초록

○ 연구결과
1. 생리활성물질 증진 및 안정된 수량확보가 가능한 재배기술 개발
2. Isothiocyanate가 2배이상 증진된 열무에서 항암효과가 인정되었음
3. 기능성열무로 제조한 열무김치의 경우 인체 위암세포와 결장암의 성장 저해효과 확인
4. 기능성열무를 소재로한 김치제조기술의 표준화 확립

Abstract ▼

Ⅱ. Results and discussion
Yeol Mu(Young radishes) were cultivated in the yellow soil with 1,818 g/m3 sulfur, 1,818 g/m³ sulfur + 2 cm soil covering, and in normal soil for control.
There were differences of leaf number and plant height by treatments. Yeol Mu cultivated in the yellow

Ⅱ. Results and discussion
Yeol Mu(Young radishes) were cultivated in the yellow soil with 1,818 g/m3 sulfur, 1,818 g/m³ sulfur + 2 cm soil covering, and in normal soil for control.
There were differences of leaf number and plant height by treatments. Yeol Mu cultivated in the yellow soil with sulfur showed lower growth and development, compared with control, the yield of that were 75% against control. But It seemed to be improved with proper temperature and water control. Upon investigation by interview, most of the young radish have been cultivated from the greenhouse which was less than 1ha and shipped to Department store or large discount store.
Judging from the acute oral toxicity test, there were no treatment-related effects on mortality, gross finding and so forth in ICR mice treated with a single oral dose of kimchi made from Yeol Mu cultivated with sulfur.
When Yeol Mu was sowed seeding in the yellow soil with sulfur in spring, summer and autumn, germination percentage was similar in each soil treatment, but showed lower level in autumn. This reason would be that temperature which gives the biggest effect in germination was lowest in autumn.
In demonstration at farmers field cultivation, Yeol Mu seed was purchased in the market. The seed size and the shape of the young radish were different in seed company. so the required seed quantity was different.
Germination percentage at 10th day after sowing was 87.6∼00% in cultivar, and leaf number or plant height was different in growth, also. the yield of Yeol Mu was highest in cultivar C that is 2,669㎏/10a , but statistical significantly different was not found in each variety.
The content of each sample's water, protein, lipid and ash were 89.5∼5.2,1.1∼.7, 0∼.4 and 1.1∼.9% respectively, and Mineral contents were different in the dried sample.
Young radishes which were grown in the soil containing sulfur increased quinine reductase (QR) activity in Hepa 1c1c cells and isothiocyanate-like compound analyzed by HPLC. QR inducing activity was maximum in young radishes grown with 1,818 g/㎥ulfur and was decreased when the soil was neutralized with lime mortar in order to improvea recovery. These results have suggested that consumption of young radishes, especially grown in the presence of sulfur, would prevent from cancer incidence through inducing detoxification enzymes and could have therapeutic effects for chemoprevention. The oral administration of extracts of young radishes cultivated with sulfur after intravenous tumor cell injection achieved a marked reduction of pulmonary colonization in mice. Treatment of the mice with extracts of young radish cultivated with sulfur did not show any increase in the number of CD8+ or NK T cells in the spleen, indicating no influence on host immunity. Sulforaphane, which could be a candidate for an active compound from young radishes cultivated with sulfur, inhibited cell growth of B16-F10 melanoma cells. In addition, extracts of the young radish cultivated with sulfur-fed group showed enhanced quinine reductase (QR) activities in the liver and lung and a slight increase of glutathione S-transferase (GST) activity in the liver. These results have suggest that the administration of extracts of young radishes cultivated with sulfur suppressed pulmonary tumorigenesis, possibly due to increased the activity of detoxification enzymes in the liver and lung and partly due to cell cytotoxicity. The oral administration of extracts of young radishes cultivated with sulfur after subcutanepous CT26 tumor cell injection achieved a marked reduction of tumor size in mice. However, no effect were observed with extracts of fermented pickle using young radishes cultivated with sulfur in both murine models.
Manufacturing method and ingredient ratios for young radish(YR) kimchi and young radish mul(YRW) kimchi were standardized from literatures. Fermentation properties and in vitro anticancer effects of YR kimchi and YRW kimchi were investigated during fermentation at 5℃. Ingredients having frequency of use greater than 50% were only used in the standardization process. Green onion, red pepper, red pepper power, garlic, ginger, and anchovy juice were included in YR kimchi. Green pepper, red pepper, garlic, ginger, and starch were included in YRW kimchi. Standardized ingredients ratio of YR kimchi on young radish 100 g was as follows: green onion 8.0±ｱ3.8 g, crushed garlic 2.9±ﾜ1.3 g, crushed ginger 1.6±0.7 g, red pepper 7.0±21.7 g, red pepper powder 4.2±`1.2 g, and anchovy juice 3.7±0.5 g.
Standardized ingredients ratio of YRW kimchi on added water 100 was as follows: young radish 50.6±10.8 g, crushed garlic 3.0±<0.7 g, crushed ginger 1.5±g0 g, green onion 3.3±1.3 g, green pepper 3.3±ｺ1.9 g, red pepper 2.4±1.3 g, and starch 1.5±0.6 g. Fermentation of YR kimchi during 2-3 weeks led to the decrease of pH down to pH 4.3, the increased acidity, and the highest Leuconostoc sp. counts. YR kimchi showed the acidity of 1.04-1.27% at the pH 4.3, when the kimchi was ripened properly. Fermentation of YRW kimchi during 9 days led to the decrease of pH down to pH 4.3 and the acidity of 0.20%. Inhibitory effects of YR kimchi and YRW kimchi juices on the growth of AGS human gastric adenocarcinoma cells in MTT assay were increased with the added concentration. On considering of moisture contents, YR kimchi might be showed similar inhibitory effects on the growth of AGS human gastric and HT-29 human colon adenocarcinoma cells in MTT assay to YRW kimchi, which the inhibition rates were greater than 50%.
YR kimchi juice showed a higher inhibitory effect on the growth of AGS human gastric adenocarcinoma cells than that of Bae Chu(Korean chinese cabbage) kimchi at same concentration. That is, YR kimchi and Bae Chu kimchi juices indicated inhibition rates of 50% and 58%, respectively.
Young radishes (YR, Yeol Mu) were cultivated in the soil with and without sulfur. YR-control, YR-A and YR-B samples were grown in the soil without sulfur, with 1,818 g/m3 sulfur and 1,818 g/m3 sulfur + soil 2 cm, respectively. YR kimchies were prepared using YR-control, YR-A and YR-B. Fermentation properties of the YR-kimchis were investigated during fermentation at 5℃ for 8 weeks. In vitro growth inhibitory effects of YR samples and YR-kimchis were also investigated in HT-29 human colon and AGS human gastric adenocarcinoma cells. In vivo antitumor effects of methanol extracts from the young radish kimchies were investigated in sarcoma-180 cell transplanted mice. Juices from YR-A and YR-B showed higher growth inhibitory effects in AGS human gastric and HT-29 human colon adenocarcinoma cells than the juice from YR-control at the same concentration. The inhibitory effect of YR-A was similar to that of the YR-B. YR kimchis after 4-5 weeks at 5℃ showed higher acidity of 0.88-1.20% with pH 4.3-4.5 and the kimchis kept approximately pH 4.0 until 8 weeks. The kimchi-A and kimchi-B showed faster fermentation time, higher levels of Leuconostoc sp. and lower of Lactobacillus sp. during the fermentation, comparing to the control kimchi. The YR-A and YR-B juices exhibited higher inhibitory effects(88% and 87%) on the growth of HT-29 human colon adenocarcinoma cells than that of the control kimchi(59%) at the higher concentration of 20 ㎕assay.
The YR-A and YR-B juices also exhibited higher inhibitory effects(74%) on the growth of AGS human gastric adenocarcinoma cells than that of the control kimchi(57%) at the higher concentration of 20 ㎕assay. Methanol extracts from the kimchies also led to the similar results to the those of the juices. Young radish kimchis inhibited the growth of cells in a concentration and time dependent manner. YR-A induced apoptosis as determined by DAPI staining and showed the increased Bax and decreased Bcl-2 in expression of active anticancer compounds, comparing to control kimchi. These results suggested that preparing of kimchi using differently cultivated YR especially in the presence of sulfur, which can help to synthesize sulfur-containing compounds, could increase the anticancer effect in vitro. Methanol extracts from the young radish kimchies were investigated in sarcoma-180 cell transplanted mice. The solid tumor growth was inhibited most effectively by the kimchi prepared using young radish grown in the soil with sulfur, especially YR-B. The treatment of the methanol extracts from YR-A and YR-B induced the high glutathione S-transferase (GST) activities and glutathione contents, comparing to control kimchi. These results suggested that the antitumor effects of young radish kimchi might be enhanced by using young radish cultivated in the presence of sulfur, which can help to produce sulfur-containing compounds in young radish. In order to investigate effect of the Yeol Mu cultivated in the sulfur containing soil on a texture and taste of kimchi, Yeol Mu kimchi with different ingredients and grades were prepared. The effects were determined from the results of sensory evaluation test. From the results of sensory evaluation test, increased ingredients content induced the good acceptability as kimchi and anticancer effects. Therefore, since kimchi added total ingredients in ratios of 1:1.5 improved the functional properties and tastes of kimchi, it is possible to develop manufacturing method of Yeol Mu kimchi with improved functionality and good taste.

목차 Contents

• 표지 ... 1
• 제출문 ... 3
• 요약문 ... 4
• SUMMARY ... 7
• CONTENTS ... 11
• 목차 ... 16
• 제 1 장 연구개발과제의 개요 ... 21
• 제1절 연구개발 목표와 내용 ... 21
• 제 2 장 국내외 기술개발 현황 ... 23
• 제 3 장 연구개발수행 내용 및 결과 ... 24
• 제1절 기능성 열무 생산기술 연구 ... 24
• 1. 연구수행 방법 ... 24
• 가. 유황처리 함량 및 방법에 따른 생육반응 연구 ... 24
• 나. 재배시기에 따른 생산성 검증 ... 26
• 다. 생산현장 실증시험 ... 26
• 2. 연구결과 ... 27
• 가. 유황처리 함량 및 방법에 따른 생육반응 연구 ... 27
• 1) 발아세 ... 27
• 2) 상토 이화학성 ... 27
• 3) 식물체 무기성분 ... 28
• 4) 처리별 초장 및 엽수의 경시적 변화 ... 28
• 5) 생육 및 수량 ... 29
• 6) 열무재배 및 생산 실태조사 ... 30
• 7) 마우스를 이용한 급성경구독성시험 ... 32
• 나. 재배시기에 따른 생산성 검증 ... 34
• 1) 발아율 ... 34
• 2) 상토 이화학성 ... 34
• 3) 식물체 무기성분 ... 35
• 4) 재배시기 및 토양처리별 엽수 ... 35
• 5) 생육 및 수량 ... 37
• 다. 생산현장 실증시험 ... 38
• 1) 품종별 종자 소요량 ... 38
• 2) 재배지 토양특성 ... 38
• 3) 열무 품종에 따른 생육 ... 38
• 4) 수확시 품종별 생육 및 수량 ... 39
• 5) 농가 생산시험 ... 40
• 6) 열무 품종별 성분분석 ... 41
• 제2절 유용물질 탐색 및 추출 ... 43
• 1. 연구수행 방법 ... 43
• 가. 유황처리 농도별 열무의 이화학적 특성 및 유용물질탐색 ... 43
• 1) 열무의 유효성분의 추출 ... 43
• 2) quinone reductase (QR)의 inducer potency ... 43
• 3) Isothiocyanate와 glucosinolate의 측정 ... 44
• 4) melanoma tumor development ... 44
• 2. 연구결과 ... 44
• 가. 유황처리 농도별 열무의 이화학적 특성 및 유용물질탐색 ... 44
• 1) 생육조건에 따른 열무에서 quinone reductase inducing activity의 측정 ... 44
• 2) 생육시기와 재배조건에 따른 열무에서 quinone reductase inducing activity의 측정 ... 47
• 3) 열무품종별과 재배조건에 따른 열무에서 quinone reductase inducing activity의 측정 ... 49
• 4) 유황처리 열무에서 유용물질의 탐색 ... 49
• 5) 유황처리열무의 유용물질의 물리, 화학적 특성조사 ... 51
• 6) mouse melanoma model에서 열무추출액의 효과 ... 52
• 7) 열무추출액중의 active compound인 isothiocyanate의 효과 : B16F10 melanoma cell에 대 한 cytotoxicity ... 54
• 8) 쥐에 장기 투여시의 열무추출액의 효과 ... 55
• 9) CCl4-induced liver toxicity model에서 열무추출액의 효과 ... 56
• 10) 유황열무추출액중의 active compound인 isothiocyanate의 효과 : 파골세포분화 ... 57
• 11) 열무김치의 in vivo 활성 (murine melanoma model) ... 62
• 12) 열무김치의 in vivo 활성 (murine colon cancer model) ... 65
• 13) 유황열무추출액의 in vivo 활성 (murine colon cancer model) ... 68
• 14) 유황열무 in vivo 투여시 간에서 phase II enzyme의 유도 ... 69
• 제3절 기능성 열무를 이용한 김치제조 기술개발 및 상품화 연구 ... 71
• 제 1 세부 열무김치 제조방법의 표준화 ... 71
• 1. 연구수행방법 ... 71
• 가. 열무의 영양성분 분석 및 열무김치 제조의 표준화 ... 71
• 나. 표준화 열무김치의 생리기능성 검증 ... 72
• 2. 연구결과 ... 75
• 가. 열무김치의 표준화 연구 ... 75
• 1) 문헌에 의한 열무김치 및 열무물김치의 표준화 ... 75
• 2) pH 및 산도의 변화 ... 77
• 3) 젖산균 수의 변화 ... 79
• 나. 토양의 유황처리 농도 및 방법별 열무김치 기능성 연구 ... 81
• 1) 기능성 열무의 일반 성분 분석 ... 81
• 2) 일반 열무, 열무김치, 열무물김치의 암세포의 성장 억제 효과 ... 82
• 3) 발효과정 중 기능성 열무물김치 pH 및 산도의 변화 ... 83
• 4) 발효과정 중 기능성 열무물김치의 젖산균 수의 변화 ... 84
• 5) 기능성 열무 및 열무물김치의 항돌연변이 효과 ... 86
• 6) 기능성 열무의 항암 효과 ... 87
• 7) 기능성 열무물김치의 항암 효과 ... 88
• 제 2 세부 가공방법 및 가공제품개발 ... 91
• 1. 연구수행방법 ... 91
• 가. 기능성 열무를 이용한 열무김치 및 열무물김치의 제조 표준화 ... 91
• 나. 표준화 열무김치의 생리기능성 검증 ... 92
• 다. 재배시기별로 수확한 열무를 이용하여 제조한 열무김치 및 열무물김치의 in vitro의 항암효과검토 ... 93
• 라. 기능성열무로 제조한 열무김치의 세포독성 및 in vivo 항암효과검토 ... 94
• 2. 연구결과 ... 96
• 가. 열무김치의 표준화 연구 ... 96
• 1) 열무김치의 재료 배합비 및 제조 표준화 ... 96
• 나. 여름에 파종한 열무로 제조한 기능성 열무김치 및 열무물김치의 발효 특성 ... 99
• 1) pH 및 산도의 변화 ... 99
• 2) 젖산균 수의 변화 ... 102
• 다. 가을에 파종한 열무로 제조한 기능성 열무김치 및 열무물김치의 발효 특성 ... 105
• 1) pH 및 산도의 변화 ... 105
• 2) 젖산균 수의 변화 ... 108
• 라. 기능성 열무로 제조한 열무김치 및 열무물김치의 세포 독성 및 in vitro 항암 기능성 연구 ... 112
• 1) 열무김치 및 열무물김치의 세포 독성 효과 및 항돌연변이 효과 ... 112
• 2) 여름에 파종하여 재배한 열무로 제조한 열무김치 및 열무물김치의 항암효과 ... 113
• 3) 가을에 파종하여 재배한 열무로 제조한 열무김치 및 열무물김치의 항암효과 ... 116
• 마. 기능성 열무로 제조한 열무김치의 세포 독성 및 in vivo 항암 기능성 연구 ... 118
• 1) 열무김치 및 열무물김치의 세포독성 및 고형암 성장 저지 효과 ... 118
• 2) 열무김치 및 열무물김치의 마우스의 장기무게 변화 ... 118
• 3) 열무김치 및 열무물김치의 Glutathione S-transferase (GST) 활성 ... 119
• 제 3 세부 가공 공정 확립 및 산업화 제품 개발 ... 120
• 1. 연구수행방법 ... 120
• 가. 기능성 열무를 이용한 열무김치의 제조 ... 120
• 나. 기능성 열무로 재배한 열무김치의 색소 및 비타민 C 함량 ... 121
• 다. 기능성 열무로 재배한 열무김치의 기능성 증진효과 ... 121
• 라. 품종별로 수확한 열무를 이용하여 제조한 열무김치 in vitro 항암효과검토(MTT test, cell growth inhibition test) ... 123
• 2. 연구결과 ... 124
• 가. 부재료 배합에 따른 유황처리 열무김치의 개발 ... 124
• 1) 유황처리 열무김치의 제조 연구 ... 124
• 2) 마늘, 고춧가루, 멸치액젓의 조절에 따른 이화학적 특성 ... 125
• 3) 마늘, 고춧가루, 멸치액젓의 조절에 따른 관능검사 ... 127
• 4) 부재료 첨가 농도에 따른 유황처리 열무김치의 암예방 효과 ... 129
• 나. 기능성 열무로 재배한 열무김치의 색소 및 비타민 C 함량 ... 133
• 1) Carotenoids 함량 ... 133
• 2) Chlorophylls 함량 ... 133
• 3) Vitamin C 함량 ... 133
• 다. 유황처리 열무의 품종에 따른 열무김치제조 연구 ... 134
• 1) 품종에 따른 유황처리 열무의 김치제조 연구 ... 134
• 2) 유황처리 열무 품종에 따른 이화학적 특성 ... 134
• 3) 유황처리 열무 품종에 따른 관능검사 및 일반열무김치와의 비교 ... 136
• 4) 재료 배합비에 따른 열무김치의 암예방 효과 ... 140
• 5) 유황처리 열무김치의 품종에 따른 암예방 효과 비교 ... 144
• 라. 유황처리 농도를 달리하여 재배한 열무로 만든 김치의 in vitro 항암효과 및 apoptosis ... 145
• 1) 세포독성 작용 및 고형암 성장 억제 효과 ... 145
• 2) 마우스의 장기무게 변화 ... 147
• 3) 간에서 GST활성 및 GSH 함량의 변화 ... 147
• 제 4 장 목표달성도 및 관련분야에의 기여도 ... 153
• 제1절 기능성 열무 생산기술 연구 ... 153
• 제2절 유용물질 탐색 및 추출 ... 153
• 제 3 절 기능성 열무를 이용한 김치제조 기술개발 및 상품화 연구 ... 154
• 제 5 장 연구개발결과의 활용계획 ... 155
• 제 6 장 연구개발과정에서 수집한 해외과학기술정보 ... 156
• 제 7 장 참고문헌 ... 157
• 끝페이지 ... 160