보고서 정보
주관연구기관 |
목포대학교 Mokpo University |
연구책임자 |
김관수
|
참여연구자 |
박시형
|
보고서유형 | 최종보고서 |
발행국가 | 대한민국 |
언어 |
한국어
|
발행년월 | 2007-02 |
과제시작연도 |
2005 |
주관부처 |
농촌진흥청 |
사업 관리 기관 |
농촌진흥청 Rural Development Administration |
등록번호 |
TRKO201100015737 |
과제고유번호 |
1390004478 |
사업명 |
농업생명공학기술개발 |
DB 구축일자 |
2013-04-18
|
초록
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○식량작물의 주요 영양 및 생리활성물질의 정량적 표준화 분석기법 개발
○식량작물의 주요 영양 및 생리활성물질 대량분석기법 개발 및 실용화
○식량작물 주요품종 및 유전자원의 영양 및 생리활성물질 특성평가
○국가보존 특용 및 약용유전자원의 기능성 성분 중심으로 품질특성 평가
○표준화된 성분분석방법과 대표적인 기능성성분을 제시 및 정량자료 작성
○성분특성이 우수한 유전자원을 선발하여 품종개량 및 기능성 제품 개발소재로서 유전자원의 활용도 증대
○약용 및 특용작물의 품질 특성을 신속히 평가할 수 있는 화학성
○식량작물의 주요 영양 및 생리활성물질의 정량적 표준화 분석기법 개발
○식량작물의 주요 영양 및 생리활성물질 대량분석기법 개발 및 실용화
○식량작물 주요품종 및 유전자원의 영양 및 생리활성물질 특성평가
○국가보존 특용 및 약용유전자원의 기능성 성분 중심으로 품질특성 평가
○표준화된 성분분석방법과 대표적인 기능성성분을 제시 및 정량자료 작성
○성분특성이 우수한 유전자원을 선발하여 품종개량 및 기능성 제품 개발소재로서 유전자원의 활용도 증대
○약용 및 특용작물의 품질 특성을 신속히 평가할 수 있는 화학성분 분석기법 및 기능성 평가 기법 개발
○화학성분 및 기능성 평가기법을 토대로 우수품종 육성 지표로 사용하여 수집 유전자원의 평가 및 선발
○농업유전자원으로부터 신 기능성 물질 분리 및 이용기술 개발 (항산화물질, 항당뇨활성물질)
○쌀 유전자원의 이화학적 특성 및 생리활성 평가
○쌀 유전자원의 가능성 식품 개발
○평가를 통해 선발된 기능성 식용작물 및 특약용작물의 제품적 특성 연구
○생리활성 및 제품적 특성이 고려된 유전자원의 활용방안 탐색 및 성분평가
Abstract
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1. Quality evaluation of food crops (soybean)
Because of its unique chemical composition, Soybean is a very valuable and useful agricultural commodity. Among legumes and cereals, it has the highest protein content and the second highest oil content. Other valuable components that are important t
1. Quality evaluation of food crops (soybean)
Because of its unique chemical composition, Soybean is a very valuable and useful agricultural commodity. Among legumes and cereals, it has the highest protein content and the second highest oil content. Other valuable components that are important to human and animal health include fatty acids, oligosaccharides, saponin, and isoflavones. However, mass analytical method of these constituents are not well established to quantify. Therefore, the purpose of this study was to established the appropriate method of extraction, isolation, sample preparation and determination of fatty acids, isoflavones, and oligosaccharides, and to developed the mass analytical method of these constituents using NIRS in soybean. Also, the 1,352 soybean germplasms were investigated their functional compounds, such as isoflavones, oligosaccharide and fatty acids. Likely many other oils of plant origin, most fatty acids in soybean are unsaturated. The highest percentage of fatty acid in soybean was linoleic acid(18:2), followed in a decreasing order by oleic(C18:1), palmitic(C16:0), linolenic(C18:3), and stearic acid(C18:0). The composition of C18:1 and C18:2 fatty acids among 517 soybean germplasms which were positively correlated with nutritional quality of edible vegetable oil, were ranged 13.29 ~ 43.30% and 38.21~61.51%, respectively, and the C18:3 which were negatively correlated with flavor quality were varied from 5.03% to 11.48%. Also, the composition of C16:0 and C18:0 which were known to raise total cholesterol levels in human serum were ranged 8.5 0~14.78% and 1.86~4.78% respectively. Among the 563 germplasms tested, there are significant correlations among unsaturated fatty acids: negative between C18:1 and C18:2(r=-0.961$^{**}$) or C18:3 (r=-0.756$^{**}$), and positive between C18:2 and C18:3(r=0.608$^{**}$). Also, the C18:1 was negatively correlated with saturated fatty acids, C16:0(r=-0.231$^{**}$) and C18:0(r=-0.479$^{**}$). The twelve isoflavones were separated from soybean extracts using LC/ES-MS. The chemical structures were elucidated as daidzein, glycitein, genistein, daidzin, glycitin, genistin, malonyldaidzin, malonylglycitin, malonylgenistin, acetyldaidzin, acetylglycitin, and acetylgenistin. The daidzein, glycitein, genistein and total isoflavone contents of 273 soybean germplasms collected in Korea, were ranged 26~1,459㎍/g, 0~235㎍/g, 129~1,379㎍/g, and 289~2917㎍/g, respectively. The effective acid hydrolysis conditions of isoflavone was investigated. Total isoflavone content was increased up to 3 hours at 1.0N HCI concentration and 100℃ oven heating. Therefore, the optimal hydrolysis conditions of soybean isoflavone was 3 hours hydrolysis using 1.0N HCI in 100℃ dry oven. To identify the soluble carbohydrates in soybean seeds, On-line HPLC-RID-ES/MS was used. The monosaccharide (glucose) and three oligosaccharides(sucrose, raffinose, and stachyose) were identified in Korean soybeans by their chromatographic behavior and results of on-line HPLC-RID-MS with Electrospray Ionization mode. In 485 soybean germplasms, the ranges of glucose, sucrose, raffinose, and stachyose were 0.03~0.98%, 2.33~6.96%, 0.08~1.87% and 0.75~3.18%, respectively. Among soybean samples, the composition ratio of soluble carbohydrates in soybean was high in the order of sucrose (58.01± 5.82%) > stachyose (29.80± 4.54%) > raffinose (10.13± 2.28%) > glucose (2.07± 1.75%). Sucrose appeared to be most prevalent in soybean soluble carbohydrates. The applicability of near infrared reflectance spectroscopy (NIRS) was tested to determine the fatty acid composition, isoflavone, and oligosaccharide contents in ground soybean seeds. The NIRS equation seems reliable for determining the isoflavones(total isoflavones, genistein, daidzein), fatty acidsColeic, linoleic, and linolenic acid), and oligosaccharides(total oligosaccharide, and sucrose) and NIRS method could be used as a mass screening method of soybean seed constituents.
2. Quality evaluation of oil crops and medicinal crops
This study was peformed to evaluate quality of germplasm including sesame. perilla, rapeseed, and medicinal crops. Lignan compounds were separated and used standard compound for quantitative analysis of sesame germplasm. In oil crops, oil content and fatty acids in germplasm from GeneBank, Korea were analyzed using GC and gravimetric method. In addition, nondestructive NIR methods were developed and validated for lignan, oil, and fatty acids. The results indicated that NIRS could be used to rapidly determine lignan and oil content, and fatty acid composition in sesame, perilla and rapeseed seeds in the breeding programs for development of high quality seeds. In addition, breeding lines of Atractylodes species and collected lines of Curcuma longa L were chemically evaluated. Chemical information obtained in this study were provided for D/B data to GeneBank and Rural Development Administration. Sesame (Sesamum indicum L); Sesame seed contains a number of antioxidants such as lignan and tocopheroL Among lignan compounds in sesame seeds, water-soluble lignan glucosides included sesaminol glucosides, sesamolinol glucosides, and pinoresinolglucosides. In this study, sesaminol triglucoside (STG), sesamolinol diglucoside, sesaminol (1→2) diglucoside and its isomer as (1-6) diglucoside were isolated using open chromatography and preparative high performance liquid chromatography CHPLC), and identified their structures by spectroscopic methods such as liquid chromatography and mass spectroscopy (LC-MS) and nuclear magneticresonance (NMR). Using these purified lignan glucosides as standard compounds, new HPLC analytical method to determine simultaneously four compounds was developed and ninety-three sesame germplasms were evaluated, resulting that average contents showed 68.8 ± 32.3, 3.6 ± 2.1, 19.9 ± 12.2, and 14.9 ± 7.9 mg/100 g seeds of STG, sesaminol (1→2) diglucoside, sesamolinol diglucoside, sesaminol (1→6) diglucoside, respectively. Near infrared reflectance spectroscopy (NIRS) was used to develop a rapid and nondestructive method for the determination of lignan contents on intact sesame seeds. Ninety-threeintact seeds were scanned in the reflectance mode of a scanning monochromator. This scanning procedure did not require the pulverization of samples allowing each analysis to be completed within minutes. Reference values for lignan contents were obtained by high performance liquid chromatography (HPLC) analysis. Calibration equations for lignans (sesamin, sesamolin) and lignan glycosides (sesaminol triglucoside and sesaminol diglucoside) contents were developed using modified partial least square (MPLS) regression with internal cross validation (n = 63). The equations obtained had low standard error of cross-validation (SECV) and moderate R² (coefficient of determination in calibration). Prediction of external validation set (n = 30) showed significant correlation between reference values and NIRS predicted values based on the SEP (standard error of prediction), bias, and r² (coefficient of determination in prediction). The models developed in this study had relatively higher values (more than 2.0) of SD/SEP(C) for all lignans and lignan glycosides except for sesaminol diglucoside having a minor amount, indicating good correlation between reference and NIRS estimate. Perilla (Perilla frutescens var. japonica (Hassk.) Hara); A total of 397 samples (about 2 g of intact seeds) were scanned in the reflectance mode of a scanning monochromator, and the reference values for the oil content and fattyacid composition measured by gravimetricmethod and gas-liquid chromatography, respectively. Calibration equations for oil and individual fatty acids were developed using modified partial least square regression with internal cross validation (n = 297). The equations for oil, oleic, and linolenic acid had lower SECV (standard errors of cross-validation), higher R²(coefficient of determination in calibration), and higher 1-VR (ratio of unexplained variance divided by variance) values than those for palmitic, stearic, and linoleic acid. Prediction of an external validation set (n = 100) showed significant correlation between reference values and NIRS estimated values based on the SEP (standard error of prediction), r² (Coefficient of determination inprediction), and the ratio of standard deviation (SD) of reference data to SEP. The models for oil content and major fatty acids, oleic and linolenic acid had relatively higher values of SD/SEP(C) and r²(more than 3.0 and 0.9, respectively), thereby characterizing those equations as having good quantitative information, while those of palmitic, stearic, and linoleic acid had lower values (below 2.0 and 0.7, respectively), unsuitable for screening purposes. Rapeseed (Brassica napus L.); A total of 349 samples (about 2 g of intact seeds) were scanned in the reflectance mode of a scanning monochromator, and the reference values for fatty acid composition were measured by gas-liquid chromatography. Calibration equations for individual fatty acids were developed using the regression method of modified partial least squares with internal cross validation (n = 249). The equations had low SECV (standard errors of cross-validation), and high R² (coefficient of determination in calibration) values (more than 0.8) except for palmitic and eicosenoic acid. Prediction of an external validation set (n = 100) showed significant correlation between reference values and NIRS estimated values based on the SEP (standard error of prediction), r² (coefficient of determination in prediction), and the ratio of standard deviation (SD) of reference data to SEP. The models developed in this study had relatively higher values (more than 3.0 and 0.9, respectively) of SD/SEP(C) and r² for oleic, linoleic, and erucic acid, characterizing those equations as having good quantitative information. In addition, breeding lines (n=119) of Atractylodes species and collected lines of Curcuma longa L. were chemically evaluated for contents of atrcatylon and curcumin, respectively, in roots, using HPLC.
3. Isolation and use of new functional substance from agricultural germplasm
Three compounds were isolated from AtractyZodes japonica MeOH extract and their structures were determined as actratylon (1), atractylenolide II (2) and 3-hydroxyatractylenolide III 3-0-glucoside (3). LC-MS analysis method was established using isolated compounds as references. Polar and nonpolar compounds were detected by APCI method, but nonpolar compound such as an atractylon (1) was not detected by ESI method. Positive ion detection mode was more sensitive than negative ion mode. Isolated compounds were used as reference compounds for evaluation of genetic resources. Two a -glucosidase inhibiting compounds were isolated from Codonopsis lanceolata and their structures were determined as adenosine (4) and tangshenoside I (5). Isolation of Tangshenoside I was first report in Codonopsis lanceolata. IC50 of adenosine (4) and tangshenoside I (5) against a-glucosidase were 9.3 and 1.4 mM, respectively. Two compounds were isolated from Adenophora triphylla var. japonica and their structure were determined as methyl adenophorate (6) and coniferyl alcohol glucoside (7). Isolation of coniferyl alcohol glucoside was first report in Adenophora triphylla var. japonica. These two compounds were used as reference compounds for identification of this plant. Three polyhydroxylated alkaloids were isolated from aqueous fraction of Adenophora triphylla var. japonica. The structures of isolated compounds were determined as 1-deoxynojirimycin (DNJ, 8), 1-deoxymannojirimycin (DMJ, 9) and 2R,5R-bis(hydroxymethyl)-3R,4Rdihydroxypyrrolidine (DMDP, 10). These compounds showed strong a-glucosidase inhibitory activitiy and DMDP showed most potent activity. IC50 of DNJ, DMJ and DMDP were 230 μM, 210 μM and DMDP 2.9 μM, respectively. Five flavonoid glycosides were isolated from Vigna radiata MeOH extract. Structures of isolated compounds were determined as vitexin (11), isovitexin (12), pueararin (13), daidzin (14) and pueraria glycoside 3 (15). These flavonoid glycosides showed lower DPPH radical scavenging activity than quercetin. In LC-MS analysis, these flavonoid glycosides were well detected in ESI negative ion mode.
4. Development of functional foods using elite germplasm
The objectives of the fourth sub-project were three as followed; firstly, to screen bioactivities of functional genetic resources, particularly rice resources, secondly, to develop the diverse functional foods using selected predominant genetic resources, and finally, to show actual and positive schemes for consumption of the genetic resources which had superior merits on health and well-being trend. The contents of the fourth sub-project were mainly divided into three parts. The first one was to search for the functionality data of genetic resources and to screen and evaluate bioactivities In particular, we focused on the functionality of rice resources, and monitored the crude fiber content of 394 species and built up the database. The second one was to evaluate the processing compatibility and bioactivity of selected resources by screening, and we laid stress on two crops; rice(Goamy 2) and sesame in this study. The third one was to develop various functional foods based on above processing compatibility and functionality data and to examine changes of bioactivity of the processed products. In this study, we collected the physicochemical characteristics and functionality data of wide ranged rice resources, and especially, examined the crude fiber content of 394 rice species. Besides, we set up the database based on the crude fiber and amylose content of rice resources. Crude fiber and soluble/nonsoluble fiber level of Goamy 2 were 1.5 to 2 times higher than those of Ilpum. Furthermore, we evaluated the processing compatibilities such as water absorbing characteristics, glucose content from rice by starch degrading enzyme(glucoamylase), and thermal property(gelatinization and retrogradation). In the gelatinization respects, Goamy 2 had dramatically high cool viscosity and low setback values although low peak viscosity and hot viscosity of Goamy 2 were remarkably low. In addition, the cooking property and texture of Goamy 2 were of low grade compared with those of other rices. The development of functional foods prepared with Goamy 2 was attempted with Paesulki and the Korean white gruel. The optimum ratio of Paesulki preparation was 125 g of Goamy 2, 30 mL of water, and 60 g of sugar by response surface methodology. And also, we conducted to estimate on functionality of sesame with roasting time. Lignan content, 1,1-Diphenyl-2-Picrylhydrazyl(DPPH) radical scavenging activity, and AI(antioxidant index from rancimat method) of roasted sesame extract significantly increased as the roasting time of sesame increased. We made a try of developing the functional food with Goamy 2 and sesame, and we suggested two types; black sesame gruel and sesame cookie. Moreover, there were few changes of bioactivity of processed products after cooking. In conclusion, these results showed the positive aspects of application, development, and commercialization of selected genetic resources from this project. We investigated on the functionality and processing compatability, and developed four functional foods such as Paesulki, the Korean white gruel, black sesame gruel, and sesame cookie with Goamy 2 and sesame. In spite of high functionality, Goami 2 had a poor processing compatability and a low sensory acceptance. We, however, made up for this weak points through processing and masking of sensory off-flavor. Unfortunately, we could not try investigating on the functionality of other genetic resources because of the limited period of research. Therefore, we look forward to studying on the bioactivities of extensive genetic resources and to developing more diverse functional foods.
목차 Contents
- 표지...1
- 제출문...3
- 요약문...5
- SUMMARY...9
- CONTENTS...14
- 목차...15
- 제 1 장 서론...17
- 제 2 장 국내외 기술개발 현황...18
- 제 3 장 연구개발수행 내용 및 결과...23
- 제 1 절 식량작물 유전자원의 품질특성평가...23
- 제 2 절 특약용작물 유전자원의 품질특성 평가...50
- 제 3 절 농업유전자원으로부터 신기능성 물질의 분리 및 이용...75
- 제 4 절 기능성 유전자원을 이용한 제품적 특성 연구...97
- 제 4 장 연구개발 달성도 및 대외기여도...163
- 제 1 절 연구개발목표 달성도...163
- 제 2 절 대여기여도...165
- 제 5 장 연구개발결과의 활용계획...165
- 제 6 장 기타 중요 변동사항...165
- 제 7 장 참고문헌...166
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