초록 ▼

○ 연구결과
- 자색고구마로부터 안토시아닌 물질의 대량 분리방법 확립(추출·농축)
- 추출 및 분리방법에 따른 지표물질 및 확인물질의 특성
- 자색고구마 추출박의 성분분석 및 알콜 발효조건 최적화
- 자색고구마 막걸리·식초의 알콜발효조건, 초산발효조건 확립 및 최적화
- 자색고구마 막럴리 및 자색고구마 식초의 대량생산공정 표준화
- 개발제품의 자가품질검사 기준·규격 확립, 시제품 제작 및 포장디자인 개발
- ROS 소거능, superoxide 소거능, 세포사멸능, GSH, 지질과산화 측정

○ 연구결과
- 자색고구마로부터 안토시아닌 물질의 대량 분리방법 확립(추출·농축)
- 추출 및 분리방법에 따른 지표물질 및 확인물질의 특성
- 자색고구마 추출박의 성분분석 및 알콜 발효조건 최적화
- 자색고구마 막걸리·식초의 알콜발효조건, 초산발효조건 확립 및 최적화
- 자색고구마 막럴리 및 자색고구마 식초의 대량생산공정 표준화
- 개발제품의 자가품질검사 기준·규격 확립, 시제품 제작 및 포장디자인 개발
- ROS 소거능, superoxide 소거능, 세포사멸능, GSH, 지질과산화 측정
- DNA 손상, 세포사멸 관련 Fas/FasL에 대한 자색고구마 안토시아닌의 보호효과 조사
- 자색고구마 안토시아닌의 항산화 효소계 및 산화적 손상 물질에 대한 영향 조사(in vivo)

Abstract ▼

This study was conducted to investigate the effect of the antioxidative activity and develop the processed food from anthocyanin of purple sweet potato(PSP).
The extraction yield of anthocyanin from purple-sweet potato was investigated by
measuring TOD(Total Optical Density) and showed approxi

This study was conducted to investigate the effect of the antioxidative activity and develop the processed food from anthocyanin of purple sweet potato(PSP).
The extraction yield of anthocyanin from purple-sweet potato was investigated by
measuring TOD(Total Optical Density) and showed approximately 90.7%, which comparing 6.35±0.50 in TOD with 7 as maximal optical density of anthocyanin at λ max from purple-sweet potato.
Anthocyanins from purple-sweet potato are greatly stable pigments including a structure of peonidin with acylated forms of ferulic acid and caffeic acid and, the pigments of peonidin, which occupy the highest ratio as peonidin-3-diglucoside-5-glucoside, are water-soluble. So we determined peonidins as the functional substance.
To establish the database for the functional marker, we concentrated peonidin-3-diglucoside-5-glucoside from ethanol extracts of purple-sweet potato and established the manufacturing standards according to Law for Health Functional Foods.
Including purple-sweet potato, various plants contain anthocyanin pigments, which is the main obstacle to generalize an analysis method for a specific kind. In this study, we discussed different analysis methods, reported recently, and finally chose A.O.A.C. analysis and HPLC analysis as the most effective method.
The antioxidant activities of anthocyanin extract in vitro, conducted by co-researching team for this study, exhibited non-cytotoxicity at 800ug/ml and antioxidant activities experiments with ROS scavengmg activity, superoxide scavenging activity, lipid peroxidation and DPPH free radical scavenging activity showed significant results dose-dependantly from 50ug/ml. This results were applied to database for the dose of intake in vivo.
The nutritional properties of purple-sweet potato residues were investigated and exhibited 330kcal/100g, 81.7g/100g, 5.4g/100g, 4.4g/100g and 22.7g/100g in calorie, carbohydrate, crude protein, sugar and dietary fiber, respectively.
The pH and acidity during the rice wine fermentation using PSP and the residue of PSP extracts decreased significantly to 4.51 and 0.3% in early phase and showed 3.85 and 4.2% after 15 days, respectively. The sugar content was immediately 4.8 brix after inoculation. As the fermentation progressed, the sugar content tended to increase and showed 6.2 brix after 15 days. The alcohol content during fermentation was 4.3% on the 1st day. After two days, the alcohol increased sharply to 6.0%. There was not significant difference with reaching to 6.5% after 15 days.
To isolate wild acetobacter from PSP, PSP vinegar starter fermented during 15 days was inoculated to culture media. First, 6 strains forming colonies were obtained, which were purified and inoculated to YPM broth for 48 hours at 30℃.
As high-productive acetobacter, WT-01, WT-02, WT-03 were selected among the isolated 6 strains by measuring clear zones in paper disc. The strains showed great acetic acid productivity forming over 2.0mm clear zone. In particular, WT-01 showed exclusive activity with 4.8mm clear zone.
The diverse zones surrounding the discs were analyzed in strains(WT-01, WT-02, WT-03) obtained by paper disc method. WT-01, WT-02 showed 1.8 mm and 1.0 mm in growth zone, 3.4 mm and 2.4 mm in clean zone and, 0.1 mm and 0.2 mm in effect zone, respectively. WT-03 showed great fermentablity with 1.2 mm in growth zone, 1.8 mm in clear zone, 0.1 mm in effect zone.
To investigate fermentability by the substrate consumption and chemical change, the changes in pH, total acidity, sugar content, alcohol content and CFU were observed by standard analysis for 15 days after inoculating the strains to the PSP fermentation extracts. Sugar content tended to increase continuously to 6 ± 0.5%. Alcohol content increased to 2 - 3% after inoculation, but was adversed in the late phase of fermentation. Also, pH tended to decrease continuously during fermentation and there is no significant difference among extracts fermented for 3 days. In total acidity, the content of acetic acid multiplied approximately 4 times compared to the content before inoculation, which produced over 7.6% acidity. In particular, the PSP fermentation extract by WT-01 showed the highest acidity producing 9.0% acidity. CFU for acetobacter increased sharply from 7th day and detected over 1.0 x 10⁵ CFU/㎖ after 15 days.
The comparative experiment for fermentability between two different fermentation substrates by WT-01 and KCTC 1010, showed 5.7 ± 0.2 and 3.84 ± 0.06 in brix and pH, which indicated no significant difference before and after inoculating the acetobacter.
Alcohol content increased to 2 - 3% after inoculating the acetobacter, but reduced in late phase of fermentation.
Here, we developed the functional food materials with cytoprotective effect and antioxidant activity against oxidative stress by using anthocyanin from PSP and identified its effect on animal model in vivo. also, we developed the functional rice wine(Makgeolli) and vinegar to establish scientific basis to promote antioxidative functional food using PSP anthocyanin by identifying the mechanism of the natural physiological antioxdant.
In the present work, we investigated the protective ettects of anthocyanin fraction (AF) from purple sweeet potato on tert-butyl hydroperoxide (t-BHP)-induced hepatotoxicity in HepG2 cell line and in rat liver. The result showed that the oral pretreatment of AF before t-BHP treatment significantly lowered the serum levels of the hepatic enzyme markers (ALT and AST) and reduced oxidative stress of the liver by evaluation of malondialdehyde and glutathione. Histopathological evaluation of the also revealed that AF reduced the incidence of liver lesions. The in vitro resultshowed that AF significantly reduced t-BHP-induced oxidative injury, as determined by cell cytotoxicity, intracellular glutathione content, lipid peroxidation, reactive oxygen species (ROS) levels, and caspases activation. Also, AF up-regulated antioxidant enzymes including heme oxygenase-1 (HO-1), NAD(P)H:quinone reductase, and glutathione S-transferase. Moreover, AF inNrf2 nuclear translocation and Akt and ERK1/2 activation, pathways that areinvolved in inducing Nrf2 nuclear translocation.
We examined whether an anthocyanin fraction (AF) from purple sweet potato may prevent dimethylnitrosamine (DMN)-induced liver injury by inducing antioxidants via nuclear erythroid 2-related factor 2 (Nrf2) pathways and by reducing inflammation. Treatment with AF attenuated the DMN-tnduced increased serum alanine aminotransferase and aspartate aminotransferase activities. It also prevented the formation of hepatic malondiadehyde and the depletion of glutathione and maintained normal glutathione-S-transferase (GST) activity in the livers of DMN-intoxicated rats. Furthermore, AF increased the expression of Nrf2, NADPH:quinine oxidoreductase-1, heme oxygenase-1. and GSTα, which were reduced by DMN, and decreased the expression of cyclooxygenase-2 and inducible nitric oxide synthase. An increase in the nuclear translocation of nuclear factor kappa B (NF-κB) was observed in DMN-induced oiver injury group, but AF inhibited this translocation.
In current study, we investigated the protective effects of the anthocyanin fraction (AF) obtained from the purple-fleshed sweet potato on hepatic fibrosis induced by dimethylnitrosamine (DMN) administration in rats. Treatment with DMN for 4 weeks produced marked liver fibrosis as assessed by increased serum alanine aminotransferase and aspartate aminotransferase activity and hepatic collagen content. These increases were inhibited by treatment with AF prior to the administration of DMN. In addition, AF inhibited DMN-induced reductions in rat body and liver weights in a dose-dependent manner. Histopathological evaluation of the rat livers revealed that AF reduced the incidence of hepatic fibrosis lesions and inhibited DMN-induced increases in –smooth muscle actin (α-SMA) and collagen type I and III expression levels. AF also decreased DMN-induced expression levels platelet-derived growth factor receptors-beta, tumor necrosis factor-alpha and transforming growth factor-beta.
We also investigated the anti-proliferative effects of an anthocyanin fraction (AF), isolated from the purple-fleshed sweet potato, on platelet-derived growth factor (PDGF)-BB-dependent signaling pathways in HSC-T6 cells. HSC proliferation plays a
pivotal role in liver fibrogenesis. The AF suppressed HSC activation, including PDGF-induced proliferation and α-smooth muscle actin (αSMA) expression. Additionally, AFinhibited PDGF-BB-induced Akt and ERKl/2 phosphorylation. AF inhibited the phosphorylation level of PDGF receptorb (PDGFR-b) following PDGF-BB stimulation, providing a mechanism for the inhibition of AF-mediated kinase.
Here, we developed the functional dietary materials with cytoprotective effect and antioxidant activity against oxidative stress by using anthocyanin from PSP and identified its effect on animal model in vivo. Also, we developed the functional rice wine(Makgeolli) and vinegar to establish scientific basis to promote antioxidative functional food using PSP anthocyanin by identifying the mechanism of the natural physiological antioxidant.

목차 Contents

• 표지 ... 1
• 요약문 ... 3
• 목차 ... 23
• 제 1 장 연구개발과제의 개요 ... 27
• 1. 연구개발의 목적 ... 27
• 2. 연구개발의 필요성 ... 27
• 3. 연구개발의 내용 및 범위 ... 29
• 제 2 장 국내외 기술개발 현황 ... 32
• 1. 국외 현황 ... 32
• 2. 국내 현황 및 문제점 ... 32
• 제 3 장 연구개발 수행내용 및 결과 ... 34
• 제1절 자색고구마 및 추출박을 이용한 안토시아닌 함유 가공식품의 개발 및 상품화 ... 34
• 1. 자색고구마 안토시아닌 색소의 기준 및 규격 확립 및 자색고구마 추출박을 이용한 자색 고구마 막걸리의 개발 ... 34
• 2. 자색고구마 추출박을 활용한 자색식초의 개발 ... 58
• 3. 자색고구마 추출박을 활용한 자색고구마 막걸리 및 자색식초의 상품화 ... 77
• 제2절 자색고구마 안토시아닌의 항산화 기능성 검증 및 기능성 성분-원료의 in vitro, in vivo 기능성 조사 ... 92
• 1. 자색고구마 안토시아닌 색소의 항산화 기능성 검증(in vitro 항산화 기능성 평가) ... 92
• 2. in vivo 실험동물에서 tert-butyl hydroperoxide (t-BHP)에 의한 산화적 간 손상에 대한 자색고구마 안토시아닌의 간 보호 효과 조사 ... 112
• 3. in vivo 실험동물에서 dimethylnitrosamine (DMN)에 의한 산화적 간손상에 대한 자색고구마 안토시아닌의 간 보호 효과 조사 ... 118
• 4. 자색고구마 안토시아닌의 강성상세포( hepatic stellate cells) 증식 억제효과 및 작용기전 규명 ... 127
• 제 4 장 목표달성도 및 관련분야에의 기여도 ... 134
• 1. 목표달성도 ... 134
• 2. 관련분야 기여도 ... 137
• 제 5 장 연구개발 성과 및 성과활용 계획 ... 138
• 1. 연구개발 성과 및 활용계획 ... 138
• 제 6 장 연구개발과정에서 수집한 해외과학기술정보 ... 144
• 제 7 장 참고문헌 ... 145
• 부 록 ... 152
• 끝페이지 ... 206