KR20240061963A - Development of novel slowly digestible carbohydrates for balanced postprandial glycemic response and regulation of energy metabolism - Google Patents
Development of novel slowly digestible carbohydrates for balanced postprandial glycemic response and regulation of energy metabolism Download PDFInfo
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- 150000001720 carbohydrates Chemical class 0.000 title abstract description 11
- 238000011161 development Methods 0.000 title abstract description 4
- 230000000291 postprandial effect Effects 0.000 title abstract description 4
- 230000037149 energy metabolism Effects 0.000 title abstract description 3
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/20—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents
- A23L29/269—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents of microbial origin, e.g. xanthan or dextran
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/328—Foods, ingredients or supplements having a functional effect on health having effect on glycaemic control and diabetes
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2250/00—Food ingredients
- A23V2250/50—Polysaccharides, gums
- A23V2250/502—Gums
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- General Chemical & Material Sciences (AREA)
- Dispersion Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Polymers & Plastics (AREA)
- Microbiology (AREA)
- Food Science & Technology (AREA)
- Nutrition Science (AREA)
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- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
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Abstract
본 발명은 식후 혈중 농도 균형 및 생체내 에너지 대사 조절을 위한 신개념의 지소화성 탄수화물 소재 개발에 관한 것으로, 웨이셀라 콘푸사 (Weissella confusa) DSM 20196 및 웨이셀라 사이베리아 (Weissella cibaria) DSM 14295 균주를 이용하여 각각 약 1.03×108 g/mol, 약 6.47×107 g/mol의 분자량을 가지며, α-1,6 결합을 90% 이상 가지는 지소화성 알파 글루칸의 제조방법 및 식품산업 적용 방안을 제공한다.The present invention relates to the development of a new concept of lipodigestible carbohydrate material for postprandial blood concentration balance and in vivo energy metabolism control, using Weissella confusa DSM 20196 and Weissella cibaria DSM 14295 strains. Thus, it provides a manufacturing method and food industry application plan for lipotropic alpha glucan, which has a molecular weight of about 1.03 .
Description
본 발명은 식후 혈중 농도 균형 및 생체내 에너지 대사 조절을 위한 신개념의 지소화성 탄수화물 소재 개발에 관한 것이다. 더욱 상세하게는, 웨이셀라 콘푸사 (Weissella confusa) DSM 20196 와 웨이셀라 사이베리아 (Weissella cibaria) DSM 14295 균주들을 이용하여 각각 약 1.03×108 g/mol, 약 6.47×107 g/mol의 분자량을 가지며, α-1,6 결합을 90% 이상 가지는 지소화성 알파 글루칸의 제조방법 및 식품산업 적용 방안을 제공하고자 한 것이다.The present invention relates to the development of a new concept of lipodigestible carbohydrate material for postprandial blood concentration balance and control of in vivo energy metabolism. More specifically, molecular weights of approximately 1.03 × 10 8 g/mol and approximately 6.47 × 10 7 g/mol were obtained using Weissella confusa DSM 20196 and Weissella cibaria DSM 14295 strains, respectively The aim is to provide a manufacturing method and food industry application method for lipodigestible alpha glucan having more than 90% of α-1,6 bonds.
현재 우리나라를 포함한 주요 선진국의 비만 인구의 비율이 급속도로 증가하는 가운데, 세계 보건 기구에서는 비만 자체를 심각한 대사성 질환으로 정의하며 비만의 비율이 높아짐에 따라 제2형 당뇨병, 심장질환, 고혈압 또는 일부 암과 같은 만성질환의 비율이 같이 증가하여, 비만이 이와 같은 질환들의 주요 원인이 된다고 보고되었다. 대사성 질환으로 정의된 비만과 제2형 당뇨는 식이 탄수화물을 다량 섭취 시 혈중 포도당의 급격한 증감과 이에 따라 인슐린이 과다 분비됨으로 인해 인슐린 둔감성 등 생체내 항상성 조절의 불균형이 일어난다.Currently, the proportion of the obese population in major developed countries, including Korea, is rapidly increasing, and the World Health Organization defines obesity itself as a serious metabolic disease, and as the proportion of obesity increases, it leads to type 2 diabetes, heart disease, high blood pressure, and some cancers. It has been reported that the rate of chronic diseases such as is increasing, and obesity is the main cause of these diseases. Obesity and type 2 diabetes, defined as metabolic diseases, result in a rapid increase or decrease in blood glucose when consuming large amounts of dietary carbohydrates and subsequent excessive secretion of insulin, resulting in an imbalance in homeostatic control in the body, such as insulin insensitivity.
비만을 예방하기 위해서 가장 효율적인 에너지를 공급하는 탄수화물의 섭취를 줄이는 것은 오히려 영양학적 불균형을 초래하며 많은 부작용을 나타낼 수 있다. 정상인의 경우 포도당 대사 및 생체 내 영양분 공급 등을 유발하는 적절한 양의 탄수화물 섭취가 필요하기 때문에 최근에는 혈중 포도당의 급격한 증감 현상인 포도당 스파이크를 억제하고, 지속적으로 에너지를 공급하는 지소화성 탄수화물 소재가 생체 내 항상성 유지에 도움을 준다고 연구되었다. 이러한 지소화성 탄수화물은 인체의 장관 내에서 천천히 분해되어, 생체 내 지속적인 에너지 전달 및 인슐린의 급격한 농도 상승 억제를 통한 대사 과정의 항상성을 확보할 수 있는 신개념의 소재이다. In order to prevent obesity, reducing the intake of carbohydrates, which provide the most efficient energy source, can actually lead to nutritional imbalance and have many side effects. Since normal people need an appropriate amount of carbohydrate intake to induce glucose metabolism and nutrient supply in the body, recently, lipodigestible carbohydrate materials that suppress glucose spikes, which are a rapid increase or decrease in blood glucose, and provide continuous energy, have been used in the body. Studies have shown that it helps maintain homeostasis. These lipotropic carbohydrates are a new concept material that decomposes slowly in the human intestine and can ensure homeostasis of metabolic processes through continuous energy transfer in the living body and suppression of rapid concentration increases in insulin.
현재까지 지소화성 탄수화물의 경우 전분 유래의 지소화성전분(slowly digestible starches)에만 연구의 중점이 맞추어져 왔으며 이와 같이 아직까지 전분 외 다른 지소화성을 발휘하는 소재의 개발 및 그 제조방법에 대한 연구가 부족한 실정이다.To date, in the case of slowly digestible carbohydrates, research has focused only on slowly digestible starches derived from starch, and as such, there is still a lack of research on the development and manufacturing methods of materials that exhibit slow digestibility other than starch. This is the situation.
본 발명은 난소화성(in-digestible) 알파 글루칸으로 알려져 있는 웨이셀라(Weissella) 유산균으로부터 추출한 다당체(Exopolysaccharide, EPS)인 알파 글루칸이 섭취 시 식후 혈당의 급격한 증감을 억제하면서 에너지를 지속적으로 공급할 수 있는 천천히 소화되는 새로운 형태의 지소화성 소재임을 밝히고, 이와 같은 신소재의 제조방법 및 식품 산업으로의 적용 방법을 제공하고자 한다.The present invention provides that alpha glucan, a polysaccharide (EPS) extracted from Weissella lactic acid bacteria, known as indigestible alpha glucan, can continuously supply energy while suppressing rapid increases and decreases in postprandial blood sugar when ingested. We aim to reveal that it is a new type of slowly digestible material and provide methods for manufacturing this new material and applying it to the food industry.
본 발명은 배지와 증류수를 혼합한 후 자당과 맥아당을 첨가하여 배양액을 제조하는 단계 (a); 상기 단계 (a)에서 제조한 배양액에 웨이셀라 콘푸사 (Weissella confusa) 및 웨이셀라 사이베리아 (Weissella cibaria) 중 선택되는 어느 하나의 웨이셀라(Weissella) 속 균주를 접종하는 단계 (b); 상기 단계 (b)에서 웨이셀라(Weissella) 속 균주를 접종한 배양액을 원심분리하여 사균을 제거하는 단계 (c); 상기 단계 (c) 후, 배양액에 에탄올을 첨가하여 다당체를 침전시키는 단계 (d); 상기 단계 (d) 후, 상기 다당체가 침전된 배양액을 투석처리하여 다당체를 수득하는 단계 (e);를 포함하는 지소화성 알파글루칸의 제조방법을 제공한다.The present invention includes the step (a) of preparing a culture medium by mixing a medium and distilled water and then adding sucrose and maltose; (b) inoculating the culture medium prepared in step (a) with a strain of the Weissella genus selected from Weissella confusa and Weissella cibaria ; (c) removing dead cells by centrifuging the culture medium inoculated with the Weissella genus strain in step (b); After step (c), adding ethanol to the culture medium to precipitate the polysaccharide (d); After step (d), a step (e) of obtaining a polysaccharide by dialyzing the culture medium in which the polysaccharide has been precipitated is provided.
본 발명의 지소화성 알파글루칸의 제조방법에 있어서, 상기 투석처리는, MWCO (Molecular Weight Cut Off) 값이 3,500인 투석막(dialysis membrane)을 이용하여 수행하는 것을 특징으로 하는 것일 수 있다.In the method for producing lipotropic alpha-glucan of the present invention, the dialysis treatment may be performed using a dialysis membrane with a MWCO (Molecular Weight Cut Off) value of 3,500.
본 발명은 본 발명의 지소화성 알파글루칸의 제조방법에 의해 제조된 지소화성 알파글루칸을 포함하는 식품 조성물을 제공한다.The present invention provides a food composition containing lipotropic alpha-glucan prepared by the method for producing lipotropic alpha-glucan of the present invention.
본 발명의 지소화성 알파글루칸을 포함하는 식품 조성물에 있어서, 상기 식품은, 당뇨병 환자용 식이인 것을 특징으로 하는 것일 수 있다.In the food composition containing lipodigestible alpha-glucan of the present invention, the food may be characterized as being a diet for diabetic patients.
본 발명은 기존의 식이섬유/프리바이오틱 소재가 아닌 소장 전반에 걸쳐 지속적인 에너지 공급이 가능한 고점도의 지소화성 탄수화물 소재로써 생체 내 소화 모델에서 식후 혈당의 급격한 증가를 억제하며 동시에 에너지를 지속적으로 공급할 수 있고, 식이조절 호르몬의 분비를 자극시키는 지소화성 알파 글루칸을 제공할 수 있다.The present invention is not a conventional dietary fiber/prebiotic material, but a high-viscosity lipodigestible carbohydrate material that can continuously supply energy throughout the small intestine. It can suppress the rapid increase in blood sugar after a meal in an in vivo digestion model and simultaneously supply energy continuously. It can provide lipotropic alpha glucan that stimulates the secretion of diet-regulating hormones.
또한, 본 발명은 거대분자 크기의 고분자/고점도의 신개념 지소화성 다당체(EPS)로써 증점제로 활용할 수 있어, 소장 전반에 걸쳐 에너지를 공급할 수 있는 지소화성 탄수화물의 신개념이 접목된 점도 증진제로 당뇨 환자 또는 연하곤란 환자를 위한 환자식에 적용될 수 있는 지소화성 알파 글루칸을 제공할 수 있다.In addition, the present invention is a macromolecular-sized polymer/high viscosity new concept lipodigestible polysaccharide (EPS) that can be used as a thickener, and can be used as a viscosity enhancer incorporating the new concept of lipodigestible carbohydrates that can supply energy throughout the small intestine to patients with diabetes or Lipodigestible alpha glucans can be provided that can be applied to patient diets for patients with dysphagia.
도 1은 웨이셀라 콘푸사 (Weissella confusa) 및 웨이셀라 사이베리아 (Weissella cibaria) 두 종의 다당체를 산분해 처리 후 고성능 음이온 교환 크로마토그래피 (HPAEC)를 이용하여 각각의 구성당을 확인한 도면이다.
도 2는 생산된 2개의 다당체를 푸리에 변환 적외선 분광법(FT-IR)을 이용하여 구조를 확인한 도면이다 (A, Weissella confusa DSM 20196; B, Weissella cibaria DSM 14295; C, commercial dextran from Leuconostoc mesenteroides).
도 3은 생산된 2개의 다당체를 핵자기공명(NMR) 분광법 중 Heteronuclear Single Quantum Coherence (HSQC)를 이용하여 구조를 분석한 도면이다 (A, Weissella confusa DSM 20196 of HSQC NMR; B, Weissella cibaria DSM 14295 of HSQC NMR; C, commercial dextran from Leuconostoc mesenteroides of HSQC NMR).
도 4는 웨이셀라 콘푸사 (Weissella confusa) 및 웨이셀라 사이베리아 (Weissella cibaria)를 이용하여 포유류 유래의 소장 알파-글루코시다아제에 의한 분해속도(포도당의 유리 속도)를 갖는 것을 나타내는 도면이다.Figure 1 is a diagram showing the confirmation of each constituent sugar using high-performance anion exchange chromatography (HPAEC) after acid decomposition of polysaccharides of two species, Weissella confusa and Weissella cibaria .
Figure 2 is a diagram confirming the structure of the two produced polysaccharides using Fourier transform infrared spectroscopy (FT-IR) (A, Weissella confusa DSM 20196; B, Weissella cibaria DSM 14295; C, commercial dextran from Leuconostoc mesenteroides ).
Figure 3 is a diagram showing the structure analysis of the two produced polysaccharides using Heteronuclear Single Quantum Coherence (HSQC) among nuclear magnetic resonance (NMR) spectroscopy (A, Weissella confusa DSM 20196 of HSQC NMR; B, Weissella cibaria DSM 14295 of HSQC NMR; C, commercial dextran from Leuconostoc mesenteroides of HSQC NMR).
Figure 4 is a diagram showing the decomposition rate (glucose release rate) by mammalian small intestine alpha-glucosidase using Weissella confusa and Weissella cibaria .
본 발명은 생체 내 소화모델에서 천천히 소화됨에 따라 급격한 혈당의 증감을 억제하며, 지속적인 에너지를 공급할 수 있는 신개념의 지소화성 탄수화물 소재 제조방법 및 구조분석에 관한 것이다.The present invention relates to a method for manufacturing and structural analysis of a new concept of digestible carbohydrate material that can suppress rapid increases and decreases in blood sugar levels and provide continuous energy as it is digested slowly in an in vivo digestion model.
본 발명은 배지와 증류수를 혼합한 후 자당과 맥아당을 첨가하여 배양액을 제조하는 단계 (a); 상기 단계 (a)에서 제조한 배양액에 웨이셀라 콘푸사 (Weissella confusa) 및 웨이셀라 사이베리아 (Weissella cibaria) 중 선택되는 어느 하나의 웨이셀라(Weissella) 속 균주를 접종하는 단계 (b); 상기 단계 (b)에서 웨이셀라(Weissella) 속 균주를 접종한 배양액을 원심분리하여 사균을 제거하는 단계 (c); 상기 단계 (c) 후, 배양액에 에탄올을 첨가하여 다당체를 침전시키는 단계 (d); 상기 단계 (d) 후, 상기 다당체가 침전된 배양액을 투석처리하여 다당체를 수득하는 단계 (e);를 포함하는 지소화성 알파글루칸의 제조방법을 제공한다. 이때, 더욱 바람직하게는, 단계 (a)는 배지와 증류수를 혼합한 후 탄소원으로써 필요한 자당과 맥아당을 배지 내 1%가 되도록 첨가한 후 36~38℃에서 15~17 시간 배양시켜 배양액을 제조한다. 또한, 단계 (c)의 원심분리는 8,000 × g 에서 10~20 분 동안 진행하는 것이 바람직하며, 단계 (c)의 원심분리 후 배양액 2배 부피의 에탄올(99.9%)을 첨가하여 2~8℃에서 20~24 시간 용출하여 수용성 다당체를 수득하는 것이 바람직하다. 또한, 단계 (e)의 투석처리를 통해 단백질, 배지, 올리고당 등을제거하고, 동결건조하는 것이 바람직하다. 즉, 본 발명은 웨이셀라 콘푸사 (Weissella confusa) 및 웨이셀라 사이베리아 (Weissella cibaria) 중 선택되는 어느 하나의 웨이셀라(Weissella) 속 균주를 배지에 접종한 후 에탄올 침전을 통해 수용성인 다당체(EPS)를 얻은 다음 이를 투석처리를 통해 배지, 에탄올, 단백질 등을 제거하여 지소화성 알파글루칸인 다당체를 얻는 방법을 제공할 수 있다.The present invention includes the step (a) of preparing a culture medium by mixing a medium and distilled water and then adding sucrose and maltose; (b) inoculating the culture medium prepared in step (a) with a strain of the Weissella genus selected from Weissella confusa and Weissella cibaria ; (c) removing dead cells by centrifuging the culture medium inoculated with the Weissella genus strain in step (b); After step (c), adding ethanol to the culture medium to precipitate the polysaccharide (d); After step (d), a step (e) of obtaining a polysaccharide by dialyzing the culture medium in which the polysaccharide has been precipitated is provided. At this time, more preferably, in step (a), the culture medium is prepared by mixing the medium and distilled water, adding sucrose and maltose required as carbon sources to 1% of the medium, and culturing at 36-38°C for 15-17 hours. . In addition, the centrifugation in step (c) is preferably performed at 8,000 It is preferable to obtain a water-soluble polysaccharide by eluting for 20 to 24 hours. In addition, it is preferable to remove proteins, media, oligosaccharides, etc. through dialysis in step (e) and then freeze-dry. That is, the present invention inoculates a culture medium with a strain of the Weissella genus selected from Weissella confusa and Weissella cibaria and then inoculates water-soluble polysaccharide ( EPS ) through ethanol precipitation. ) can be obtained and then subjected to dialysis to remove the medium, ethanol, proteins, etc., thereby providing a method of obtaining polysaccharide, which is lipotropic alpha-glucan.
웨이셀라(Weissella)는 Lactobacillaceae(유산균과)에 속하는 그람 양성 박테리아의 속으로, 구형 또는 렌즈 모양의 세포에서 불규칙한 막대 형태까지 다양하다. 웨이셀라 콘푸사 (W.confusa) 및 웨이셀라 사이베리아 (W.cibaria)는 덱스트란 형태의 다당체를 생산하는 것으로 알려져 있다. 본 발명에서는 바람직하게 웨이셀라 콘푸사 (Weissella confusa) DSM 20196 및 웨이셀라 사이베리아 (Weissella cibaria) DSM 14295를 사용하는 것이 좋다. 상기 두 균주는 모두 생물자원센터 (Korean Collection for Type Cultures, KCTC)에서 분양받아 구입한 것이다. 웨이셀라 콘푸사 (Weissella confusa) DSM 20196은 KCTC3499로, 웨이셀라 사이베리아 (Weissella cibaria) DSM 14295는 KCTC3746으로 기탁되어 있는 균주이다. Weissella is a genus of Gram-positive bacteria belonging to the Lactobacillaceae family, with cells ranging from spherical or lens-shaped cells to irregular rod shapes. Weissella confusa ( W.confusa ) and Weissella cyberia ( W.cibaria ) are known to produce polysaccharides in the form of dextran. In the present invention , Weissella confusa DSM 20196 and Weissella cibaria DSM 14295 are preferably used. Both of the above strains were purchased from the Biological Resources Center (Korean Collection for Type Cultures, KCTC). Weissella confusa ( Weissella confusa ) DSM 20196 is a strain deposited as KCTC3499, and Weissella cibaria ( Weissella cibaria ) DSM 14295 is a strain deposited as KCTC3746.
한편, 본 발명의 지소화성 알파글루칸의 제조방법에 있어서, 상기 투석처리는 MWCO (Molecular Weight Cut Off) 값이 3,500인 투석막(dialysis membrane)을 이용하여 수행하는 것을 특징으로 하는 것일 수 있다. Meanwhile, in the method for producing lipotropic alpha-glucan of the present invention, the dialysis treatment may be performed using a dialysis membrane with a MWCO (Molecular Weight Cut Off) value of 3,500.
한편, 본 발명의 지소화성 알파글루칸의 제조방법에 있어서, 상기 다당체는, 곰팡이 유래의 글루코아밀라아제(glucoamylase)에 의해서는 분해되지 않고, 포유류 유래의 알파-글루코시다아제(α-glucosidase)에 의해서는 천천히 분해되는 것을 특징으로 하는 것일 수 있다. Meanwhile, in the method for producing lipotropic alpha-glucan of the present invention, the polysaccharide is, It may be characterized by not being decomposed by fungal-derived glucoamylase, but by being decomposed slowly by mammalian-derived alpha-glucosidase.
하기 실험에 의하면, 알파글루칸 형태의 미생물 유래 다당체(EPS)를 쥐 소장 유래의 알파-글루코시다아제 조효소에 의한 포도당 생산율을 통해 지소화성을 평가한 결과, 본 발명의 알파글루칸의 경우 기존의 난소화성 식이섬유 (dietary fibers)/프리바이오틱스(prebiotics)로 알려져 있던 미생물 유래 다당체(EPS)가 식이섬유 정량에 사용되는 아스퍼질러스(Aspergillus) 유래의 글루코아밀라아제(glucoamylase, AMG)에 의해서는 분해되지 않은 반면 포유류 유래 알파-글루코시다아제(α-glucosidase)에 의해 현저히 느린 속도로 분해되어 지속적으로 에너지를 공급할 수 있는 우수한 지소화성을 나타내는 점을 확인하였다.According to the following experiment, the indigestibility of microbial polysaccharide (EPS) in the form of alpha-glucan was evaluated through the glucose production rate by alpha-glucosidase coenzyme derived from rat small intestine. As a result, the alpha-glucan of the present invention had the existing indigestibility. Microorganism-derived polysaccharides (EPS), known as dietary fibers/prebiotics, are not decomposed by Aspergillus glucoamylase (AMG), which is used to quantify dietary fiber. On the other hand, it was confirmed that it is decomposed at a significantly slow rate by mammalian-derived alpha-glucosidase, showing excellent digestibility that can continuously supply energy.
또한, 하기 실험에 의하면 상기 다당체는 알파-1,6 결합을 90% 이상 가지는 덱스트란일 수 있는데, 웨이셀라 콘푸사 (Weissella confusa)를 이용한 경우에는 분자량이 약 1.03×108 g/mol, 웨이셀라 사이베리아 (Weissella cibaria)를 이용한 경우에는 분자량이 약 6.47×107 g/mol로 확인되었다. In addition, according to the following experiment, the polysaccharide may be dextran with more than 90% of alpha-1,6 bonds, and when Weissella confusa is used, the molecular weight is about 1.03 × 10 8 g/mol, Wei When Weissella cibaria was used, the molecular weight was confirmed to be about 6.47×10 7 g/mol.
한편, 본 발명은 본 발명의 방법에 의해 제조된 지소화성 알파글루칸을 포함하는 식품 조성물을 제공한다.Meanwhile, the present invention provides a food composition containing lipotropic alpha-glucan prepared by the method of the present invention.
한편, 본 발명의 지소화성 알파글루칸을 포함하는 식품 조성물에 있어서, 상기 식품은, 당뇨병 환자용 식이인 것을 특징으로 하는 것일 수 있다. 이와 같이, 본 발명의 지소화성 알파글루칸과 같은 지소화성 탄수화물 소재는 전분 외 다른 지소화성을 발휘하는 소재로 식후 혈당의 증감을 약화시킬 수 있으며 당뇨 환자를 위한 환자식에 적용될 수 있다.Meanwhile, in the food composition containing lipotropic alpha-glucan of the present invention, the food may be characterized as being a diet for diabetic patients. In this way, the lipotropic carbohydrate material, such as the lipotropic alpha-glucan of the present invention, is a lipotropic material other than starch and can attenuate the increase or decrease in blood sugar levels after a meal and can be applied to patient diets for diabetic patients.
이하, 본 발명의 내용에 대해 하기 실시예 및 실험예를 통하여 더욱 상세히 설명하고자 한다. 다만, 본 발명의 권리범위가 하기 실시예 및 실험예에 대해서만 한정되는 것은 아니고, 이와 등가의 기술적 사상의 변형까지를 모두 포함한다.Hereinafter, the contents of the present invention will be described in more detail through the following examples and experimental examples. However, the scope of the present invention is not limited to the following examples and experimental examples, and includes all modifications of the technical idea equivalent thereto.
[실시예 1 : 미생물로부터 생산된 다당체인 알파글루칸 생산][Example 1: Production of alpha-glucan, a polysaccharide produced from microorganisms]
본 실시예에서는 그람양성, 무포자 형성균인 웨이셀라 콘푸사 (Weissella confusa) DSM 20196 및 웨이셀라 사이베리아 (Weissella cibaria) DSM 14295를 활용하여 알파글루칸을 생산하고자 하였으며, 그 방법은 아래와 같았다.In this example, alpha-glucan was attempted to be produced using Gram-positive, spore-forming bacteria Weissella confusa DSM 20196 and Weissella cibaria DSM 14295, and the method was as follows.
미생물체의 성장을 촉진시키기 위해 최적의 성장 조건을 가진 배지(4 g yeast extract, 2 g peptone, 0.2 g MgSO4·7H2O, 0.01 g FeSO4·7H2O, 0.01 g NaCl, 0.01 g MnSO4·H2O, 0.015 g CaCl2·2H2O, and 2 g K2HPO4)를 1 L의 증류수에 섞은 후 탄소원으로써 필요한 자당과 맥아당을 배지 내 1% 첨가한 후 상기 웨이셀라(Weissella) 미생물을 각각 접종하고, 37℃에서 16시간 배양하였다. 웨이셀라(Weissella) 미생물 배양액은 원심분리기를 이용하여 8,000 × g 에 15분 원심분리하여 죽은 균을 제거해준 후, 수용성인 다당체를 얻기 위해 2배 부피의 에탄올(99.9%)을 수용성 다당체가 녹아있는 배양액에 첨가하여 24시간 4℃에서 침전시켰다. 수득한 다당체(EPS)는 투석막(dialysis membrane)(MWCO, Molecular Weight Cut Off: 3,500)을 이용한 투석처리를 통해 단백질, 배지, 올리고당 등을 제거한 후, 동결건조 하였다.To promote the growth of microorganisms, a medium with optimal growth conditions (4 g yeast extract, 2 g peptone, 0.2 g MgSO 4 ·7H 2 O, 0.01 g FeSO 4 ·7H 2 O, 0.01 g NaCl, 0.01 g MnSO 4 ·H 2 O, 0.015 g CaCl 2 ·2H 2 O, and 2 g K 2 HPO 4 ) were mixed in 1 L of distilled water, 1% of sucrose and maltose required as carbon sources were added to the medium, and then the Weissella Each microorganism was inoculated and cultured at 37°C for 16 hours. Weissella microbial culture medium was centrifuged at 8,000 It was added to the culture medium and allowed to precipitate at 4°C for 24 hours. The obtained polysaccharide (EPS) was subjected to dialysis using a dialysis membrane (MWCO, Molecular Weight Cut Off: 3,500) to remove proteins, media, oligosaccharides, etc., and then freeze-dried.
[실험예 1 : 미생물로부터 생산된 알파글루칸 구조 특성 분석][Experimental Example 1: Analysis of structural characteristics of alpha-glucan produced from microorganisms]
본 실험에서는 상기 실시예 1에서 제조한 웨이셀라 콘푸사 (Weissella confusa) DSM 20196 또는 웨이셀라 사이베리아 (Weissella cibaria) DSM 14295로부터 생산된 다당체의 구조 특성을 분석하고자 하였다.In this experiment, we attempted to analyze the structural characteristics of polysaccharides produced from Weissella confusa DSM 20196 or Weissella cibaria DSM 14295 prepared in Example 1.
(1) 고성능 음이온 교환 크로마토그래피 (High performance anion exchange chromatography, HPAEC)을 이용한 구성당 분석 (1) Constituent sugar analysis using high performance anion exchange chromatography (HPAEC)
도 1은 웨이셀라 콘푸사 (Weissella confusa) DSM 20196 및 웨이셀라 사이베리아 (Weissella cibaria) DSM 14295를 이용하여 미생물로부터 생성되는 다당체의 구성당을 확인한 결과를 나타낸 것으로, 도 1에서 보듯이, 상기 두 미생물로부터 생성된 두 가지의 다당체 모두 포도당을 가리키는 단일 피크를 확인할 수 있었으며, 이를 통해 이 두개의 다당체는 글루칸인 것으로 판단되었다.Figure 1 shows the results of confirming the constituent sugars of polysaccharides produced from microorganisms using Weissella confusa DSM 20196 and Weissella cibaria DSM 14295. As shown in Figure 1, the two A single peak indicating glucose was identified in both polysaccharides produced from microorganisms, and through this, it was determined that these two polysaccharides were glucans.
(2) 고성능 크기배제 크로마토그래피(High performance size exclusion chromatography, HPSEC)와 레이저 광 산란 검출기 (Multi-angle laser-light scattering, MALS)를 이용한 분자량 분석(2) Molecular weight analysis using high performance size exclusion chromatography (HPSEC) and multi-angle laser-light scattering (MALS)
아래 표 1은 생산된 웨이셀라(Weissella) 유래 글루칸 타입 다당체의 분자량을 HPSEC-MALS를 통해 확인한 결과를 나타낸 것으로, 웨이셀라 콘푸사 (Weissella confusa) DSM 20196과 웨이셀라 사이베리아 (Weissella cibaria) DSM 14295는 각각 1.03ⅹ108 와 6.47ⅹ107 g/mol의 분자량을 나타냈다. 즉, 웨이셀라 콘푸사 (W.confusa)가 웨이셀라 사이베리아 (W.cibaria)보다 생산된 다당체의 분자량이 더 큰 것으로 나타났다. Table 1 below shows the results of confirming the molecular weight of the produced Weissella -derived glucan-type polysaccharide through HPSEC-MALS, Weissella confusa DSM 20196 and Weissella cibaria DSM 14295. showed molecular weights of 1.03ⅹ10 8 and 6.47ⅹ10 7 g/mol, respectively. In other words, the molecular weight of the polysaccharide produced from Weissella confusa ( W.confusa ) was found to be larger than that of Weissella cybaria ( W.cibaria ).
(3) 푸리에 변환 적외선 분광법 (Fourier Transform Infrared Spectroscopy, FT-IR) 분석(3) Fourier Transform Infrared Spectroscopy (FT-IR) analysis
도 2는 웨이셀라 콘푸사 (W.confusa)와 웨이셀라 사이베리아 (W.cibaria)로부터 생산된 글루칸 타입 다당체를 푸리에 변환 적외선 분광법(FT-IR)을 이용하여 확인한 결과를 나타낸 것이다. 도 2에서 보듯이, 상업용 덱스트란과 비교하여 2개의 다당체 모두 비슷한 주파수에서 공명을 보였으며 다당류의 구조에 수산기 그룹을 갖고 알파-피라노오스(α-pyranose)의 형태를 가진 것으로 확인되었다. 또한, 스펙트럼을 통해 α-1,6 결합과 α-1,3 결합은 감지되었으나 β-결합은 감지되지 않아 결과적으로 알파글루칸의 형태를 확인할 수 있었다.Figure 2 shows the results of confirming glucan-type polysaccharides produced from Weissella confusa ( W.confusa ) and Weissella cyberia ( W.cibaria ) using Fourier transform infrared spectroscopy (FT-IR). As shown in Figure 2, compared to commercial dextran, both polysaccharides showed resonance at similar frequencies and were confirmed to have a hydroxyl group in the polysaccharide structure and to be in the form of alpha-pyranose. In addition, α-1,6 bond and α-1,3 bond were detected through the spectrum, but β-bond was not detected, resulting in confirmation of the form of alpha-glucan.
(4) 핵자기공명기법 (Nuclear magnetic resonance, NMR) 분석(4) Nuclear magnetic resonance (NMR) analysis
웨이셀라(Weissella) 종에서 다당체의 결합구조는 1H NMR 과 13C NMR 분광법이 결합된 2D NMR- HSQC(Heteronuclear Single Quantum Coherence)를 통해 분석되었다. 도 3은 생산된 2개의 다당체를 HSQC를 이용하여 구조를 분석한 결과를 나타낸 것으로, 도 3에서 보듯이 웨이셀라 콘푸사 (W.confusa) 및 웨이셀라 사이베리아 (W.cibaria)의 피크는 상업용 덱스트란과 동일한 모양을 나타냈으며, 탄소와 수소의 결합을 통해 α-1,6의 결합이 우세한 점을 확인할 수 있었다.The binding structure of the polysaccharide in Weissella species was analyzed through 2D NMR-HSQC (Heteronuclear Single Quantum Coherence) combining 1 H NMR and 13 C NMR spectroscopy. Figure 3 shows the results of structural analysis of the two produced polysaccharides using HSQC. As shown in Figure 3, the peaks of Weissella confusa ( W.confusa ) and Weissella cyberia ( W.cibaria ) are commercially It had the same shape as dextran, and it was confirmed that the α-1,6 bond was dominant through the bond between carbon and hydrogen.
즉, 상기 실험 (3)의 푸리에 변환 적외선 분광법과 핵자기공명기법을 이용한 실험 결과, 웨이셀라 콘푸사 (Weissella confusa) DSM 20196 과 웨이셀라 사이베리아 (Weissella cibaria) DSM 14295로부터 생산된 다당체는 α-1,6 결합을 가지며 알파-피라노오스(α-pyranose) 형태의 포도당으로 이루어진 알파글루칸임을 확인할 수 있었다.That is, as a result of the experiment using Fourier transform infrared spectroscopy and nuclear magnetic resonance technique in experiment (3), the polysaccharides produced from Weissella confusa DSM 20196 and Weissella cibaria DSM 14295 were α- It was confirmed that it was alpha-glucan, which has a 1,6 bond and is made of glucose in the form of alpha-pyranose.
(5) 기체 크로마토그래피 질량분석법(Gas Chromatography Mass Specrometry, GC-MS)(5) Gas Chromatography Mass Spectrometry (GC-MS)
기체 크로마토그래피 질량분석법을 이용하여 웨이셀라 콘푸사 (Weissella confusa) DSM 20196 과 웨이셀라 사이베리아 (Weissella cibaria) DSM 14295로부터 생산된 다당체의 결합 비율을 확인하였다 (표 2). 분석 결과, 아래 표 2에서 보듯이 α-1,6 결합이 우세한 형태로 나타났으며, α-1,3과 α-1,4는 각각 약 3%, 1%로 분지되어 있는 형태로 나타났다. 한편, 웨이셀라 콘푸사 (W.confusa)와 웨이셀라 사이베리아 (W.cibaria) 사이에 결합에 대한 유의한 차이는 보이지 않았다. 한편, 글루칸의 유형으로는 덱스트란(dextran, >50% α-1,6), 뮤탄(mutan, 50% α-1,6), 알터난(alternan, α-1,3과 α-1,6의 교차형태)과 루테란(reuteran, 대부분 α-1,4와 약간의 α-1,6)이 있는데 웨이셀라 콘푸사 (W.confusa)와 웨이셀라 사이베리아 (W.cibaria)로부터 생산된 다당체는 그 중에서 덱스트란(dextran)의 형태를 가진 알파글루칸으로 확인되었다.The binding ratio of polysaccharides produced from Weissella confusa DSM 20196 and Weissella cibaria DSM 14295 was confirmed using gas chromatography mass spectrometry (Table 2). As a result of the analysis, as shown in Table 2 below, the α-1,6 bond appeared to be the dominant form, and α-1,3 and α-1,4 appeared to be branched at about 3% and 1%, respectively. Meanwhile, there was no significant difference in binding between Weissella confusa ( W.confusa ) and Weissella cyberia ( W.cibaria ). Meanwhile, the types of glucans include dextran (>50% α-1,6), mutan (50% α-1,6), and alternan (α-1,3 and α-1, 6) and reuteran (mostly α-1,4 and some α-1,6), produced from W.confusa and W.cibaria . Among the polysaccharides, alpha-glucan in the form of dextran was identified.
[실험예 2 : 포도당 생성 속도 (분해 속도) 확인][Experimental Example 2: Confirmation of glucose production rate (decomposition rate)]
본 실험에서는 상기 실시예 1에서 제조한 웨이셀라 콘푸사 (Weissella confusa) DSM 20196 및 웨이셀라 사이베리아 (Weissella cibaria) DSM 14295로부터 생산된 다당체의 알파-글루코시다아제(α-glucosidase)에 의한 포도당 생성 속도 (분해 속도)를 확인하고자 하였다. In this experiment, glucose production by alpha-glucosidase of polysaccharides produced from Weissella confusa DSM 20196 and Weissella cibaria DSM 14295 prepared in Example 1. We wanted to check the speed (decomposition rate).
도 4는 웨이셀라 콘푸사 (Weissella confusa) DSM 20196 과 웨이셀라 사이베리아 (Weissella cibaria) DSM 14295로부터 생산된 다당체를 식이섬유 정량에 사용되는 곰팡이유래 소화효소인 글루코아밀라아제(glucoamylase)와 포유류 유래 소화효소인 알파-글루코시다아제(α-glucosidase)를 이용하여 소화속도를 확인한 결과를 나타낸 것이다. Figure 4 shows polysaccharides produced from Weissella confusa DSM 20196 and Weissella cibaria DSM 14295 using glucoamylase, a fungal-derived digestive enzyme used to quantify dietary fiber, and mammalian-derived digestive enzymes. This shows the results of checking the digestion speed using α-glucosidase.
도 4에서 보듯이, 곰팡이 유래 소화효소인 글루코아밀라아제(glucoamylase)에서는 본 발명 웨이셀라 (Weissella) 미생물 유래 알파글루칸들이 분해되지 않은 반면, 포유류 유래 소화효소 수준에서는 대조군인 말토덱스트란(Maltodextran) DE1에 비해 더 천천히 소화되는 것을 확인할 수 있었다. As shown in Figure 4, alpha-glucans derived from the Weissella microorganism of the present invention were not decomposed by glucoamylase, a digestive enzyme derived from fungi, whereas at the level of digestive enzymes derived from mammals, they were decomposed in maltodextran DE1, a control group. It was confirmed that it was digested more slowly.
웨이셀라 콘푸사 (W.confusa) 유래 알파글루칸과 웨이셀라 사이베리아 (W.cibaria) 유래 알파글루칸은 12시간까지 유사하게 가수분해되다가 24시간 이후부터는 웨이셀라 콘푸사 (W.confusa) 유래 알파글루칸보다 웨이셀라 사이베리아 (W.cibaria) 유래 알파글루칸이 더 가수분해율이 높았다. 이는 웨이셀라 콘푸사 (W.confusa)로부터 생산된 다당체가 웨이셀라 사이베리아 (W.cibaria)보다 분자량이 높기 때문이라고 판단할 수 있었다. Alpha-glucan derived from Weissella confusa ( W.confusa ) and alpha-glucan derived from Weissella cibaria ( W.cibaria ) are hydrolyzed similarly for up to 12 hours, but after 24 hours, alpha-glucan derived from Weissella confusa ( W.confusa ) Alpha-glucan derived from Weissella ciberia ( W.cibaria ) had a higher hydrolysis rate. It could be determined that this was because the polysaccharide produced from W.confusa had a higher molecular weight than that of W.cibaria .
한편, 실험에 같이 사용된 상업용 덱스트란은 분자량이 낮아 (Mw; 64,000 Da) 효소에 공격을 받기 쉽기 때문에 더 빨리 가수분해 되는 것으로 판단되었다. 이러한 결과는 본 발명 웨이셀라 콘푸사 (W.confusa) 및 웨이셀라 사이베리아 (W.cibaria)와 같은 웨이셀라(Weissella) 미생물로부터 생산된 알파글루칸이, 소화되지 않는 난소화성 식이섬유의 소재가 아닌 천천히 분해되어 지속적으로 에너지를 공급할 수 있는 지소화성 소재임을 의미하는 것이다. Meanwhile, the commercial dextran used in the experiment was judged to be hydrolyzed more quickly because it had a low molecular weight (Mw; 64,000 Da) and was easily attacked by enzymes. These results show that alpha-glucan produced from Weissella microorganisms such as W.confusa and W.cibaria of the present invention is not a material of indigestible dietary fiber. This means that it is a low-digestion material that decomposes slowly and can continuously supply energy.
즉, 본 연구를 통해 웨이셀라 콘푸사 (W.confusa)와 웨이셀라 사이베리아 (W.cibaria)로부터 생산된 다당체는 α-1,6 branch linkage가 많이 존재하는 웨이셀라(Weissella) 미생물 유래 알파글루칸으로써, 이러한 물질은 분자량이 높고 포도당으로의 가수분해 속도를 늦출 수 있어 지소화성 기능성 물질로 응용될 수 있을 것으로 판단되었다. 이와 같이, 웨이셀라(Weissella) 종의 다당체는 천천히 소화되는 성질을 가진 기능성 원료로 식품산업에서 응용 가능성이 있음을 확인하였다.In other words, through this study, polysaccharides produced from Weissella confusa ( W.confusa ) and Weissella cyberia ( W.cibaria ) are alpha-glucans derived from Weissella microorganisms, which contain a lot of α-1,6 branch linkage. As a result, it was judged that this material could be applied as a lipolytic functional material because it has a high molecular weight and can slow down the rate of hydrolysis to glucose. In this way, it was confirmed that the polysaccharide of Weissella species has the potential for application in the food industry as a functional raw material with slowly digestible properties.
[제조예 1 : 지소화성 알파글루칸 식품 산업 적용][Preparation Example 1: Application of digestible alpha-glucan to food industry]
본 제조예에서는 웨이셀라 콘푸사 (Weissella confusa) DSM 20196 과 웨이셀라 사이베리아 (Weissella cibaria) DSM 14295로부터 생산된 지소화성 알파글루칸을 죽에 첨가하여 식품 산업에 적용하고자 하였다.In this production example, the digestible alpha-glucan produced from Weissella confusa DSM 20196 and Weissella cibaria DSM 14295 was added to porridge and applied to the food industry.
아래 표 3은 웨이셀라 콘푸사 (Weissella confusa) DSM 20196 과 웨이셀라 사이베리아 (Weissella cibaria) DSM 14295로부터 생산된 지소화성 알파글루칸을 기능성 원료로 적용하기 위해 죽에 알파글루칸을 첨가하여 점도 변화를 측정한 결과이다. Table 3 below shows the viscosity change measured by adding alpha-glucan to porridge in order to apply the digestible alpha-glucan produced from Weissella confusa DSM 20196 and Weissella cibaria DSM 14295 as a functional raw material. This is one result.
표 3에서 보듯이, 상업용 덱스트란과 웨이셀라 콘푸사 (W.confusa)는 쌀의 함량이 줄어들고 다당체(EPS) 함량이 늘어남에 따라 점도가 낮아지는 것으로 나타났으나, 웨이셀라 사이베리아 (W.cibaria)의 경우에는 쌀의 함량이 줄어들고 다당체의 함량이 늘어남에 따라 점도가 일정하게 증가하는 것으로 나타났다. As shown in Table 3, the viscosity of commercial dextran and Weissella confusa ( W. confusa ) was found to decrease as the rice content decreased and the polysaccharide (EPS) content increased, but Weissella siberia ( W. confusa ) showed a decrease in viscosity. cibaria ), the viscosity was found to increase consistently as the rice content decreased and the polysaccharide content increased.
한편, 생산된 죽을 24시간 실온에 방치한 후의 점도를 측정하여 노화도를 측정한 결과, 대부분의 글루칸을 쌀 대신에 첨가하였을 때 점도가 쌀만 들어있는 죽에 비해 점도가 낮아 노화가 일어나지 않은 점을 확인할 수 있었다. 즉, 쌀 대신에 다당체(EPS)를 첨가하여도 점도가 일정하게 유지되면서 지소화성의 특징에 따라 생체 내에 지속적인 에너지 전달 및 인슐린의 급격한 농도 상승 억제를 통한 대사 과정의 항상성을 확보할 수 있었다.Meanwhile, as a result of measuring the degree of aging by measuring the viscosity of the produced porridge after leaving it at room temperature for 24 hours, it was confirmed that when most glucans were added instead of rice, the viscosity was lower than that of porridge containing only rice, so aging did not occur. I was able to. In other words, even when polysaccharide (EPS) was added instead of rice, the viscosity remained constant, and homeostasis of the metabolic process was secured through continuous energy transfer in the body and suppression of rapid rise in insulin concentration according to the characteristics of lipolysis.
Claims (4)
상기 단계 (a)에서 제조한 배양액에 웨이셀라 콘푸사 (Weissella confusa) 및 웨이셀라 사이베리아 (Weissella cibaria) 중 선택되는 어느 하나의 웨이셀라(Weissella) 속 균주를 접종하는 단계 (b);
상기 단계 (b)에서 웨이셀라(Weissella) 속 균주를 접종한 배양액을 원심분리하여 사균을 제거하는 단계 (c);
상기 단계 (c) 후, 배양액에 에탄올을 첨가하여 다당체를 침전시키는 단계 (d);
상기 단계 (d) 후, 상기 다당체가 침전된 배양액을 투석처리하여 다당체를 수득하는 단계 (e);를 포함하는 지소화성 알파글루칸의 제조방법.
Step (a) of preparing a culture medium by mixing the medium and distilled water and then adding sucrose and maltose;
(b) inoculating the culture medium prepared in step (a) with a strain of the Weissella genus selected from Weissella confusa and Weissella cibaria ;
(c) removing dead cells by centrifuging the culture medium inoculated with the Weissella genus strain in step (b);
After step (c), adding ethanol to the culture medium to precipitate the polysaccharide (d);
After step (d), a step (e) of obtaining a polysaccharide by dialyzing the culture medium in which the polysaccharide has been precipitated.
상기 투석처리는,
MWCO (Molecular Weight Cut Off) 값이 3,500인 투석막(dialysis membrane)을 이용하여 수행하는 것을 특징으로 하는 지소화성 알파글루칸의 제조방법.
According to paragraph 1,
The dialysis treatment,
A method for producing lipotropic alpha-glucan, characterized in that it is performed using a dialysis membrane with a MWCO (Molecular Weight Cut Off) value of 3,500.
A food composition comprising lipodigestible alpha-glucan prepared by the method of claim 1.
상기 식품은,
당뇨병 환자용 식이인 것을 특징으로 하는 식품 조성물. According to paragraph 3,
The above foods are:
A food composition characterized in that it is a diet for diabetic patients.
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| KR101764287B1 (en) | 2015-11-19 | 2017-08-14 | 안동대학교 산학협력단 | Non-digestible dehydrated potato parenchyma cells by enzymatic alpha-glucan coating and manufacturing method thereof |
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