초록 ▼

○ 연구결과
① 감귤류로부터 NHDC 생산 원료물질의 함량 분석 및 함량 증진 기술 개발
- 감귤류로부터 품종 및 숙성 시기에 따른 NHDC 생산 원료물질의 함량 분석
- 효소적 처리법에 의한 naringenin, hesperidin 10 배 이상의 함량 증진 기술 확보
② 감귤과피로부터 NHDC 생산 원료인 neohesperidin 추출 공정 개발
- 반응표면분석법(RSM)을 이용한 추출조건 확립
③ Neohesperidin 생전환용 효소 생산 균주의 탐색 및 배양 조건의 확립
- Neohes

○ 연구결과
① 감귤류로부터 NHDC 생산 원료물질의 함량 분석 및 함량 증진 기술 개발
- 감귤류로부터 품종 및 숙성 시기에 따른 NHDC 생산 원료물질의 함량 분석
- 효소적 처리법에 의한 naringenin, hesperidin 10 배 이상의 함량 증진 기술 확보
② 감귤과피로부터 NHDC 생산 원료인 neohesperidin 추출 공정 개발
- 반응표면분석법(RSM)을 이용한 추출조건 확립
③ Neohesperidin 생전환용 효소 생산 균주의 탐색 및 배양 조건의 확립
- Neohesperidin 생전환용 효소 생산 균주 3종 확보
- 효소의 특성 분석
- 생전환용 효소 분비 미생물의 동정 및 대량배양 조건 확립
④ Neoheperidin으로부터 NHDC 제조 공정 개발
- 유기합성법에 의한 NHDC 수율 95% 이상 생산 기술 확보
- 6-sigma 기법을 이용한 neohesperidin 생전환용 효소 생산 균주의 고농도 배양 조건 확립
- 6-sigma 기법을 이용한neohesperidin 추출 조건의 확립

Abstract ▼

Ⅱ. Synopsis of research and development subject
1. Object of research
The present report offers an analysis of the useful compounds from citrus peel waste and develope the process of the preparation of neohesperedin dihydrochalcone from citrus peel waste.
2. Necessity of research
Categor

Ⅱ. Synopsis of research and development subject
1. Object of research
The present report offers an analysis of the useful compounds from citrus peel waste and develope the process of the preparation of neohesperedin dihydrochalcone from citrus peel waste.
2. Necessity of research
Categorically, high intensity sweeteners fall into two classes: artificial sweeteners and natural sweeteners. Currently, the share of artificial sweeteners in the high intensity sweeteners market is the largest, - major reason behind the popularity is their low content of calories and a taste which is similar to that of sugar. Low calorie sweeteners do not contain fermentable carbohydrates, and eventually do not produce acids by oral bacteria, and therefore, they prevent dental carries. Diabetic people also prefer low calorie sweeteners as it provides them the same sweet taste of sugar without side effects. Their low calorie property does not add any calories to the diet of diabetic patients and thus helps in weight management, which, in turn, helps in preventing many diseases like obesity and heart related diseases.
A series of factors have added to the growth of high intensity sweeteners (HIS) over the past few years, with the major of them being the rising health concerns and increasing demand of these sweeteners in the food and beverage industry worldwide. Aspartame is the largest artificial sweetener among others in terms of its market share, which is followed by sucralose and saccharin. As an intense sweetener, aspartame is mostly demanded by the food and beverage industry. Globally, rising population of people with weight related diseases, like diabetes and obesity, the choice of consumers is shifting towards artificial and natural sweeteners sugar.
NHDC has been a commercial product for many years; however, production is limited by the availability of the substrate neohesperidin, which accumulates to significant levels only in inedible citrus species such as Citrus aurantium (bitter orange) and other citrus hybrids that are especially grown for this purpose
Ⅲ. Contents and results of the research
We propose a process to convert hesperidin, a tasteless flavonoid extracted from orange peels that are abundant byproducts of the vast orange juice industry, into neohesperidin using three steps: (i) extraction of hesperidin from orange peels, (ii) biotransformation of hesperidin hydrolysis products into neohesperidin, and (iii) organic synthesis of NHDC from neohesperidin. We overcame the current technological bottleneck in biotransformation of hesperidin hydrolysis products into neohesperidin using flavanone-7-O-glucoside-2-O-rhamnosyltransferaseproduced by Steptomyces sp.. A small-scale production experiment established the feasibility of the proposed process.
1. Analysis of useful compounds from citrus peel waste
From mid January to the end of February 2007, citrus fruits were collected from 12 different small farms in Jeju island. Up to 1 kg of fully ripe, 10−11 months old citrus fruits (in-seasonal produced) were available from each farm. Conventional grown citrus fruits were used as a control because these are cultivated without a guideline on the usage of fertilizers and pesticides. Hesperidin was the predominant flavanone concentration, being much higher than those of other flavonoids. In the citrus peel, two flvonoids, including hesperidin, neohesperidin contents was the highest during early August.
2. Conversion of hesperidin into neohesperidin
We screened microorganisms for conversion of hesperidin into neohesperidin, and then selected Streptomyces yatensis, Streptomyces misionensis, Streptomyces hygroscopicus subsp. glebosus having glycosyltranferase activity. A small-scale production experiment was conducted to establish the feasibility of the process presented above for conversion of orange peel hesperidin into neohesperidin.
IⅤ. Practical application of the research results
NHDC, Low calorie sweeteners do not contain fermentable carbohydrates, and eventually do not produce acids by oral bacteria, and therefore, they prevent dental carries.
Diabetic people also prefer low calorie sweeteners as it provides them the same sweet taste of sugar without side effects. Their low calorie property does not add any calories to the diet of diabetic patients and thus helps in weight management, which, in turn, helps in preventing many diseases like obesity and heart related diseases. It also discusses major market trends, growth drivers, opportunities available for the players and major challenges faced by the sweetener industry.
While the data show our approach to NHDC production is feasible at the laboratory level, much research is needed to advance toward application. Three apparently correlated drawbacks that need to be worked out are (i) inconsistent levels of neohesperidin production and (ii) a significant proportion of the hydrolyzed hesperidin substrate is converted into a diglucoside at the expense of the desired neohesperidin product, (iii) a increse of the conversion rate neohesperidin into NHDC

목차 Contents

• 표지 ... 1
• 제 출 문 ... 2
• 요 약 문 ... 3
• SUMMARY ... 7
• CONTENTS ... 10
• 목 차 ... 12
• 제 1 장 연구개발과제의 개요 ... 14
• 제 1절 연구 개발의 목적 ... 14
• 1. 연구 목적 ... 14
• 2. 감미료 개발의 필요성 ... 15
• 제 2절 연구 개발의 필요성 ... 24
• 1. 기술적 측면 ... 24
• 2. 경제·산업적 측면 ... 24
• 3. 사회·문화적 측면 ... 25
• 제 2 장 국내외 기술개발 현황 ... 27
• 제 3 장 연구개발 수행 내용 및 결과 ... 29
• 제 1절 감귤가공 부산물의 물질의 분석 ... 29
• 1. 감귤품종 및 숙성시기에 따른 원료 물질 함량 분석 ... 29
• 2. 숙성시기에 따른 원료 물질 함량 분석 ... 39
• 제 2절 원료물질의 추출 및 함량 증진 ... 41
• 1. 산업용 효소 처리에 의한 원료물질의 함량 변화 ... 41
• 2. 원료물질의 추출 공정 확립 ... 50
• 3. 반응표면분석법(Response surface methodology)을 이용한 추출 공정의 확립 ... 53
• 제 3절 당 전이효소를 이용한 neohesperidin의 생산 ... 59
• 1. 균주의 선발 ... 59
• 2. Glycosyl-group 전이 효소 생산 미생물의 순수분리 ... 61
• 2. Glycosyltranferase 효소의 분리 정제 및 특성 분석 ... 67
• 3. Streptomyces yatensis 대량생산 기술 개발 ... 72
• 제 4절 Neohespertin으로부터 NHDC 생산 공정의 개발 ... 80
• 1. 유기합성법에 의한 NHDC 생산 공정 개발 ... 80
• 2. NHDC 시제품 생산 ... 83
• 제 5절 바이오공정개선을 위한 6-Sigma의 융합연구 ... 84
• 1. 6-Sigma 기법을 이용한 초임계 추출 조건의 최적화 ... 84
• 2. 6-Sigma 기법을 이용한 S. yatensis의 배양 조건의 최적화 ... 97
• 제 4 장 목표달성도 및 관련분야에의 기여도 ... 105
• 제 1절 목표달성도 ... 105
• 제 2절 관련분야에 대한 기여도 ... 106
• 제 5 장 연구개발 성과 및 성과활용 계획 ... 107
• 제 6 장 참고문헌 ... 108
• 끝페이지 ... 111