초록

○ 콩 이용 미생물 발효에 의한 CLA 함유 콩 발효물 제조
- CLA 생성능이 우수한 버섯 균주 선발 및 고체 발효기술 확립
- 공액리놀레산 및 가바, 비배당체 이소플라본 동시 증진 기술 개발
○ CLA 함유 콩 발효물의 체지방 감소 효과 검정
- 12주간 고지방 식이동물 시험에서 29% 체중감소 효과 확인
- 혈중 콜레스테롤 24%, LDL-콜레스테롤 49% 개선

(출처 : 보고서 요약서 p.3)

Abstract ▼

Results
Study on effect of body fat reduction of CLA-containing fermented soybean and its commercialization Due to a rapid social change and growth, modern people have been exposed to excessive stress and unhealthy environments. Especially, with the increase in average income, people tend to eat

Results
Study on effect of body fat reduction of CLA-containing fermented soybean and its commercialization Due to a rapid social change and growth, modern people have been exposed to excessive stress and unhealthy environments. Especially, with the increase in average income, people tend to eat foods containing high in cholesterol and fat.

This tendency causes an increase in various kinds of adult diseases.

Conjugated linoleic acid (CLA) exhibits many biological effects such as anticarcinogenic activity, immune stimulation, body fat reduction, cholesterol in blood, and lowering atherosclerosis. CLA is widely found in various foods, including dairy products, and some seafoods, but its contents ranging from 0.01 to 1.7%. The objective of this study was to develop of functional food material using soybean (Glycine max L.) having diet effect. Linoleic acid was included over 10% in soybean. So, we used linoleic acid to convert conjugated linoleic acid.

A. Development of quality control technology of CLA-containing fermented soybean

During the fermentation of soybean to produce CLA, the isoflavone-glycosides were decreased slowly, which was accompanied by the accumulation of aglycones. In fermented soybean, the isoflavone aglycones contents increased about to approximately 2.4 fold of their initial amounts, but the isoflavone-glycoside contents decreased from over 80%. And CLA were produced 32.8 mg/100 g in the fermentation of uram by matsutake mycelium. The antioxidant activity of the fermented soybean included CLA was analyzed according to the DPPH, ABTS and hydroxyl radical-scavenging activity, and also analyzed to enzyme inhibitory activity. Antioxidant activity was 83.9%, 89.3% and 81.6% in concentration of 1 mg/g of fermented soybean included CLA, respectively, but enzyme inhibitory activity against a-glucosidase and pancratic lipase was weak as 18.7% and 14.9%, respectively. The present finding may be used as a functional food or a medicine.

B. study on commercialization of CLA-containing soy-fermentation products using the fermentation of fungi

1. Production of extracts and dietary feeds from CLA-containing soy-fermentation using lactic acid bacteria (LAB)

We compared the changes of conjugated linoleic acid (CLA) and isoflavone-aglycone contents and biological effects, α-glucosidase and pancreatic lipase inhibition activities of soybean powder milk of Korean soybean cultivars, such as Saedanbaek, Daewon, Wooram and USA soybean by a potential probiotic Lactobacillus plantarum P1201. In the case of soybean powder milk of Saedanbaek cultivar, after 72 h of fermentation, the pH decreased from 6.0 of initial fermentation to 3.98, while the acidity, viable cell numbers and protein contents increased to 1.25%, 9.43 log cfu/g and 254.92 mg/g, respectively. A 1 mg/ml of fermented Saedanbaek from soybean powder milk were ABTS (79.77%) and hydroxyl radical scavenging activities (52.26%). In other hand, α-glucosidase (78.72%) and pancreatic lipase inhibition activities (18.51%) results were the highest in fermented Wooram soybean cultivar compared with the other samples examined. Importantly, the total phenolic, flavonoids, CLA, and isoflavone-aglycone contents increased, but glycoside and malonyl-glycosides contents decreased. The levels of daidzein, glycitein, and genistein were obtained 213.07 μg/g, 91.88 μg/g and 109.97 μg/g in fermented wooram cultivar, respectively. In particular, the contents of cis-9, trans—11 CLA and trans-10, cis-12 CLA were 1.04 mg/g and 0.11 mg/g, respectively.

Fermented soy-powder milk (FSPM) was made by fermentation with Lactobacillus plantarum P1201. The differences in the physicochemical properties, bioactive compounds, and biological activities between unfermented soy-powder milk (UFSPM) and FSPM were investigated. FSPM had a higher level of conjugated linoleic acid (CLA) and isoflavone aglycones compared with UFSPM. Both FSPM and UFSPM showed dose-dependent inhibitory activities for α-glucosidase, α-amylase, and pancreatic lipase.

2. Selection of the fungi with produced higher CLA and soybean cultivars

We investigated the carbohydrate, pancreatic lipase inhibition activity and isoflavone contents of CLA-containing fermented soybean by Tricholoma matsutake, Polyozellus multiplex, and Grifola frondosa mycelium of mushroom. The fermented soybean by P. multiplex CLA content were highest showed the cis-9, trans-11(42.94 μg/g), trans-10, cis-12 (3.27 μg/g) and total CLA (46.21 μg/g) respectively. The highest levels of linoleic acid (C18:2C) were present at concentrations of 3,174.4 mg/100 g (T. matsutake) and 1,348.9 mg/100 g (G. frondosa) respectively. In consequence, the levels of total phenolic and isoflavone-aglycone contents, ABTS, DPPH and ∙OH radical scavenging activities, and FRAP assay values increased, while isofalvoneglycoside contents decreased during the fermentation by mycelium of mushroom. In addition, the α-glucosidase, α-amylase and pancreatic lipase inhibition activities of fermented soybean (T. matsutake) were 93.00%, 30.21% and 35.00%, respectively.

The changes in conjugated linoleic acid (CLA) and isoflavone contents and biological activities during the solid-state fermentation of four soybean cultivars by Polyozellus multiplex were investigated. During solid-state fermentation of different soybean cultivars, the total phenolic and isoflavone-aglycone (daidzein, glycitein and genistein) contents increased, while radical scavenging activities and α-glucosidase inhibition activity, but glycoside and malonyl-glycosides (daidzin, glycitin, genistin, malonyldaidzin, malonylglycitin, and malonylgenistin) contents decreased. The highest levels of daidzein, glycitein, and genistein were obtained 429.37, 35.75, and 285.56 μg/g in the finial fermentation day (8 day) with Daepung cultivar. In consequence, the results of DPPH and ABTS radical scavenging activities and α-glucosidase inhibition activity were the highest in Daepung cultivar than other soybean cultivars. The fermented soybeans showed the higher CLA contents and biological activities compared to any other unfermented soybeans examined. In particular, the levels of cis-9, trans-11, trans-10, cis-12, and total CLA were detected to be 47.25 μg/g, 4.63 μg/g, and 51.88 μg/g in the finial fermentation day (8 day) with Daepung cultivar.

3. Optimal fermentative condition of produced CLA from soybean using fungi

The changes in β-glucosidase activity, total phenolic, CLA, and isoflavone contents, and biological activities during the solid-state fermentation of soaking soybean (SSB) and germinated soybean (GSB) by Polyozellus multiplex were investigated. The levels of β-glucosidase activity and total phenolic, CLA and isoflavone-aglycone contents increased, while antioxidant activity (DPPH, ABTS, and hydroxyl radical scavenging activities and FRAP assays) and α-glucosidase inhibition activity increased, but the isoflavone-glycoside and –malonyglycoside contents decreased after the solid-state fermentation of soybean. In particular, the germinated soybean fermented at 25°C for 12 days displayed the highest antioxidant activities, compared to those of the soaking soybean and this of soybean fermented. Also, the highest levels of daidzein, glycitein, and genistein were present at concentrations of 584.24 μg/g, 178.34 μg/g, and 370.47 μg/g at the finial fermentation day (8 days) of the germinated soybean fermentation. In other hand, the highest levels of cis-9, trans-11 CLA, trans-10, cis-12 CLA, and total CLA were present at concentrations of 50,35 μg/g, 4.87 μg/g, and 55.22 μg/g at the 12 days of the non-germinated soybean fermentation, respectively.

The comparison of quality characteristics, phytochemical contents (total phenolic and isoflavone), and biological activities during the solid-state fermentation of soybean meat by mycelia of pine mushroom were investigated. The fermented soy meat showed the texture of hardness, springiness, chewiness, and gumminess were 114.2, 3.98, 212.20, and 54.77 respectively.

Color (L, A, and B value) characteristics was higher in steamed soy meat more than fermented soy meat. Meanwhile, cis-9, trans-11(0.91 μg/g) and trans-10, cis-12(0.18 μg/g) isomer contents increased of fermented soy meat but not detected of steamed soy meat. The total phenolic (630.42 μ g/g), flavonoid (37.30 μg/g) and isoflavone-aglycone contents (154.23 μg/g) were highest in the fermented soy meat. As a results, free radical scavenging activities (DPPH, ABTS, ⦁OH, and FRAP) in fermented soy meat were show to be high 61.70%, 87.56%, 66.68%, and 2.017, respectively.

4. Analysis of microflora according to CLA-containing soy-powder yogurt diets

The predominant species was Proteobacteria, high-fat diet (HFD) 75.0% and high-fat diet with extracts (HFD + SPY) 64.3%, respectively.

C. Study on effect of body fat reduction of CLA containing fermented soybean

1. study of the anti-obesity effect of Fermented soy powder milk(FSPM) by using cell culture system and in vivo model FSPM inhibited the lipid accumulation in the adipocyte(3T3-L1) and the differentiation of adipocyte. The total amount of lipid in adipocyte was also decreased by FSPM. LPL, C/EBP-α, adiponectin, aP2, FAS and ACC(which are related with lipid accumulation and adipocyte differentiation) mRNA were decreased by FSPM. Un-fermented SPM(UFSPM) did not have these functions, even more some adipocyte trophic factor was increased by UFSPM treating.

The High fat diet(HFD) fed mice with FSPM had body weight less than 29% compared to HFD only mice group. HFD mice had liver weight by double compared to normal diet group(ND), FSPM decresed the change by 33%. By blood analysis, we could know, FSPM decreased blood sugar level, total cholesterol and LDL significantly compared to HFD. The fatty liver change was also partially blocked by FSPM by decreasing the expression level of PPARγ, TNF-α and IL1β.

2. study of the anti-obesity effect of Fermented soy powder milk(FSPM) on HFD induced fat mice

After inducing fatty boy by feeding HFD for 5weeks, FSPM decreased the body weight, liver weight and degree of fatty liver change. We harvested the damaged liver tissue and analyzed the enzymatic change by RT-PCR, FSPM decreased the mRNA level of SREBP-1c, aP2, ACC, FAS, PPARγ and LPL.

To identify the key substance of this effect, we designed another experiment, CLA only group VS FSPM group, total amount of CLA was adjusted equally in both group. CLA group and FSPM group had decreased expression level of PPARγ, FSPM group had lower level of aP2 expression by CLA group. Other analyses were not significant.

(출처 : Summary p.10)

목차 Contents

• 표지 ... 1제출문 ... 2보고서 요약서 ... 3국문요약문 ... 4Summary ... 10목차 ... 16제 1 장 연구 개발 과제의 개요 ... 17 1절 연구 개발 목적 ... 17 2절 연구 개발의 필요성 ... 17 3절 연구 개발 범위 ... 23제 2 장 국내외 기술개발 현황 ... 24 1절 국내 연구 현황 ... 24 2절 국외 연구 현황 ... 25제 3 장 연구 수행 내용 및 결과 ... 27 1절 연구개발 방법 및 내용 ... 27 2절 연구개발 결과 및 고찰 ... 48제 4 장 연구개발목표 달성도 및 대외기여도 ... 191 1절 목표대비 달성도 ... 191 2절 정량적 성과(논문게재, 특허출원, 기타)를 기술 ... 192제 5 장 연구 결과의 활용 계획 ... 193 1절 활용방안 및 추가 연구 ... 193 2절 추가적인 논문게재, 산업재산권 출원 등 ... 193제 6 장 연구 과정에서 수집한 해외 과학 기술 정보 ... 194 1절 국내 건강기능성식품 최근 동향 ... 194 2절 인간의 체질 및 장내미생물과의 상관성 연구 ... 194제 7 장 연구 개발 결과의 보안 등급 ... 195제 8 장 국가과학기술지식정보서비스에 등록한 연구시설·장비 현황 ... 196제 9 장 연구개발과제 수행에 따른 연구실 등의 안전조치 이행실적 ... 197제 10 장 연구개발과제의 대표적 연구실적 ... 199제 11 장 기타사항 ... 200제 12 장 참고문헌 ... 201끝페이지 ... 204