보고서 정보
주관연구기관 |
순천대학교 SunChon Natinal University |
보고서유형 | 최종보고서 |
발행국가 | 대한민국 |
언어 |
한국어
|
발행년월 | 2003-08 |
과제시작연도 |
2002 |
주관부처 |
농림부 Ministry of Agriculture and Forestry |
등록번호 |
TRKO201400023658 |
과제고유번호 |
1380002743 |
사업명 |
농림기술개발 |
DB 구축일자 |
2014-11-10
|
초록
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○ 연구결과
1) 산수유의 원료 전처리와 최적 효모와 초산균을 선정하여, 당 발효주, 누룩 발효곡주, 종국 발효곡주 및 발효음료를 개발하여 주요성분을 조사하였음.
2) 산수유의 항산화능, 항당뇨능 및 고지혈증 억제 효과를 in vitro 및 in vivo에서 검증하였음.
3) 산수유와 산수유 제품의 면역활성과 항암활성을 검증하고 항암활성 물질을 분리 확인하였음.
4) 기능성분을 다량 함유하는 추출물 제조법을 확립하고, 산수유를 이용한 음료 및 차의 제조법을 설정하였음.
5) 산수유의 떫은맛과 신맛 감소 방
○ 연구결과
1) 산수유의 원료 전처리와 최적 효모와 초산균을 선정하여, 당 발효주, 누룩 발효곡주, 종국 발효곡주 및 발효음료를 개발하여 주요성분을 조사하였음.
2) 산수유의 항산화능, 항당뇨능 및 고지혈증 억제 효과를 in vitro 및 in vivo에서 검증하였음.
3) 산수유와 산수유 제품의 면역활성과 항암활성을 검증하고 항암활성 물질을 분리 확인하였음.
4) 기능성분을 다량 함유하는 추출물 제조법을 확립하고, 산수유를 이용한 음료 및 차의 제조법을 설정하였음.
5) 산수유의 떫은맛과 신맛 감소 방법을 확립하였음
6) 산수유 추출물과 발효원액을 이용하여 액상 및 고상 제품의 대량 생산 제조공정을 확립하였음
7) 시제품의 제조 및 그 품질 특성을 조사하였음.
Abstract
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Ⅳ. Results and Suggestions for Application of the Results
1. Results
For the production of high quality wine with Corni fructus, the quality of raw Corni fructus was important in the point of color, low temperature natural drying and granule condition. Although fully ripened fruit was advantag
Ⅳ. Results and Suggestions for Application of the Results
1. Results
For the production of high quality wine with Corni fructus, the quality of raw Corni fructus was important in the point of color, low temperature natural drying and granule condition. Although fully ripened fruit was advantageous to immature one in color, fermentable sugar contents, sourness and astringency, the over-ripened fruit also can be used for the production of wine if it was not frozen on the tree.
The sourness and astringency of Corni fructus could be reduced by adjusting the initial ratio of sugar to Corni fructus to 6∼9. The sourness and astringency could also be controlled some by pretreatment of Corni fructus with semisolid Koji cake using Aspergillus oryge.
For the alcohol fermentation of Corni fructus, Saccharomyces sp. KOFRI 013 was selected as a proper yeast species, being judged with proliferation, production of ethanol and utilization of fermentable sugars. The ratio of Corni fructus to water was preferable in 5:1 or 7:1. As a fermentable sugar source, fructose and molasses were the proper ingredient at a concentration of 20%. Although alcohol production was slightly low, bee honey could also be used the fermentable sugar source as long as the taste is concerned.
The optimum temperature for the fermentation was found to be 20∼25℃.
The maximum alcohol concentration was 13.5% when fermented for 10∼20 days.
Fermentation at 5℃ obtained high scores in sensory evaluation, while fermentation at 15℃ produced sour taste and smell due to proliferation of acid producing bacteria.
To brew Corni fructus-Nuruk wine, the proper amount of Corni fructus
and Nuruck were 6∼10% and 4%, respectively. The alcohol production was 7.2∼7.6%, and the unfermented total sugar content was 7.02∼7.23%. Six days of main
fermentation at 25℃ followed by 60 days of ripening at 10℃ was found to be optimum for the production of Corni fructus-Nuruk wine.
A Koji wine was produced using Corni fructus. As a yellowish Koji, Aspergillus oryge was most efficient in digestion of polysaccharides to simple sugars which can be fermentable. Whitish Koji (Rhizopus oligosporus) and redish Koji had disadvantages of taking longer time to prepare Koji starter cake and low in amylase activity which results in low content of fermentable sugars, and consequently low in alcohol production. Whitish Koji produced less of bitter taste and sour taste, while overall acceptability was lower than yellowish Koji and redish Koji (Monascus anka). The adequate amount of Corni fructus added was 6∼10% considering the production of alcohol and overall acceptability. The optimum fermentation of mash was found to be 40∼60 days.
For the production of Corni fructus vinegar Acetobacter sp. KOFRI B-104 was selected as the starter bacteria. Optimum conditions for the fermentation were 1.5∼2% (w/v) of initial acidity, 7∼8% of initial alcohol concentration and incubation temperature of 30℃. The addition of supplementary nutrients for the acetic acid producing fermentation of Corni fructus enhances the rate of fermentation. In the process of Corni fructus vinegar production, the appropriate amount of Corni fructus extract to be added was found to be 6%, while the use of reducing sugars and total sugar including free sugar were very low.
Acetic acid production rate was lowered as the amount of Corni fructus extract added were increased. However, the production of acetic acid was significantly increased by the addition of supplementary nutrients. The addition of supplementary nutrients also lowered the residual alcohol content in the final product of vinegar. When the Corni fructus extract was used at a level of 2∼10%, total acidity of the product was no less than 5% and the acetic acid conten of 3.97∼5.25%. When the level of Corni fructus estract was increased to 8% and 10%, the acetic acid contents were decreased to 4.16% and 3.97%, respectively.
The color values of L, a and b were not changed significantly during fermentation, while the values were changed with the amount of Corni fructus extract used. Solid matter content of the Corni fructus vinegar was higher than that of commercial fruit vinegars, while lower than persimmon vinegar.
To obtain basic information for development of functional beverage and tea product using Corni fructus, physicochemical and functional characteristics of Corni fructus extracts were investigated. When Corni fructus was extracted with water at 80℃, turbidity and sweetness were highest with 8 or 10 hours of extraction. Yellowness and redness values of the extract were higher as the extraction time longer, but the lightness was not. Sensory evaluation of the extract showed that color, sourness, sweetness, astringency and flavor were generally highest with 8 or 10 hours of extraction. However, the overall acceptancy value was highest with 2 hours of extraction. According to above results, 8 hours of extraction was suggested to be adequate for the extraction.
Reduction of sourness and astringency of Corni fructus were investigated with addition of sugars, β-cyclodextrin, sweetmate and various spices. When sucrose and fructose were added to 1% solution of Corni fructus extract at a concentration of 0-20%, the sensory scores of sourness, astringency and sweetness were improved as the concentration of the sugars increased, obtaining highest score of overall acceptability at a concentration of 15%.
When β-cyclodextrin is added, the sourness and astringency were improved as the concentration of the β-cyclodextrin increased. However, the other sensory parameters were inferior to those added with sucrose or fructose. The sourness and astringency were improved also by sweetmate. but the other sensory parameters were inferior to those added with sucrose, fructose or β-cyclodextrin.
Masking flavors were also effective in reducing the sourness and astringency of the Corni fructus extract. Among the 5 commercial masking flavors tested in the present study, M.S 1998-0-69 and M.S SF-474A were the best in reducing the sourness and astringency.
To establish the direction and strategy to develop Corni fructus drink product, commercial drink products of domestic and Japan in popular were collected and characterozed. Domestic drink products were categorized into 4 types, products containing herb medicine, functional products, fruit juice products and carbonated drinks. Many of the drink products containing herb medicine had pH of 3.51∼4.0, sweetness of 10.01∼15.0 and taste of sweetness and bitterness.
The functional drink products had pH of 3.01∼3.5, sweetness of 10.01∼15.0 and taste of sourness and sweetness.
Among the potential competing products to the Corni fructus drink product to be developed in the present study, the most popular 4 products were found to be "Youg-Bi-Cheon", "Royal-D", "Un-Ji-Cheon", and "Maek-Saeng" in order. The age groups of the major consumers of the most popular 4 drink products were age groups of 40's and 30's.
From the informations on quality characteristics of commercial drink products and other informations, the direction of designing Corni fructus drink product was determined to be low in pH and sweetness to give freshness and to reduce bitter and sour tastes. The product is also to be added with sugars and vitamins to suit for the taste of the major target consumer groups of 30's and 40's as a health food.
For the formulation of Corni fructus drink product, sucrose and fructose was selected as sweeteners to reduce sour and astringent taste of Corni fructus.
Several functional ingredients, such as trehalose, taurine and L-carnitine were also used to suit for the public taste. Manufacturing process was established to be suitable for mass production and production on a commercial scale.
Tea and soft capsule type products of Corni fructus high in added value were developed under consideration of conveniency. For the formulation of tea product anhydrous glucose was used as a sweetener because it was satisfactory in textural behavior, easiness of working and sensory properties. For the capsule type product of Corni fructus, soybean oil, palm oil, lecithin, gelatin, glycerin, Blue No 1, Yellow No 5 and Red No 40 were used. Manufacturing processes of tea and capsule type product were established to be suitable for mass production and production on a commercial scale.
The Corni fructus products of drink, tea and soft capsule type were tested for quality. The appearance of the product, flavor, sensory characteristics, E. coli, viable cell count, disruption and solubility are satisfactory for 4 months of storage at low temperature (5℃), room temperature and high temperature (37℃).
Hydrogen donating activities of the methanol extract of Corni fructus and chloroform, ethyl acetate, butanol and water subfractions of the methanol extract were studied by measuring the removal of DPPH free radical. The methanol extract showed a dose of 50% removal (RD50) of 450㎍, while BHT, a reference antioxidant, showed the RD50 of 27㎍. Among the subfractions ethyl acetate fraction had the highest activity of DPPH radical removal showing RD50 of 85㎍, followed by butanol fraction of 310㎍, chloroform fraction of 490㎍. The concentrate of Corni fructus extract used for drink type and solid type food product also showed a dose dependent activity of DPPH radical removal.
Antioxidant activity of Corni fructus extract against linoleic acid autoxidation was observed in water extract and 70%, 90% and 100% ethanol extract. Among the subfractions of ethanol extract, ethyl acetate soluble fraction had the highest activity of antioxidant against linoleic acid. Ursolic acid isolated from the chloroform soluble fraction of Corni fructus ethanol extract also inhibited the autoxidation of linoleic acid at a concentration of 100㎍/㎖.
In an in vitro model system of lipid peroxidation using liver homogenate, in which lipid peroxidation is induced by H2O2 and FeSO4, methanol extract of Corni fructus significantly inhibited the lipid peroxidation in a dose-dependent manner. Among the subfractions of methanol extract, ethyl acetate soluble fraction showed the most potent antioxidant activity in the liver homogenate.
Antidiabetic functions of Corni fructus were investigated in hyperglycemic rats treated with streptozotocin. The streptozotocin-treated rats showed symptoms of polydypsia, polyphasia and polyuria, typical symptoms of severe diabetes. The diabetic rats had very low weight gain despite the polyphasia. The symptoms of polydypsia, polyphasia, polyuria and low growth, as well as hyperglycemia were improved by oral treatment of the methanol extract of Corni fructus and subfractions of the methanol extract. Among the subfractions of methanol extract, chloroform soluble fraction was most effective for improving the diabetic symptoms.
The streptozotocin-induced diabetic rats had significantly higher values of plasma GOT and GPT than normal rats, which was lowered by oral treatment of the methanol extract of Corni fructus and subfractions of the methanol extract.
Among the subfractions, butanol soluble fraction was most effective in lowering the plasma levels of GOT and GPT.
The diabetic rats had higher levels of plasma total cholesterol and LDL-cholesterol and lower level of HDL-cholesterol, consequently higher value of atherogenic index than the normal rats. The diabetic rats had also higher level of plasma triglyceride than the normal rats. The hyperlipidemia in diabetic rats was improved some by feeding Corni fructus or by oral administration of the methanol extract and subfractions of the methanol extract. The anti-hyperlipidemic effect of Corni fructus was found to be highest in the ethyl acetate soluble fraction.
The diabetes induced by streptozotocin came with fatty liver characterized by elevated level of liver cholesterol and triglycerides. The high level of liver lipids were lowered some by feeding Corni fructus or by oral administration of the methanol extract and subfractions of the methanol extract. The lipotropic effects of Corni fructus was found to be highest in the ethyl acetate soluble fraction.
Hypolipidemic functions of Corni fructus were investigated in rats fed with an atherogenic diet containing butter and cholesterol. The rats fed atherogenic diet had higher levels of plasma total cholesterol and LDL-cholesterol and lower level of HDL-cholesterol, consequently higher value of atherogenic index than the normal rats. The rats fed the atherogenic diet had also higher level of plasma triglyceride and liver lipids than the rats fed corn oil diet. The hyperlipidemia and fatty liver in rats fed atherogenic diet were improved some by feeding Corni fructus or by oral administration of the methanol extract and subfractions of the methanol extract. The anti-hyperlipidemic and lipotropic effects of Corni fructus were found to be highest in the ethyl acetate soluble fraction of methanol extract.
Extracts of distilled water, 70%, 90% and 100% ethanol were tested for the enhancement of immune function. When the extracts were added to culture medium of mouse spleen cell at concentrations of 1 ㎍/ml, 10 ㎍/ml and 100 ㎍/ml, only the 70% ethanol extract was observed to enhance proliferation of mouse spleen cell. When ConA or LPS was added to spleen cell culture medium along with the extracts, water extract inhibited the growth of cell, while ethanol extract stimulated the growth of spleen cell.
When the Corni fructus extracts of distilled water, 70%, 90% and 100% ethanol were tested for the growth of cancer cell at concentrations of 10 ㎍/ml, 100 ㎍/ml and 1000 ㎍/ml, the extracts suppressed growth of A549, a human lung cancer cell line, and MCF-7, a human breast cancer cell line, at a concentration of 1000 ㎍/ml.
To isolate active compounds of anti-cancer function, the methanol extract of Corni fructus was fractionated sequentially with hexane, chloroform, ethyl acetate, n-butanol and water. The hexane soluble fraction and chloroform soluble fraction showed cytotoxic activities against A549, a human lung cancer cell line, and MCF-7, a human breast cancer cell line in a dose dependent manner, while the other fractions didn't. The hexane fraction and chloroform fraction were subjected to silica gel column chromatography repeatedly. From the column chromatography of the fractions, two active compounds, compound 1 and 2, were isolated. Of the two active compounds, the compound 2 was identified to be ursolic acid according to the structural information of IR, mass spectroscopy and NMR.
Cytotoxic effects of the compounds isolated from hexane and chloroform fractions on cancer cells were investigated in vitro. The compound 1, a substance isolated from the hexane fraction, at a concentration of 10 ㎍/㎖, showed cytotoxicity of 41.66% against A549, 17.74% against MCF-7, and 33.71% against HL-60. The compound 2, isolated from the chloroform fraction showed 9.25%, 8.92% and 23.61% cytotoxicity against the human cancer cell lines of A549, MCF-7 and HL-60, respectively, at a concentration of 10 ㎍/㎖. Marked morphological changes in cancer cells were observed with the substances.
Nuclei were shrunk and the cells were observed to be detached from the plate when the cancer cell lines of A549 and MCF-7 were treated with the isolated compounds.
2. Suggestions for Application of the Results
The investigation of basic and processing technology for manufacturing functional food products of Corni fructus has been successfully carried out. The processing technology developed in the present study may be applied directly to other fruit type herbal medicine, such as Schisandrae fructus and Lycii fructus.
The acquired basic information and processing skills are suggested to be transferred to any demanding companies which have processing lines of wine, drink and tea, or to be industrialized through establishing processing plants in the local area.
목차 Contents
- 표지 ... 1
- 제출문 ... 2
- 요약문 ... 4
- SUMMARY ... 14
- CONTENTS ... 26
- 목차 ... 28
- 제1장 연구개발과제의 개요 ... 30
- 제2장 국내외 기술개발 현황 ... 32
- 제3장 연구개발수행 내용 및 결과 ... 34
- 제1절 산수유를 이용한 약용 발효주 개발 ... 34
- 1. 재료 및 방법 ... 34
- 2. 결과 및 고찰 ... 42
- 제2절 산수유를 이용한 발효음료 개발 ... 60
- 1. 재료 및 방법 ... 60
- 2. 결과 및 고찰 ... 63
- 제3절 산수유를 이용한 음료 개발 ... 74
- 1. 재료 및 방법 ... 74
- 2. 결과 및 고찰 ... 76
- 제4절 산수유를 이용한 차와 캡슐제품의 개발 ... 96
- 1. 재료 및 방법 ... 96
- 2. 내용 및 결과 ... 97
- 제5절 항 산화 활성 ... 104
- 1. 재료 및 방법 ... 104
- 2. 결과 및 고찰 ... 106
- 제6절 항 당뇨 활성 ... 112
- 1. 재료 및 방법 ... 112
- 2. 결과 및 고찰 ... 114
- 제7절 고지혈증 개선 효과 ... 124
- 1. 재료 및 방법 ... 124
- 2. 결과 및 고찰 ... 125
- 제8절 산수유 추출물 및 제품의 면역 활성 ... 134
- 1. 재료 및 방법 ... 134
- 2. 결과 및 고찰 ... 135
- 제9절 산수유 추출물 및 제품의 항암 활성 ... 142
- 1. 재료 및 방법 ... 142
- 2. 결과 및 고찰 ... 144
- 제4장 목표달성도 및 관련분야에의 기여도 ... 174
- 제5장 연구개발결과의 활용계획 ... 176
- 제6장 연구개발과정에서 수집한 해외과학기술정보 ... 178
- 제7장 참고문헌 ... 180
- 끝페이지 ... 184
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