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해마 (Hippocampus abdominalis) 효소가수분해물의 항산화 활성 및 생리활성 펩타이드의 분리

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Alternative Title
Antioxidant activity of enzymatic hydrolysates from seahorse (Hippocampus abdominalis) and isolation of its bioactive peptides
Abstract
Seahorse, Hippocampus abdominalis (H. abdominalis) known as a marine teleost fish, has been used as one of the most famous and expensive materials of traditional Chinese medicine due to its beneficial biological activities. However, its use has been limited by its high cost despite its beneficial biological activities and there are few reports on its biological activity and active components. Therefore, we investigated the antioxidant activity of H. abdominalis and its active compounds. First of all, to improve its extraction yields, we prepared the water-enzymatic hydrolysates by performing the water and enzymatic extraction techniques with the seven food grade enzymes (Trypsin, Pepsin, Neutrase, Protamex, Alcalase, Kojizyme, and Flavourzyme) in consecutive order. The enzymatic extracts showed the higher extraction yield than that of the only water extract. Among them, the water-Pepsin hydrolysate showed the higher DPPH, hydroxyl and alkyl radicals, and hydrogen peroxide scavenging activities. Also, it protected against oxidative stress caused by the treatment of AAPH. To identity its active components, we fractionated the > 5 KDa and < 5 KDa molecular weight fractions from the Water-Pepsin hydrolysates and confirmed the superior antioxidant activity of < 5 kDa frction. The < 5 KDa fraction, a low molecular weight fraction was applied to Sephadex G-25 gel filtration chromatography and we identified that the isolated most low molecular weight fraction led to the highest alkyl radical scavenging activity. Results of examining amino acid composition of the active peptide, it was possible to confirm the presence of peptides consisting with mainly many tryptophan and glycine. In conclusion, in this study suggests that the seahorse water-enzymatic hydrolysates prepared by the useful water-extraction techniques has the antioxidant activity and might be an useful resource of marine origin as a novel material for healthy functional foods and cosmetics.
Author(s)
신병옥
Issued Date
2015
Awarded Date
2016. 2
Type
Dissertation
URI
http://dcoll.jejunu.ac.kr/jsp/common/DcLoOrgPer.jsp?sItemId=000000007592
Alternative Author(s)
Shin, Byeung-Ok
Department
대학원 해양생명과학과
Advisor
전유진
Table Of Contents
1. 서론 1
2. 재료 및 방법 6
2. 1. 실험재료 6
2. 2. 시약 6
2. 3. 해마의 일반성분 분석 7
2. 4. 단백질 효소가수분해물의 제조 7
2. 5. 단백질 효소가수분해물의 수율 측정 11
2. 6. 단백질 효소가수분해물의 총단백질 측정 11
2. 7. 자유 라디칼 (free radical) 소거활성의 측정 11
2. 7. 1. DPPH radical 12
2. 7. 2. Hydroxyl radical 12
2. 7. 3. Alkyl radical 13
2. 7. 4. Hydrogen peroxide 13
2. 8. Vero cell 에서의 항산화 활성 측정 14
2. 8. 1. Cell culture 14
2. 8. 2. Cytotoxicity 15
2. 8. 3. Total ROS 15
2. 8. 4. Cell viability 16
2. 9. Zebrafish embryo 에서의 항산화 활성 측정 17
2. 10. 생리활성 펩타이드의 분리 18
2. 10. 1. Ultrafiltration 18
2. 10. 2. Gel filtration chromatography 18
2. 11. Amino Acid composition analysis 19
3. 결과 21
3. 1. 해마의 일반성분 21
3. 2. 단백질 효소가수분해물의 수율 21
3. 3. 단백질 효소가수분해물의 총단백질 24
3. 4. 자유 라디칼 (free radical) 소거활성의 평가 26
3. 4. 1. DPPH radical 26
3. 4. 2. Hydroxyl radical 29
3. 4. 3. Alkyl radical 32
3. 4. 4. Hydrogen peroxide 35
3. 5. Vero cell 에서의 항산화 활성 평가 38
3. 5. 1. Cytotoxicity 38
3. 5. 2. Total ROS 40
3. 5. 3. Cell viability 42
3. 6. Ultrafiltration 44
3. 6. 1. Alkyl radical 소거활성의 평가 44
3. 6. 2. Vero cell 에서의 항산화 활성 평가 46
3. 6. 2. 1. Cytotoxicity 46
3. 6. 2. 2. Total ROS 46
3. 6. 2. 3. Cell viability 49
3. 6. 3. Zebrafish embryo 에서의 항산화 활성 평가 51
3. 7. Sephadex G-25 gel filtration chromatogram 53
3. 7. 1. Alkyl radical 소거활성의 평가 53
3. 7. 2. Amino Acid composition 56
4. 고찰 60
5. 참고문헌 64
6. 감사의 글 68
Degree
Master
Publisher
제주대학교 대학원
Citation
신병옥. (2015). 해마 (Hippocampus abdominalis) 효소가수분해물의 항산화 활성 및 생리활성 펩타이드의 분리
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General Graduate School > Marine Life Sciences
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