발효누에분말의 생리활성

국문 초록 (Abstract)

본 연구에서는 누에 단백질을 미생물발효에 의해 생리활성물질로 개발하기 위한 기초 연구의 일환으로 단백질 분해능이 높은 Bacillus 및 Lactobacillus 균주로 동결건조 및 열풍건조 누에분말을 발효시켜 항산화 활성, 혈전용해 활성, tyrosinase 저해 활성, 단백질 패턴 비교 등을 비교 검토하였다. Native-PAGE 및 SDS-PAGE로 누에 단백질을 분석 한 결과 동결건조 누에분말에서 많은 종류의 밴드가 관찰되었고, B. subtilis 발효에 의해서 단백질 패턴과 양적인 차이가 관찰되었다. 열풍 및 동결건조 누에분말의 주요 미네랄은 칼륨, 칼슘, 마그네슘 및 아연 이였으며, 주요 아미노산은 glycine, alanine, glutamic acid, aspartic acid 및 serine 이였고, 주요 지방산은 linolenic acid, oleic acid 및 linoleic acid 이였다. 혈전용해 활성은 동결 및 열풍건조 누에분말 모두 B. subtilis 균주 5% 접종에서 가장 높았으며, 동결건조 발효누에서 보다 높은 혈전용해 활성이 나타났다. Tyrosinase 저해 효과는 열풍건조 누에분말 보다 동결건조 누에분말의 물 및 70% 메탄올 추출물에서 높았다. DPPH free-radical 소거능은 동결건조 누에분말 보다 열풍건조 누에분말에서 조금 높게 나왔으며, L. hilgardii 균주에 의한 발효누에가 B. subtilis 균주에 의한 발효 누에보다 항산화 활성이 약간 더 높은 것으로 관찰되었으나, 모든 시료에서 시판 항산화제인 BHT 보다는 낮았다. 이상의 실험결과에서 누에 단백질을 생리활성물질로 개발하기 위한 기초 자료를 얻기 위하여 단백질 분해능이 높은 B. subtilis 균주로 발효시킨 발효누에에서 단백질 함량 증가, 전기영동 단백질 패턴 변화, 혈전 용해 활성 및 항산화 활성 증가와 같은 생리활성작용이 강화됨으로써 건강기능식품 소재 개발 가능성이 확인되었다.

다국어 초록 (Multilingual Abstract)

The comparative effects of the fibrinolytic, and tyrosinase inhibition activities and electrophoretical protein patterns with freeze-drying silkworm powder (FDSW), heating-drying silkworm powder (HDSW) and fermented silkworm powder by Bacillus subtilis or Lactobacillus hilgardii were investigated. When total protein patterns of FDSW, HDSW, both fermented SW, were analyzed by native- and SDS-polyacrylamide gel electrophoresis (PAGE), there were slightly varietal differences in electrophoretical protein patterns. Major minerals of FDSW and HDSW were K, Ca, Mg, and Zn. Major compositional amino acids of FDSW and HDSW were glycine, alanine, glutamic acid, aspartic acid, and serine. Major fatty acids of FDSW and HDSW were linolenic acid, oleic acid, and palmitic acid. Fibriolytic activity was the highest in the fermented FDSW by 5% B. subtilis among the various samples. Tyrosinase inhibition activity was higher in the water and 70% methanolic extract of FDSW than in HDSW. DPPH radical scavenging activity was slightly stronger in HDSW than in FDSW. In addition, DPPH radical scavenging activity was higher in FDSW or HDSW fermented by L. hilgardii than that fermented by B. subtilis, however, all samples exhibited a relatively low activity compared to the butylated hydroxytoluene (BHT). These results may provide the basic data to understand the biological activities of fermented SW.

목차 (Table of Contents)

• 서론
• 재료 및 방법
• 결과 및 고찰
• 요약
• 감사의 글
• References

참고문헌 (Reference)

1 김익조, "혈전용해효소함유 기능성 청국장제조에 관한 연구" 한국생명과학회 12 (12): 357-362, 2002

2 오혜숙, "팥과 녹두의 이소플라빈 함량과 항산화 및 혈전용해 활성" 한국식품조리과학회 19 (19): 263-270, 2003

3 류희영, "자생곤충으로부터 트롬빈 저해물질의 탐색 및 DPPH radical 소거활성" 한국생명과학회 18 (18): 363-368, 2008

4 이상한, "약용곤충추출물 라이브러리를 이용한 항산화 활성의 초고속 검색" 한국식품저장유통학회 12 (12): 482-488, 2005

5 조영수, "된장 유래 혈전분해효소 생산균주의 분리 및 최적 효소생산 조건 탐색" 한국식품저장유통학회 12 (12): 643-649, 2005

6 양선아, "단삼 메탄올 추출물의 항혈전 및 항산화 효과" 한국식품과학회 39 (39): 83-87, 2007

7 정용진, "녹두의 가수분해조건에 따른 특성 변화" 한국식품저장유통학회 13 (13): 563-568, 2006

8 Weber,K.and M.Osborn, "The reliability of molecular weight determination by sodium dodesyl sulphate-polyacrylamide gel electrophoresis" 244 : 4406-4412, 1969

9 Astrup, T, "The fibrin plate method for estimating fibrinolytic activity" 40 : 346-351, 1991

10 Lee, S., "Synthesis and melanin biosynthesis inhibitory activity of (+/-)-terrein produced by Penicillium sp. 20135" 15 : 471-473, 2005

11 Kato, N., "Silk protein, sericin, inhibits lipid peroxidation and tyrosinase activity" 62 : 145-147, 1998

12 Shiomi, S., "Significance of peptidoglycan in patients with chronic liver diseases" 47 : 32-37, 1998

13 Jung, S. W., "Screening of tyrosinase inhibitor from plants" 27 : 891-896, 1995

14 Kim, K. T., "Protective effect of steamed American ginseng (Panax quinquefolius L.) on V79-4 cells induced by oxidative stress" 111 : 443-450, 2007

15 Lee, J. H., "Protective effect of selected amino acids and food extracts on ethanoltoxicity determent in ratee and" 31 : 802-808, 1999

16 Garces, R, "One-step lipid extraction and fatty acid methyl esters preparation from fresh plant tissues" 211 : 139-143, 1993

17 AOAC, "Official methods of analysis. 17th eds" Association of official analytical chemists 1997

18 Masamoto, Y., "Mushroom tyrosinase inhibitory activity of esculetin isolated from seeds of Euphorbia lathyris L" 67 : 631-634, 2003

19 Duncan,D.B, "Multiple range and multiple F test" 1 : 1-42, 1959

20 Yun, E. Y., "Molecular cloning and expression of the novel attacin-like antibacterial protein gene isolated from the Bombyx mori" 36 : 331-340, 1997

21 Cabanes, J., "Kojic acid, a cosmetic skin whitening agent, is a slow binding inhibitor of catecholase activity of tyrosinase" 46 : 982-985, 1994

22 Ok, M., "Isolation and characterization of Bacillus sp. WRD-2 extracellular protease from soil" 44 : 246-250, 2001

23 Arena, M. E, "Influence of ethanol and low pH on arginine and citrulline metabolism in lactic acid bacteria from wine" 156 : 858-864, 2005

24 Stuart, M. R., "Influence of carbohydrate starvation and arginine on culturability and amino acid utilization of Lactococcus lactis sub sp. lactis" 65 : 665-673, 1999

25 Kang, K. S., "Increase in the free radical scavenging activity of ginseng by heat-processing" 29 : 750-754, 2003

26 Yin, M., "Glycine accelerates recovery from alcohol-induced liver injury" 286 : 1014-1019, 1998

27 Kang, P. D., "Electrophoretic patterns of hemolymph proteins of varieties with long and short life span in the silkworm Bombyx mori L" 41 : 1-8, 1999

28 Sohn, H. D, "Effects of juvenile hormone analogue on changes of protein and amino acids in haemolymph of the silkworm, Bombyx mori" 28 : 37-42, 1986

29 Ryu, K. S., "Effects of Bombyx mori L. extracts on carbon tetrachloride-induced hepatotoxicity in mice" 9 : 375-381, 1999

30 Cho, C. H., "Effect of temperature, time and pH on the extraction of protein in a chrysalis of silkworm" 4 : 65-68, 1989

31 Chung, S. H., "Effect of silkworm extract on intestinal α-glucosidase activity in mice administrated with a high carbohydrate-containing diet" 39 : 86-92, 1997

32 Kang, G. D., "Effect of silk fibroin on the protection of alcoholic hepatotoxicity in the liver of alcohol preference mouse" 2 : 15-18, 2001

33 Seong, S. I., "Effect of metamorphois on the major hemolymph proteins of the silkworm" 2 : 91-104, 1985

34 김용순, "Effect of Silkworm (Bombyx mori) Excrement Powder on the Alcoholic Hepatotoxicity in Rats" 한국생명과학회 18 (18): 1342-1347, 2008

35 Davis, B. J, "Disc electrophoresis-II. Method and application to human serum proteins. In gel electrophoresis" Acad. Sci. 121-404, 1964

36 Yukio Suzuki, "Arginyl-fructosyl-glucose and Arginyl-fructose, Compounds Related to Broming Reaction in the Model System of Steaming and Heat-drying Processes for the Preparation of Red Ginseng" 고삼학회 28 (28): 143-148, 2004

37 Tonon, T, "Arginine metabolism by wine Lactobacillus isolated from wine" 19 : 451-461, 2002

38 Cha, J. Y., "Antioxidative activities and contents of polyphenolic compound of Cudrania tricuspidata" 28 : 1310-1315, 1999

39 Nicoli, M. C., "Antioxidant properties of coffee brews in relation to the roasting degree" 30 : 292-297, 1997

40 Blois, M. S, "Antioxidant determination by the use of a stable free radical" 26 : 1199-1204, 1958

41 Sumi, H., "A novel fibrinolytic enzyme (Nattokinase) in the vegetable che in Natto: a typical and popular soybean food in the Japanese diet" 43 : 1110-1111, 1978