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Stimulation of Nitric Oxide Production in RAW 264.7 Macrophages by the Peptides Derived from Silk Fibroin.  

박금주 (한동대학교 대학원 생명공학과)
현창기 (한동대학교 생물식품공학부)
Publication Information
Microbiology and Biotechnology Letters / v.30, no.1, 2002 , pp. 39-45 More about this Journal
Abstract
It was found that the peptides originated from the hydrolysates of silk fibroin have in vitro immunostimulating effects in murine macrophage RAW264.7 cells. The stimulation effects on nitric oxide (NO) production resulted from treatments of acid or enzymatic hydrolysates were measured. The silk fibroin preparation isolated from cocoon was most efficiently digested by acid hydrolysis. Even though the sole treatment of acid hydrolysate stimulated the NO production in dose-dependent pattern, a part of its activity was found to be caused by the contaminated endotoxin, LPS. When each endotoxin-free hydrolysates obtained by filtering it through an ultrafiltration membrane of molecular weight (MW) cut-off 10,000 to eliminate LPS was used, the peptic hydrolysate with lowest degree of hydrolysis showed the highest activity. The fractions of peptic hydrolysate with MW ranges of 1,000∼10,000, 500∼1,000 and below 500 also showed a higher MW-higher activity correlation. From the analyses of amino acid composition of each hydrolysate, it was found that the contents of arginine, lysine, alanine and glycine residues affected the activity level of hydrolysate. The results of this study showed a possibility of utilizing fibroin as a source for immunostimulating (chemopreventive) functional peptides.
Keywords
Silk fibroin; Peptides; RAW 264.7 macrophages; Nitric oxide production;
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1 Akai, H. 1999. New physiological functions of silk material. Shokuhin to Kaihatsu 34: 45-47.
2 Kim, M. K.., K. H. Lee, H. J. Lim, S. J. Lee, S. H. Lee, and K. S. Min. 1996. Preparation protocols for the functional polypeptide materials from cocoon. Korean Patent 98712.
3 Urban, J. L., H. M. Shepard.T, L. Rothstein, B. J. Sugarman, and H. Schreiber. 1986. Tumor necrosis factor: a potent effector molecule for tumor cell killing by activated macrophages. Proc. Natl. Acad. Sci. USA 83: 5233-5237.
4 Kayser, H. and H. Meisel. 1996. Stimulation of human peripheral blood lymphocytes by bioactive peptides derived from bovine milk proteins. FEBS Lett. 383: 18-20.   DOI   PUBMED   ScienceOn
5 Sohn, D. H. 1997. Bioactive peptides enhancing health and their applications. Food Sci. Industry 30: 22-29.
6 Yoshikawa, M., K. Kishi, M. Takahashi, A Watanabe, T. Miyamura, M. Yamazaki, and H. Chiba. 1993. Immunostimulating peptide derived from soybean protein. Ann. N. Y. Acad. Sci. 685: 375-376.
7 Chen, K., Y. Umeda, and K. Hirabayashi. 1996. Enzymatic hydrolysis of silk fibroin. J. Seric. Sci. Jpn. 65: 131-133.
8 Ding, A. H., C. F. Nathan, and D. J. Stuehr. 1988. Release of reactive nitrogen intermediates and reactive oxygen intermediates from mouse peritoneal macrophages. J. Immunol. 141: 2407-2412.
9 Carmichael, J., W. G. DeGraff, A F. Gazdar, J. D. Minna, and J. B. Mitchell. 1987. Evaluation of a tetrazolium-based semiautomated colorimetric assay: assessment of chemosensitivity testing. Cancer Res. 47: 936-942.
10 Qian J., Y. Liu, H. Liu, T. Yu, and J. Deng. 1997. Immobilization of horseradish peroxidase with a regenerated silk fibroin membrane and its application to a tetrathiafulvalenemediating H202 sensor. Biosens. Bioelectron. 12: 1213-1218.
11 Mantovani, A., B. Bottazzi, F. Colotta, S. Sozzani, and L. Ruco. 1992. The origin and function of tumor-associated macrophages. Immuno!. Today 13: 265-270.
12 Remillard, J. F., M. C. Gould, P. F. Roslansky, and T. J. Novitsky. 1987. Quantitation of endotoxin in products using the LAL kinetic turbidimetric assay. Prog. Clin. Biol. Res. 231: 197-210.
13 Minoura N., S. I. Aiba, M. Higuchi, Y. Gotoh, M. Tsukada, and Y. Imai. 1995. Attachment and growth of fibroblast cells on silk fibroin. Biochem. Biophys. Res. Commun. 208: 511516.
14 Gotoh, K., H. Izumi, T. Kanamoto, Y. Tamada, and H. Nakashima. 2000. Sulfated fibroin, a novel sulfated peptide derived from silk, inhibits human immunodeficiency virus replication in vitro. Biosci. Biotechnol. Biochem. 64: 16641670.
15 Meisel, H. 1997. Biochemical properties of bioactive peptides derived from milk proteins: potential nutraceuticals for food and pharmaceutical applications. Livestock Prod. Sci. 50: 125-138.
16 Minoura, N., M. Tsukada, and M. Nagura. 1990. Physicochemical properties of silk fibroin membrane as a biomaterial. Biomaterials 11: 430--434.
17 Nishioka, K., A Constantopoulos, P. S. Satoh, and V. A Najjar. 1972. The characteristics, isolation and synthesis of the phagocytosis stimulating peptide tuftsin. Biochem. Biophys. Res. Commun. 47: 172-179.
18 Takano, R, K. Hirabayashi, and K. Chen. 1991. Preparation of soluble silk fibroin powder by hydrochloric acid hydrolysis. Sov. Phys. Crystallogr. 60: 358-362.
19 Kim, D. K., Y. H. Kim, K.. B. Kim, and Y. G. Chin. 2001. The changes of molecular weight and structure in the preparation process of silk fibroin powder. J. Kor. Fiber Soc. 38: 105-110.
20 Hanawa, T.,A Watanabe, T. Tsuchiya, R Ikoma, M. Hidaka, and M. Sugihara. 1995. New oral dosage form for elderly patients: preparation and characterization of silk fibroin gel. Chem. Pharm. Bull. 42: 282-288.
21 Gill, I., R Lopez-Fandino, X. Jorba, and E. N. Vulfson. 1996. Biologically active peptides and enzymatic approaches to their production. Enz. Microb. Techno!. 18: 162-183.
22 Hyun, C. K. and H. K. Shin. 2000. Utilization of bovine blood plasma proteins for the production of angiotensin I converting enzyme inhibitory peptides. Process Biochem. 36: 65-71.   DOI   ScienceOn
23 Nathan, C. 1992. Nitric Oxide as a secretory product of mammalian cells. FASEB J. 6: 3051-3064.
24 Luo, J., K. Chen, Q. Xu, and K. Hirabayashi. 1993. Study on foodization of fibroin and its functionality, pp. 73-87. In The collection of papers for the second international silk conference, China.
25 Stuehr, D. J., S. S. Gross, I. Sakuma, R. Levi, and C. F. Nathan. 1989. Activated murine macrophages secrete a metabolite of arginine with the bioactivity of endotheliumderived relaxing factor and the chemical reactivity of nitric oxide. J. Exp. Med. 169: 1011-1120.
26 Nathan, C. 1995. Natural resistance and nitric oxide. Cell 82: 873-876.
27 Schlimme, E. and H. Meisel. 1995. Bioactive peptides derived from milk proteins. Structural, physiologicaland analytical aspects. Die Nahrung 39: 1-20.
28 Nahm, J. H. and Y. S. Oh. 1995. A Study of pharmacological effect of silk fibroin. RDA J. Agric. Sci. 37: 145-157.