보고서 정보
주관연구기관 |
산동농협협동조합 |
보고서유형 | 최종보고서 |
발행국가 | 대한민국 |
언어 |
한국어
|
발행년월 | 2013-07 |
과제시작연도 |
2012 |
주관부처 |
농림축산식품부 Ministry of Agriculture, Food and Rural Affairs(MAFRA) |
등록번호 |
TRKO201400000121 |
과제고유번호 |
1545004177 |
사업명 |
고부가가치식품기술개발 |
DB 구축일자 |
2014-05-07
|
초록
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○ 연구결과
맥문동의 증숙의 최적화를 통해서 증숙 맥문동(홍문동)을 제조하였고,맥문동 및 홍문동의 기능성 증진을 위해 항산화능과 ACE저해능이 우수한 균주를 활용하여 발효 맥문동 및 발효 홍문동을 제조하였다. 제조된 홍문동,발효 맥문동 및 발효 홍문동의 생리활성 성분을 탐색하고 증숙 및 발효에 따른 생리활 성성분의 변화를 분석하였으며,또한 항당뇨 및 인지개선과 관련된 효능을 in vitro와 in vivo실험을 통해서 검증하였으며,관련제품의 허가를 위한 독성시험,효능실험 및 식약청 허가자료를 확보하여 제품인증을 추진하였다.그리
○ 연구결과
맥문동의 증숙의 최적화를 통해서 증숙 맥문동(홍문동)을 제조하였고,맥문동 및 홍문동의 기능성 증진을 위해 항산화능과 ACE저해능이 우수한 균주를 활용하여 발효 맥문동 및 발효 홍문동을 제조하였다. 제조된 홍문동,발효 맥문동 및 발효 홍문동의 생리활성 성분을 탐색하고 증숙 및 발효에 따른 생리활 성성분의 변화를 분석하였으며,또한 항당뇨 및 인지개선과 관련된 효능을 in vitro와 in vivo실험을 통해서 검증하였으며,관련제품의 허가를 위한 독성시험,효능실험 및 식약청 허가자료를 확보하여 제품인증을 추진하였다.그리고,제조된 홍문동,발효 맥문동 및 발효 홍문동을 이용한 차음료,액기스 형태의 시제품을 제작하여 관능검사 및 소비자 조사를 수행하였으며,제품의 생산을 위한 제품 디자인, 성분분석 및 품질관리와 관련된 시험성적서 등을 완료하였다.
Abstract
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IV. Results of research and development
We tried to optimize the steaming and drying process of Liriopis tuber which is one of the ways of food processing, thereby increasing the content of bioactive components in Liriopis tuber. Changes of general components, crude saponin, total phenolic compou
IV. Results of research and development
We tried to optimize the steaming and drying process of Liriopis tuber which is one of the ways of food processing, thereby increasing the content of bioactive components in Liriopis tuber. Changes of general components, crude saponin, total phenolic compounds, flavonoid compounds, and antioxidative activities were analyzed according to steaming times and and steaming frequencies. From the results, the condition for steaming during 15 hours and drying for 72 hours at 70 °C was fhe best condition to maximize the biologically active compounds. During the steaming and drying process, newly forming toxic substances such as benzopyrene was not detected.
To analyze the bioactive components of steamed Liriopis tuber (red Liriopis tuber), the methanol extract and the butanol extract were prepared. From these extracts, phenolic compound, saponin, amino sugars, and sugar components were analyzed. Pure compound produced during steaming process was purified by Prep HPLC and recycling HPLC. In addition, large amounts of fractions was prepared by Prep HPLC and fheir anti-diabetic effect on insulin secretion was confirmed, showing browning reaction produced during the steaming process to be related with insulin secretion.
From the analysis of fermented Liriopis tuber and fermented red Liriopis tuber, similar bioactive compounds found in red Liriopis tuber were observed, which is due to the high temperature and high pressure during the extraction process. Especially color of the extracts, contents of HMF and amino sugar were increased in both cases. Spicatoside A, one of the important saponin compounds in Liriopis tuber was decreased, but diosgenin and ruscogenin were newly produced in fermented Liriopis tuber and fermented red Liriopis tuber.
Fermentation by microorganisms could enhance biological activity, as well as produce de novo biologically active compounds. For the fermentation of Liriopis tuber, microorganisms (Lactobacillus 74 strains, Bacillus 50 strains) were screened from fhe Korea traditional food, and the surface of various plants. The biological activity of these strains was confirmed by antioxidant activity, anti-hypertensive activity (ACE inhibition activity), antimicrobial activity, acid and bile acid resistance, enzymes activity toward cellular components.
Two strains having antioxidative activity and ACE inhibitory activity were selected for further fermentation. Wifh two strain, fermentation condition was optimized to maximize the antioxidative activity and ACE inhibitory activity. Also, effects of Liriopis tuber extraction condition, growth rate of selected strains, and biological activities of culture broth were analyzed. Based on the fermentation pattern, two strain showed the reduction of pH and the production of titrable acid fhrough the fermentation.
Among screened strains, Lactobacillus brevis DD1 was finally chosen according to DPPH radical scavenging activity and ACE inhibitory activity for fhe fermentation of Liriopis tuber and red Liriopis tuber. Fermentation by this strain was monitored and optimized based on incubation temperature, initial pH and amounts of initial inoculum. In addition, various cultures parameters (titratable acidity, cell growth, too, total sugar and reducing sugar) and physiological activity (DPPH radical scavenging activity, ACE inhibitory activity, polyphenolic compounds and flavonoids) were investigated during the fermentation. From these results, biological activities were increased during the fermentation and ACE inhibitory activity was doubled by the fermentation process.
Optimal fermentation condition of hot water extract for the best ACE inhibitory activity was pH 4.5, 35°C, and 4% (v/v) initial inoculum, and optimal fermentation condition of high temperature and high pressure water extract for the best ACE inhibitory activity was pH 5.0, 30°C, and 6% (v Iv) initial inoculum, respectively.
Effects of steaming conditions on the physiological activity of the cells were investigated in terms of anti-diabetics and memorial improvement. As a result, culture conditions of the B35 neuronal cell line, PC12 pheochromocytoma cell line, and INS cell lines were established. Toxicity and optimal concentration for insulin secretion was measured usmg various red Liriopis tuber extracts. In INS cell, highest insulin secretion was observed in 3 hour and 9 times steamed and dried Liriopis tuber extracts. Also, insulin receptor signaling pathway was investigated. For memorial improvement by B35 neuronal cell, 9 hour and 7 times steamed and dried Liriopis tuber extracts showed highest NGF secretion activity. From these results, steaming and drying process of Liriopis tuber affects in insulin secretion and NGF secretion activity.
From in vitro results, the insulin secretion ability and glucose receptor signaling pathway was confirmed in an animal model of for type I diabetes. The following results were obtained: i) insulin secretion was induced in red Liriopis tuber extract-treated OLETF rats, although there were no significant differences in body weight, glucose tolerance test and glucose concentration; ii) fhe red Liriopis tuber extract-treated OLETF rats showed a significant increase in adiponectin concentration but the concentration of triglyceride and LDL decreased compared to the vehicle-treated rats; iii) although the abdominal fat mass and adipocyte size did not change with red Liriopis tuber extract treatment, expression of the adipocyte marker genes and β-oxidation genes in fat tissue was recovered to the level of the LETO rats; These results suggest that red Liriopis tuber extract may stimulate insulin secretion and a decrease in lipid in serum, and may also suppress fatty liver formation through the regulation of fatty acid oxidation. The data presented here highlight the possibility that red Liriopis tuber extract can be considered a candidate for the prevention or alleviation of obesity-related diseases. To examine fhe therapeutic effects of Red LP (RLP) manufactured by steaming process on neurodegenerative disorders, significant alteration of the key factors influencing Alzheimer's Disease (AD) was detected in NSE/hAPPsw transgenic (Tg) rmce after red Liriopis tuber extract treatment. The concentration of nerve growth factor (NGF) in serum increased in red Liriopis tuber extract-treated NSE/hAPPsw Tg mice compared with vehicle-treated Tg mice. Further, analysis of y-secretase components showed that Aβ-42 peptide expression was downregulated. Overall, fhese results suggest that red Liriopis tuber extract can help relieve neurodegenerative diseases, especially AD, through upregulation of NGF secretion ability, activation of NGF signaling pathway, downregulation of Aβ-42 peptide deposition, and alteration of y-secretase components.
The toxicity tests and efficacy test were investigated for approval of fhe final product and the data to promote the product certification by KFDA approval. Investigation of GLP protocol and single dose toxicity test was conducted to analyze the effects of red Liriopis tuber extract on body weight, hehepatotoxicity, nephrotoxicity with renal function test, which showed that red Liriopis tuber extract has no effects. As a non-clinical trial stage, the pharmacological effects and the mechanism of action as well as target organs to assess the effects of the undesirable pharmacological action were evaluated and red Liriapis tuber extract did not cause a toxic effect and showed relief of fhe immune response induced by P A. In addition, the pharmacokinetic study showed that red Liriopis tuber extract improved the constipation induced by loperamide.
To establish optimum conditions of extraction process, the extraction yield and antioxidative activity were analyzed on the hot water extract and ethanol extract. Hot water extract showed highest extraction yield with the high antioxidative activity. From these results, development of beverage products is suitable item for the industrial production and beverage product formulation and manufacturing conditions has been established. In addition, product name and desgine were fabricated and nutrient value was analyzed for the manufacturing beverage product. Sensory evaluation and cosurner's acceptance was investigated and these results would be adapted in final products.
For fhe development of various fermented Liriopis tuber and fermented red Liriopis tuber products, lactic acid bacteria selected from cooperative study was chosen, and its fermentation condition was established for the production of items. Product formulation and manufacturing conditions has been established and product desgine was selected. In addition, nutrient value was analyzed for the manufacturing the product. Sensory evaluation and cosurner's acceptance were investigated and these results would be also adapted in final products.
목차 Contents
- 표지 ... 1
- 제출문 ... 2
- 요약문 ... 3
- SUMMARY(영문요약문) ... 8
- CONTENTS ... 13
- 목차 ... 15
- 제 1 장 연구개발과제의 개요 ... 17
- 제 1 절 연구개발의 목적 ... 17
- 제 2 절 연구개발의 필요성 ... 18
- 제 3 절 연구개발의 범위 ... 21
- 제 2 장 국내외 기술개발 현황 ... 23
- 제 1절 국내 관련기술 현황 ... 23
- 제 2절 국내외 제품생산 및 시장현황 ... 25
- 제 3 장 연구개발수행 내용 및 결과 ... 28
- 제 1 절 제 1세부 홍문동의 기능성 제품 개발 및 산업화 ... 28
- 1. 홍문동의 기능성 제품 개발 ... 28
- 2. 발효 맥문동 기능성 제품 개발 ... 51
- 3. 발효 홍문동 기능성 제품 개발 ... 65
- 4. 홍문동 및 발효제품의 마케팅 및 홍보전략 수립 ... 77
- 제 2 절 제 1협동 맥문동을 이용한 홍문동의 제조 및 기능성 물질 분석 ... 78
- 1. 맥문동 증숙 조건의 확립 및 최적화 ... 78
- 2. 홍문동의 기능성 성분 분석 및 정제 ... 93
- 3. 홍문동의 기능성 성분 구조 분석 및 대량 분리 ... 120
- 4. 발효 맥문동의 기능성 성분 분석 ... 139
- 제 3 절 제 2협동 맥문동 및 홍문동 발효를 통한 기능성 증진 연구 ... 142
- 1. 발효균주의 분리,동정 및 기능성 조사 ... 142
- 2. 발효균주의 기능성 물질 생산조건 ... 158
- 3. 맥문동의 발효균주 증식효과 및 예비발효 ... 183
- 4. 맥문동 및 홍문동 발효 ... 190
- 제 4 절 제 3협동 홍문동의 생리활성 효능연구 ... 212
- 1. 생리활성 검증을 위한 췌장 세포주와 신경세포주의 확립 ... 212
- 2. In vitro 생리활성 효능평가 ... 212
- 3. 효능분석을 위한 모델동물 확립 ... 229
- 4. In vivo 생리활성 효능평가 ... 233
- 5. 홍문동의 독성시험 ... 253
- 6. 홍문동의 약리효능 평가 ... 262
- 제 4 장 목표달성도 및 관련분야에의 기여도 ... 287
- 제 1 절 연구개발의 달성도 및 기여도 ... 287
- 제 2 절 기술적 측면 ... 291
- 제 3 절 경제·산업적 측면 ... 291
- 제 4 절 사회·문화적 측면 ... 291
- 제 5 장 연구개발 성과 및 성과활용 계획 ... 292
- 제 1 절 특허 출원 ... 292
- 제 2 절 논문 ... 292
- 제 3 절 학술발표 ... 294
- 제 4 절 그 외 활용계획 ... 296
- 제 6 장 연구개발과정에서 수집한 해외과학기술정보 ... 297
- 제 1 절 맥문동 유래 신규 물질 정보 ... 297
- 제 2 절 증숙 공정관련 해외 기술정보 ... 297
- 제 2 절 효능평가를 위한 해외 질환모델 정보 ... 297
- 제 7 장 연구시설?장비 현황 ... 300
- 제 8 장 참고문헌 ... 301
- 연구개발보고서 초록 ... 306
- 끝페이지 ... 307
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