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Kafe 바로가기주관연구기관 | 세종대학교 Sejone university |
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연구책임자 | 이수용 |
참여연구자 | 유상호 , 김유리 , 정상원 , 홍재희 |
보고서유형 | 최종보고서 |
발행국가 | 대한민국 |
언어 | 한국어 |
발행년월 | 2017-04 |
주관부처 | 농림축산식품부 Ministry of Agriculture, Food and Rural Affairs(MAFRA) |
등록번호 | TRKO201800009771 |
DB 구축일자 | 2018-05-26 |
키워드 | 수국.고감미 감미료.설탕 보완재.자일로바이오스.필로둘신.Hydrangea macrophylla.Sweetener.Sugar complement.Xylobiose.Phyllodulcin. |
DOI | https://doi.org/10.23000/TRKO201800009771 |
본 연구 사업에서는 설탕 보완재로써 xylobiose 당류의 생촉매적 합성과 함께 천연고감미소재인 phyllodulcin의 추출, 분리를 통한 산업규모의 생산 가능성을 타진하여 새로운 식품소재로써의 활용성을 타진한 연구로 연구개발결과가 매우 우수하며, 후속연구를 통한 식품, 화장품분야의 핵심소재로써의 가능성이 매우 높음. 연구수행이 성실도가 매우 높으며, 국내기반 핵심소재개발기술의 배타적 권리 확보 역시 잘 진행된 것으로 판단됨.
(출처 : 요약서 3p)
Purpose&Contents
◯ Objective and Necessity of Research and Development To establish an efficient production process of xylobiose by biocatalyst system, and to apply an purification and crystallization technology for development xylobisose containing sugar complementary material. In addition, we d
Purpose&Contents
◯ Objective and Necessity of Research and Development To establish an efficient production process of xylobiose by biocatalyst system, and to apply an purification and crystallization technology for development xylobisose containing sugar complementary material. In addition, we developed efficient biocatalytic pretreatment, extraction and purification of phyllodulcin which is a high intensity sweetener from hydrangea leaves. As a result, we developed a new sweetener with proven healthy functionality of anti-diabetic and anti-obesity by quality improvement of phyllodulcin.
◯ Content and Scope of Research and Development
· Development of biosynthetic xylobiose for sugar supplement
· Development technology of purification and crystallization on xylobiose
· Elucidation of inhibitory effect of blood glucose and prebiotic effect by xylobiose
· Development for efficient extraction and purification of phyllodulcin from hydrangea leaves
· Development of fermentation technology for increasing phyllodulcin content from hydrangea leaves
· Improvement solubility and sweetness of phyllodulcin by catalystic and excipient technology
· The anti-diabetic and anti-obesity effects of xylobiose, and the anti-diabetic and anti-obesity effects of phyllodulcin
· Comprehensive sweetness characteristics of phyllodulcin and xylobiose and their sweetness stability in a food system
Results
◯ Optimized production of phyllodulcin from hydrangea leaves and elucidation of its properties
· Phyllodulcin and hydrangenol were identified from the extract of hydrangea leaves using HPLC-MS.
· Optimum one-pot massive bioconversion and extraction could be utilized industrially for phyllodulcin production in an easy, quick and cost-effective way.
· Mixed-bed ion exchangers removed impurities and colorants efficiently, and the resulting solution displayed over 90% of very high recovery from the phyllodulcin extract.
· By utilizing preparative HPLC, highly pure phyllodulcin was finally produced with 98.3% of purity and 2% yield of which the productivity can be obtained from scale-up process.
· By utilizing crystallization process, highly pure phyllodulcin was finally produced with 97.3% of purity and 1.6% yield of which the productivity can be obtained from scale-up process.
· Crystallization process of phyllodulcin production was cost-effective more than purification of preparative HPLC.
· The inclusion complexation of phyllodulcin with β-cyclodextrin was successfully achieved, which was confirmed by various instrumental analyses such as UV/Vis spectrophotometer, DSC, FT-IR and 1H-NMR.
· The solubility and antioxidant activity of inclusion complex between phyllodulcin and β-CD was remarkably higher than that of phylldouclin.
◯ Development of biosynthetic xylobiose for sugar supplement
· Recombinant GRAS strains (Yeast, Bacillus) were constructed by β-xylosidase (XynB) from B.pumilus which had high biosynthesis efficiency.
· To optimize the large scale culture process of β-xylosidase production, the cultivation was successfully carried out in 30 L reactor system (cell mass > 8.4 g DCW/L).
· According to the result of optimization of purification process, purification yield of β-xylosidase was more than 80%.
· β-xylosidase from Y01 strain was used to optimize the xylobiose biosynthesis process. As a result, the maximum xylobiose conversion ratio was 17.6% when the reaction was conducted at 95% (w / v) xylose, 10 U / mL, temperature 50℃.
· Simulated Moving Bed (SMB) was applied to establish large scale purification system and final purity of biosynthetic xylobiose was 98.5%.
· Xylobiose showed rat sucrase inhibition potency and prebiotic effects. XylobioSugar< provisional name > reduced blood glucose response and it is a low-GI food.
◯ Comprehensive sweetness characteristics of phyllodulcin and xylobiose and their sweetness stability in a food system
· Sucrose complement, Xylobiose was added to sugar and xylobiose-sugar mixture was evaluated for mixing suitability with sugar, separation stability, and caking characteristics.
XylobioSugar< provisional name > was developed and used it to make stick sugar and coffeemix prototype.
· The methodology to elucidate comprehensive sensory profiles of potential natural alternative sweeteners was developed using xylooligosaccharide(XOS) and luo han guo extract(LHGE) as model sweeteners.
· The sweetness potency of phyllodulcin and XB, relative to that of sucrose, was investigated using 2-alternative forced choice (AFC) test. Phyllodulcin solution was prepared by dissolving it in 1% ethanol (v/v) at 50oC. For phyllodulcin, relative sweetness(RS) to that of 3% sucrose in 1% ethanol solution was determined. XB was prepared as aqueous solution and RS to 5% sucrose was determined. RS of phyllodulcin and XB was 434.8, and 0.34, respectively.
· The sensory profile of phyllodulcin and XB was elucidated based on descriptive analysis (DA) using 10 trained panelists.
Phyllodulcin was characterized by its bitterness, alcohol flavor, cooling sensation, and astringent aftertaste, but the persistence and onset of the sweetness of phyllodulcin were similar to those of sucrose. XB was distinguished from other sweeteners by its yellowness, corn aroma, corn flavor, and nurungji candy flavor.
Persistence of sweetness of XB seems to be similar to that of sucrose, but its onset of sweetness was slower than that of sucrose. When XB was mixed with sucrose at the ratio of 7:93, the mixture exhibited similar sensory quality as that of sucrose, making it a useful sucrose complement.
· RS of phyllodulcin to that of 15.3% sucrose was identified in a cookie system using 2-AFC. Cookie model samples were formulated by replacing sucrose with phyllodulcin. Because intense bitterness of phyllodulcin suppressed sweetness at high concentration ranges, phyllodulcin could not produce equi-intense sweeteness to 15.3% sucrose. When phyllodulcin was mixed with sucrose at the ratio of 1:1, its RS to 7.7% sucrose was identified as 333.3.
· Sensory characterisitcs of cookies containing phyllodulcin and xylobiose were analyzed using QDA. Phyllodulcin replaced 50% and 100% of sugar in the cookie formulation, respectively.
Xylobiose mixed with sugar at the ratio of 7:93 was added to the cookie in place of sugar. 100% phyllodulcin cookie (PHY100) was significantly less sweet, and had stronger umami, hardness, hardness of fragments, fracturability, and lingering sensations of umami, acridness, and bitterness compared to 100% sugar cookie(SUC). Sweetness of 50% phyllodulcin cookie (PHY50) was not significantly different from that of SUC, and also PHY50 exhibited very weak umami, bitterness, and acridness.
Xylobiose-sugar cookie (XOBS) was characterized by brown color, aroma and flavor of roasted soy flour, and heterogenity of particles.
· Consumer liking of cookies containing alternative sweeteners was assessed both in blind and informed condition to understand the effect of information on consumer liking. Overall liking scores of all cookies were increased by increased by information provided, but information had only limited effect on liking of PHY100 that was least liked in blind condition, because its liking did not become significantly greater than those of other cookies.
Consumers liked SUC most, but there was not significant difference in liking between SUC and PHY50 and XOBS.
◯ The anti-diabetic and anti-obesity effects of xylobiose, and the anti-diabetic and anti-obesity effects of phyllodulcin.
· In vitro and in vivo methodology system to examine the anti-diabetic and anti-obesity effect of xylobiose and phyllodulcin was established using D-xylose and stevioside.
· Xylobiose replaced by glucose in media increased glucose uptake in skeletal muscle cell and insulin secretion in pancreatic β-cells.
· Xylobiose supplementation decreased the level of fasting blood glucose, glucose tolerance and regulated plasma lipid profiles such as TG, TC, and LDL compared to diabetic control in db/db mouse, respectively.
· Xylobiose supplementation decreased fasting blood glucose, plasma lipid profiles such as TG, TC, and LDL, and also the weight of subcutaneous fat and mesenteric fat.
· The size and number of hepatic lipid droplet were decreased by xylobiose supplementation. Hepatic lipogenesis markers such as C/EBPα, PPARγ, SREBP-1c were down-regulated in xylobiose supplemented group compared to obesity control.
· Glucose uptake in skeletal muscle cell and insulin secretion in pancreatic β-cell were increased by 1, 3, or 10 uM phylloduclin treatment, respectively.
· Phyllodulcin supplementation decreased water and food intake, fasting blood glucose and glucose tolerance in db/db mouse.
· Phyllodulcin supplementation decreased the size of lipid droplet in the liver tissues and decreased gluconeogenesis marker such as PEPCK expression in the liver and increased glucose uptake marker, GLUT4 expression in the muscle of db/db mice.
· Phylloduclin treatment decreased lipid accumulation in differentiated 3T3 L1 cells and down-regulated mRNA levels of lipogenesis and inflammation markers compared to glucose 20mM. The protein expressions of c-Raf signaling, including c-Raf, p-Akt, p-mTOR were decreased in phyllodulcin treatment, respectively.
· Phyllodulcin supplemenation decreased fasting blood glucose, plasma lipid profile such as TG, and the weight of subcutaneous fat. In addition, lipogenesis markers, including C/EBPα, PPARγ, SREBP-1c in subcutaneous fat were lower in phyllodulcin supplementation groups compared to obesity control group.
Expected Contribution
◯ Research achievements
· Paper publication 10 (SCI 5)
· Academic achievements (proceeding, poster etc.) 15
· Graduate students education 10
· Patents (application 4, registration 1)
· Prototype 3
◯ Application of research achievements
· Patent application for ‘Scale-up production of phyllodulcin, a potential high intensity sweetener, from hydrangea leaves by crystallization method’ is planned for the first half of 2017.
· Patent application for ‘Phyllodulcin quality improvement of inclusion complexation between phyllodulcin and beta-cyclodextrin’ is planned for the first half of 2017.
· Paper publication for ‘Effects of xylobiose-sugar mixture on defecation frequency and symptoms’ is planned for the first half of 2017.
· Paper publication of ‘Relative sweetness and sweetness quality of Xylobiose’ is planned in April 2017.
· Review of technology transfer for ‘Production process technology of phyllodulcin‘ or ‘Production process technology of biosynthetic xylobiose for sugar complement’
· This study has established the methodologies to evaluate relative sweetness, sweetness quality, and temporal sweetness profile of natural alternative sweeteners that were characterized by strong bitter off-flavor. Also this study has developed the method to solubilize phyllodulcin in a model solution using alcohol and heat, and the plain sugar cookie system that can be used as a food model system. The methods and model systems developed in this study can contribute to future research of natural alternative sweetners with low solubility.
· This study has identified relative sweetness, sweetness quality, and temporal sweetness profiles of phyllodulcin and xylobiose that had not been much studied. The outcomes of this study can provide academia and industry with a useful guideline for application of the nutural alterantive sweeteners to foods and beverages.
· By investigating consumer liking and perception of the cookie model system which phyllodulcin and xylobiose are applied to, consumer liking and major drivers of liking have been identified.
This result will contribute to future studies to improve sensory quality and functionality of natural alternative sweeteners.
· This study investigated the potential effect of information about functionality and health-promoting effect of alternative sweeteners on consumer liking and purchase intent. The outcomes of this study can help food industries to establish a marketing strategy for commercialization of alternative natural sweeteners.
· The methodology for evaluating in vitro and in vivo anti-diabetic and anti-obesity effect of high intensity sweetener and a sugar complement was established and would be contributed for the researches in this area.
· Suggest the scientific evidence for effects of xylobiose and phyllodulcin on anti-diabetic and anti-obesity.
· Establish the various R&D foundation through technical development and professional manpower training.
· Expect strengthen overseas export and import substitution effect of high-sweetness sweeteners.
(출처 : SUMMARY 10p)
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