer synthesized by copolymerization of acrylonitrile (AN) and silk fibroin peptide (SFP) with vinyl groups, and their solubility, thermal property, and moisture absorption was investigated, respectively. These polymers have difference solubility and attributable to their structure. Their excellent thermal stabilities and better moisture absorptions were indicated.

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Effects of Silk Fibroin Powder on Oxygen Radicals and Their Scavenger Enzymes in Serum of Rats (실크 피브로인 분말이 혈청중의 활성산소 및 제거효소에 미치는 영향)

  • 최진호;김대익;박수현;김동우;이종수;이용우
    • Journal of Sericultural and Entomological Science
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    • v.41 no.3
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    • pp.216-221
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    • 1999
  • This study was designed to investigate the effects of silk fibroin powder on oxygen radicals and their scavenger enzymes in serum of rats. Sprague-Dawley(SD) male rats (160${\pm}$10 g) were fed experimental diets (SFP-2.5 and SFP-5.0 groups)added 2.5 and 5.0 g/kg BW/day for 6 weeks. Triglyceride levels were remarkably inhibited (15∼25%) in SFP-2.5 and SFP-5.0 groups compared with control group. Hydroxyl radical ($.$OH) formations resulted in a marked decrease (15%∼25%) compared with control group, while superoxide radical and hydrogen peroxide formations resulted in a significant decrease (6∼12% and 10%). Lipid peroxide and oxidized protein (>C=O group) productions resulted in a significant decrease (6∼12% and 6%) compared with control group. Superoxide dismutase (SOD) and catalase (CAT) activities were remarkably increased (35∼55% and 40∼50%), but glutathione peroxidase (GSHPX) activities were significantly increased (13%) compared with control group. These results suggest that anti-aging effect of silk fibroin powder (SFP) may play a pivotal role in attenuating a various age-related changes.

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Effect of degumming conditions on the fluorescence intensity of fluorescent silk cocoons: A combined experimental and molecular dynamics study

  • Chan Yeong, Yu;Ezekiel Edward, Nettey-Oppong;Elijah, Effah;Su Min, Han;Seong-Wan, Kim;Seung Ho, Choi
    • International Journal of Industrial Entomology and Biomaterials
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    • v.45 no.2
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    • pp.56-69
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    • 2022
  • Silk is a unique natural biopolymer with outstanding biocompatibility, high mechanical strength, and superior optical transparency. Due to its excellent properties, silk has been widely reported as an ideal biomaterial for several biomedical applications. Recently, fluorescent silk protein, a variant of native silk, has been reported as a biophotonic material with the potential for bioimaging and biosensing. Despite the realization of fluorescent silk, the traditional degumming process of fluorescence silk is crude and often results in fluorescence loss. The loss of fluorescent properties is attributed to the sensitivity of silk fibroin to temperature and solvent concentration during degumming. However, there is no comprehensive information on the influence of these processing parameters on fluorescence evolution and decay during fluorescent silk processing. Therefore, we conducted a spectroscopic study on fluorescence decay as a function of temperature, concentration, and duration for fluorescent silk cocoon degumming. Sodium carbonate solution was tested for degumming the fluorescent silk cocoons with different concentrations and temperatures; also, sodium carbonate solution is combined with Alcalase enzyme and triton x-100 to find optimal degumming conditions. Additionally, we conducted a molecular dynamics study to investigate the fundamental effect of temperature on the stability of the fluorescent protein. We observed degumming temperature as the prime source of fluorescent intensity reduction. From the MD study, fluorescence degradation originated from the thermal agitation of fluorescent protein Cα atoms and fluctuations of amino acid residues located in the chromophore region. Overall, degumming fluorescent silk with sodium carbonate and Alcalase enzyme solution at 25 ℃ preserved fluorescence.

Production of the yellow fluorescent silk using the fibroin heavy chain protein expression system in transgenic silkworm (피브로인 H-chain 재조합 단백질 발현시스템을 이용한 황색형광실크의 제작)

  • Kim, Seong Wan;Choi, Kwang-Ho;Kim, Seong Ryul;Yun, Eun Young;Park, Seung Won;Kang, Seok Woo;Goo, Tae Won
    • Journal of Sericultural and Entomological Science
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    • v.52 no.2
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    • pp.102-109
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    • 2014
  • We constructed the fibroin H-chain expression system to produce enhanced yellow fluorescent proteins (EYFP) in the silk of transgenic silkworm. Fluorescent silk could be made by fusing EYFP cDNA to the heavy chain gene and injecting it into a silkworm. The EYFP 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 EYFP/H-chain fusion gene was regulated by the fibroin H-chain promoter. The yellow fluorescence proving that the fusion protein was present in the silk. Accordingly, we suggest that the EYFP fluorescence silk will enable the production of novel biomaterial based on the transgenic silk.

Physical Properties of Silk Fibroin Films Treated with Various Plasticizers

  • Ma, Yu-Hyun;Song, Kyung-Bin
    • Preventive Nutrition and Food Science
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    • v.10 no.2
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    • pp.187-190
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    • 2005
  • Silk fibroin (SF) films were prepared using various plasticizers and their physical properties were determined. Polyethylene glycol (PEG)-SF and polypropylene glycol (PPG)-SF films had tensile strengths (TS) of 23.71 MPa and 24.51 MPa, respectively, whereas the glycerol (G)-SF film had the lowest TS of 14.24 MPa. G-SF film had the highest $\%$ elongation, compared to PPG-SF and PEG-SF films. Water vapor permeability (WVP) of SF films varied with addition of plasticizers, and PEG-SF film had the lowest WVP. There was no significant difference in Hunter L value among treatments, but PEG-SF film had higher Hunter a and b values. These results suggest that SF film could be applied to food packaging and that the addition of plasticizers should improve the physical properties of SF film.

Effects of PEG (Polyethylene Glycol) Concentration and Mixing Ratio of PEG/Gly (Glycerol) on the Physical Properties of Silk Fibroin Films (PEG(polyethylene glycol) 농도와 PEG/Gly(glycerol) 흔합비에 따른 견 피브로인 필름의 물성)

  • Ma, Yu-Hyun;Song, Kyung-Bin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.1
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    • pp.121-125
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    • 2006
  • To study the effects of plasticizer concentration and its ratio on the physical properties of silk fibroin films, polyethylene glycol (PEG) was used at 4 different concentrations; 1, 2, 3, $4.5\%$ (w/v). Tensile strength (TS) and water vapor permeability (WVP) increased with the increase of PEG concentration, while opposite trend was observed for percent elongation of silk fibroin films. WVP of silk fibroin films increased from $2.54\;ng{\cdot}m/m^2spa$ for $1\%$ of PEG to $5.41\;ng{\cdot}m/m^2sPa$ for $4.5\%$. In addition, a mixture of PEG and glycerol (Gly) as a plasticizer was used at the ratio of 100:0, 75:25, 50:50, 25:75, and 0:100 (w/w). Percent elongation of the films was improved to $130.95\%$ when the ratio of 75:25 was used. On the contrary, WVP of silk fibroin films increased with the decrease of the ratio of PEG:Gly. Effect of the plasticizer concentration and its ratio on the color of silk fibroin films was negligible. These results suggest that mixture of PEG and Gly as a plasticizer provide more flexible than PEG alone in silk fibroin films, and the best ratio of PEG to Gly was 75:25.

The Degumming and Sericin Recovery of the Silk fabric Using the Electrolytic Water (전해수를 이용한 견섬유 정련 및 세리신 회수 (I))

  • 배기서;하헌주;박광수
    • 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.