한국잠사곤충학회지 (Journal of Sericultural and Entomological Science)

한국잠사학회 (Korean Society of Sericultural Science)
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누에 실샘 가수분해물의 제조 및 특성 규명

Manufacture and Characterization of Silkworm Gland Hydrolysate

  • 황정욱 (차의과학대학교 바이오산업응용학과) ;
  • 이희삼 (농촌진흥청 국립농업과학원 잠사양봉소재과) ;
  • 김호진 (차의과학대학교 바이오산업응용학과) ;
  • 김규오 (차의과학대학교 바이오산업응용학과) ;
  • 최용수 (차의과학대학교 바이오산업응용학과)
  • Hwang, Jung Wook (Department of Applied Bioscience, CHA University) ;
  • Lee, Heui Sam (Sericultural & Apicultural Materials Division, National Academy of Agricultural Science, RDA) ;
  • Kim, Hojin (Department of Applied Bioscience, CHA University) ;
  • Kim, Kyu-Oh (Department of Applied Bioscience, CHA University) ;
  • Choi, Yong-Soo (Department of Applied Bioscience, CHA University)
  • 투고 : 2012.08.22
  • 심사 : 2012.10.09
  • 발행 : 2012.12.30
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초록

본 연구에서는 인체에 유해한 용매를 사용하지 않고 누에 실샘으로부터 가수분해물을 제조하는 새로운 공정을 개발하였다. 제조된 누에 실샘 가수분해물의 특성 분석을 통해 누에고치 유래 세리신 가수분해물(CSH)과 구성 성분 및 물성이 상이함을 확인하였다. 또한 1 mg/ml 수용성 누에 실샘 가수분해물(SSGH)을 첨가한 경우 10% FBS를 첨가한 대조군과 동등한 세포 증식효과가 있음을 증명하였다. 결론적으로 누에 실샘 가수분해물은 혈청을 대체할 수 있는 우수한 소재임을 확인하였다. 본 연구에서 개발한 제조방법을 통해 생산된 누에 실샘 가수분해물은 향후 화장품 및 의료용 소재로 그 응용범위를 확대할 수 있을 것으로 사료된다.

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.

키워드

참고문헌

  1. Cho SW, Kim JD, Lee KW, Choi DG, Han SK, Lee YJ, Cho HK (2006) Stimulation of insulin secretion and immune system in silk peptide and chitosan oligosaccharides. Kor J Gerontol 16, 88-93.
  2. Daithankar AV, Padamwar MN, Pisal SS, Paradkar AR, Mahadik KR (2005) Moisturizing efficiency of silk protein hydrolysate: Silk fibroin. Indian J Biotech 4, 115-121.
  3. Dash R, Mandal M, Ghosh SK, Kundu SC (2008) Silk sericin protein of tropical tasar silkworm inhibits UVB-induced apoptosis in human skin keratinocytes. Mol Cell Biochem 311, 111-119. https://doi.org/10.1007/s11010-008-9702-z
  4. Gamo T, Inokuchi T, Laufer H (1977) Polypeptides of fibroin and sericin secreted from the different section of the silk gland in Bombyx mori. Insect Biochem 7, 285-295. https://doi.org/10.1016/0020-1790(77)90026-9
  5. Gstraunthaler G (2003) Alternatives to the use of fetal bovine serum: serum-free cell culture. ALTEX 20, 275-281.
  6. Grzelak K (1995) Control of expression of silk protein genes. Biochem Mol Biol 110, 671-681. https://doi.org/10.1016/0305-0491(94)00215-G
  7. Hwang WJ, Jeong SN, Kim YS, Pi SH, You HK, Chung CP, Shin HS (2009) Clinical study of guided bone regeneration of extracted socket with PLA/PGA membrane and silk fibroin membrane. J Korean Acad Periodontol 39, 129-138.
  8. Jo YY, Kweon HY, Lee KG, Nam SH, Lee HS, Yeo JH (2011) The promotion of cell attachment and proliferation on silk fibroin. J Appl Biol Chem 54, 166-170. https://doi.org/10.3839/jabc.2011.028
  9. Kweon HY, Yeo JH, Kim KY, Kim YS, Song HS, Kim SJ, Woo SO, Han SM, Lee KG (2009) Characteristics of silk sericin extracted from sericinjam. Int J Indust Entomol 18, 121-124.
  10. Kim YA, Song EH, Shin KO, Lee YM, Cho YH (2012) Dietary effect of silk protein on epidermal levels of free sphingoid bases and phosphate metabolites in NC/Nga mice. Korean J Nutr 45, 113-120. https://doi.org/10.4163/kjn.2012.45.2.113
  11. Mondal M, Trivedy K, Kumar SN (2007) The silk protein, sericin and fibroin in silkworm, Bombyx mori Linn., - a review. Caspian J Env Sci 5, 63-76.
  12. Morikawa M, Kimura T, Murakami M, Katayama K, Terada S, Yamaguchi A (2009) Rat islet culture in serum-free medium containing silk protein sericin. J Hepatobiliary Pancreat Surg 16, 223-228. https://doi.org/10.1007/s00534-009-0049-y
  13. Minoura N, Aiba SI, Gotoh Y, Tsukada M, Imai Y (1995) Attachment and growth of cultured fibroblast cells on silk protein matrices. J Biomed Mater Res 29, 1215-1221. https://doi.org/10.1002/jbm.820291008
  14. Ohnishi K, Murakami M, Morikawa M, Yamaquchi A (2012) Effect of the silk protein sericin on cryopreserved rat islets. J Hepatobiliary Pancreat Sci 19, 354-360. https://doi.org/10.1007/s00534-011-0415-4
  15. Prasong S, Yaowalak S, Wilaiwan S (2009) Characteristics of silk fiber with and without sericin component: A comparison between Bombyx mori and philosamia ricini silks. Pakistan J Biol Sci 12, 872-876. https://doi.org/10.3923/pjbs.2009.872.876
  16. Sarovart S, Sudatis B, Meesilpa P, Brian P, Magaraphan R (2003) The use of sericin as an antioxidant and antimicrobial for polluted air treatment. Rev Adv Mater Sci 5, 193-198.
  17. Sasaki S, Nakagaki l (1980) Secretory mechanism of fibroin, a silk protein, in the posterior silk gland cells of Bombyx mori. Membr Biochem 3, 37-47. https://doi.org/10.3109/09687688009063877
  18. Takahashi M, Tsujimoto K, Yamada H, Takagi H, Nakamori S (2003) The silk protein, sericin, protects against cell death caused by acute serum deprivation in insect cell culture. Biotechnol Lett 25, 1805-1809. https://doi.org/10.1023/A:1026284620236
  19. Takasu Y, Yamada H, Tsubouchi K (2002) Isolation of three main sericin components from the cocoon of the silkworm, Bombyx mori. Biosci Biotechnol Biochem 66, 2715-2718. https://doi.org/10.1271/bbb.66.2715
  20. Terada S, Nishimura T, Sasaki M, Yamada H, Miki M (2002) Sericin, a protein derived from silkworms, accelerates the proliferation of several mammalian cell lines including a hydridoma. Cytotechnology 40, 3-12. https://doi.org/10.1023/A:1023993400608
  21. Tokutake S (1980) Isolation of the smallest component of silk protein. Biochem J 187, 413-417. https://doi.org/10.1042/bj1870413

피인용 문헌

  1. Whitening Effect of Storage Protein 2 from Silkworm Hemolymph vol.05, pp.09, 2014, https://doi.org/10.4236/abb.2014.59089