KR20210098570A - Manufacturing method of isomalto-oligosaccharide using rice powder - Google Patents
Manufacturing method of isomalto-oligosaccharide using rice powder Download PDFInfo
- Publication number
- KR20210098570A KR20210098570A KR1020200011731A KR20200011731A KR20210098570A KR 20210098570 A KR20210098570 A KR 20210098570A KR 1020200011731 A KR1020200011731 A KR 1020200011731A KR 20200011731 A KR20200011731 A KR 20200011731A KR 20210098570 A KR20210098570 A KR 20210098570A
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- KR
- South Korea
- Prior art keywords
- isomaltooligosaccharide
- rice flour
- rice
- bondage
- temperature
- Prior art date
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Abstract
Description
본 발명은 쌀가루를 이용한 이소말토올리고당의 제조방법에 관한 것이다. The present invention relates to a method for producing isomaltooligosaccharide using rice flour.
인간의 장관 내에는 400여종 이상의 세균이 대장 내용물 g당 100조 정도의 숫자로 서식하고 있으며 일정한 균형을 유지하는 복잡한 생태계로 구성되어 있다. 장내 균총은 유익균과 유해균으로 나눌 수 있으며, 이들의 균형에 의해 건강상태가 조절된다. 건강을 유지하기 위해서는 유익균이 많고 유해균이 적은 상태로 장내 균총을 유지시켜야 한다. 균총은 외부에서 유입된 음식물이나 스트레스, 호르몬 분비 등 인체의 상태에 따라 변화될 수 있으며, 이들이 생성하는 효소와 대사산물들은 여러 가지 물질의 인체 내 대사에 관여하고, 영양분이나 약물의 흡수, 독성물질의 생성에 영향을 미친다.In the human intestinal tract, more than 400 kinds of bacteria inhabit in the number of about 100 trillion per gram of the contents of the large intestine, and it is composed of a complex ecosystem that maintains a certain balance. The intestinal flora can be divided into beneficial bacteria and harmful bacteria, and the health status is controlled by the balance between them. In order to maintain health, it is necessary to maintain the intestinal flora in a state where there are many beneficial bacteria and few harmful bacteria. The flora can change depending on the state of the human body, such as food, stress, hormone secretion, etc. introduced from the outside affects the creation of
이러한 장내 유익균을 프로바이오틱스라 하는데 프리바이오틱스는 프로바이오틱스를 활성화하는 동시에 장내 유해균을 억제하는 성분으로, 프로바이오틱스가 잘 자랄 수 있도록 장내 환경을 조성하는 역할을 한다. 즉, 프리바이오틱스는 우리 몸에 유익한 역할을 하는 미생물인 프로바이오틱스의 먹이 역할을 한다.These beneficial bacteria in the gut are called probiotics. Prebiotics are ingredients that activate probiotics and inhibit harmful bacteria in the gut, and play a role in creating an intestinal environment for probiotics to thrive. In other words, prebiotics serve as food for probiotics, which are microorganisms that play a beneficial role in our body.
장내 유익균 균총의 증가는 여러 가지 질병 중에서도 설사나 복부팽만 등을 동반하는 과민성 대장 증후군에 직접적인 영향을 끼친다. 과민성 대장 증후군은 다른 질환이나 해부학적 이상 없이 대장 근육의 과민해진 수축 운동으로 인한 기능 장애로 발생하는 증상들의 총칭이며, 과민성 대장 증후군의 명확한 원인은 아직 밝혀진 것이 없지만, 현대인에게 있어서는 과민성 대장 증후군의 발생이 주로 스트레스로 인해 유발된다는 다양한 연구결과들이 보고되고 있다. The increase in beneficial intestinal flora has a direct effect on irritable bowel syndrome accompanied by diarrhea or abdominal distension among various diseases. Irritable Bowel Syndrome is a generic term for symptoms caused by dysfunction caused by overactive contraction of the colon muscle without other diseases or anatomical abnormalities. Various studies have reported that this is mainly caused by stress.
쌀은 인류가 석기를 사용하던 때부터 지금까지'에너지의 원천'이자 '문화의 근간'으로 기능해왔으며, 밀, 보리와 함께 세계 3대 곡물의 하나로서, 현재 전 세계 약 30억의 인구가 쌀을 주식으로 하고 있으나, 쌀 재배는 노동 집약적 특성과 영양학적 완전성을 지니고 있다고 알려져 있다. 세계 경지 면적의 약 20%가 논으로 쌀은 타 작물에 비해 인구 부양력이 커서 쌀을 주식으로 하는 지역은 인구밀도가 높은 편에 속하지만, 밀의 경지 면적은 전체 32%로 세계 1위이나 쌀보다 인구 부양력이 낮아 전 세계 인구의 10%만이 주식으로 이용하고 있다. Rice has been functioning as a 'source of energy' and 'a foundation of culture' since the time humans used stone tools. It is one of the world's three major grains along with wheat and barley. Although rice is the staple food, rice cultivation is known to have labor-intensive characteristics and nutritional integrity. About 20% of the world's arable land is paddy field, and rice has a higher population buoyancy compared to other crops, so the area where rice is the mainstay has a high population density. Due to its low population support capacity, only 10% of the world's population uses it as a stock.
쌀은 한국인의 주요 에너지원으로써 성인이 하루에 필요한 에너지의 30~40%를 쌀에서 섭취한다. 쌀에는 탄수화물, 단백질, 지방, 식이섬유, 미네랄 등 10여 가지 영양성분이 존재하며 쌀눈과 쌀겨에 주로 함유되어 있으며, 최근 쌀에 함유된 고유의 기능에 대한 관심이 높아지면서 다양한 기능성 쌀의 연구개발이 활발하게 진행되고 있고, 전통육종, 생명공학 기술, 발아 처리를 통해 쌀의 기능성 성분 함량을 증가시키는 품종 개발과 가공 기술 발전이 가속화된다. Rice is the main energy source for Koreans, and 30-40% of the daily energy required by adults is from rice. Rice contains about 10 nutritional components such as carbohydrates, proteins, fats, dietary fiber, and minerals, and is mainly contained in rice bran and rice bran. This is actively progressing, and the development of varieties and processing technologies that increase the content of functional ingredients in rice through traditional breeding, biotechnology, and germination treatment are accelerating.
그러나, 식생활의 다양화 및 식습관 변화로 인하여 쌀 소비량은 매년1.6~1.2kg씩 감소하고 있는 추세이며, 통계청의 양곡소비량 통계조사 결과 2002년인당 87kg 소비량 대비 2013년 인당 68.5kg으로 약 21.3% 감소한 것으로 나타났으며 2014년에는 인당 65.1kg 소비량까지 감소한 것으로 나타났다. However, due to the diversification of dietary life and changes in eating habits, rice consumption is on the decline by 1.6~1.2kg per year. In 2014, consumption per person decreased to 65.1 kg.
게다가 MMA(의무수입물량)를 통해 들어오는 쌀로 인하여 국내산 쌀의 잉여량이 증가하고 있는 상황인데, 쌀 재고량 추이를 살펴보면, 2010년 쌀 재고량은 143만 4000톤으로 가장 높은 수치를 보이다가 이후 2012년까지 재고량이 다소 감소하였지만 2012년 이후 지속적으로 쌀 재고량이 늘어나 2015년 9월 쌀 재고량은 2012년 대비 95% 정도가 상승하여 136만톤으로 나타났고, 쌀 재고의 증가로 쌀 재고 10만 톤당 연간 소요되는 비용이 3,131억(2012년)에서 4,297억(2015년 9월)이 되어 경제적 손실이 계속 증가하고 있다. 그러나 가공식품 제조에 이용되는 쌀은 국내 생산량의 약 6%에 불과해 식량으로서의 쌀이 아닌 다른 용도로의 쌀가공 산업의 필요성이 강하게 요구되는 실정이다. 따라서, 최근의 식품 소비 패턴인 간편식, Well-Being, LOHAS, 환경과 함께 쌀 가공제품 소비 증가와 같은 새로운 소비패턴이 형성될 필요가 있다. In addition, the surplus of domestic rice is increasing due to rice coming in through MMA (mandatory import quantity). Although this has decreased slightly, the rice inventory has continuously increased since 2012, and the rice inventory in September 2015 increased by 95% compared to 2012 to 1.36 million tons. From 313.1 billion (2012) to 429.7 billion (September 2015), the economic loss continues to increase. However, rice used for manufacturing processed foods only accounts for about 6% of domestic production, so there is a strong need for the rice processing industry for purposes other than rice as food. Therefore, it is necessary to form a new consumption pattern such as increased consumption of processed rice products along with convenience food, well-being, LOHAS, and the environment, which are recent food consumption patterns.
이소말토올리고당은 포도당 분자가 α-1,6 결합을 하고 있는 분지올리고당으로, 당 전이 효소에 의해 맥아당이나 말토올리고당의 전이반응으로 만들어진다. 말토올리고당은 포도당 3~10개가 α-(1→4) 결합으로 연결된 직쇄 올리고당이다. 이소말토올리고당으로는 대표적으로 isomaltose, panose, isomaltotriose, isomaltotetraose 등이 있다. 또한 이소말토올리고당은 열 안정성, 저점도, 높은 보습성 등의 특징으로 식품에 광범위하게 응용이 가능하고 효모에 의해 발효되지 않아 베이킹이나 주정, 조미료 등에 감미료로 사용이 가능하다. 이소말토올리고당의 생리학적 기능성으로는 장내 효소에 의해 서서히 분해되기 때문에 혈당지수를 낮추며, 장내 유익균들을 증식시켜 정장작용을 촉진시키는 프리바이오틱스 기능이 있으며, 면역기능을 향상시키며 일부 미네랄의 생체 이용률을 높이고, 중성지방의 신진대사를 촉진하고, 혈중 콜레스테롤을 개선하는 등의 효과가 있다고 보고되었다. Isomaltooligosaccharide is a branched oligosaccharide in which glucose molecules have α-1,6 bonds, and is produced by transfer reaction of maltose or maltooligosaccharide by a sugar transferase. Maltooligosaccharide is a straight-chain oligosaccharide in which 3 to 10 glucose units are linked by α-(1→4) bonds. Representative examples of isomaltooligosaccharides include isomaltose, panose, isomaltotriose, and isomaltotetraose. In addition, isomaltooligosaccharide can be widely applied to food due to its characteristics such as thermal stability, low viscosity, and high moisture retention, and it is not fermented by yeast, so it can be used as a sweetener for baking, alcohol, and seasoning. The physiological function of isomaltooligosaccharide is that it is slowly decomposed by intestinal enzymes, so it lowers the glycemic index, has a prebiotic function that promotes intestinal function by proliferating beneficial intestinal bacteria, improves immune function, and improves the bioavailability of some minerals. It has been reported to have effects such as increasing blood pressure, promoting the metabolism of triglycerides, and improving blood cholesterol.
한편, 본 발명자들은 쌀 소비량 증대 및 프리바이오틱스를 이용한 과민성 대장 증후군의 개선용 조성물에 대해 연구하던 중, 쌀가루와 각종 효소들의 반응시간 및 반응온도 등을 조절하여 기존보다 프리바이오틱스 기능이 현저하게 강화된 이소말토올리고당의 제조가 가능함을 확인하여 본 발명을 완성하였다. On the other hand, while the present inventors were studying a composition for improving rice consumption and irritable bowel syndrome using prebiotics, the prebiotic function was significantly improved than before by controlling the reaction time and reaction temperature of rice flour and various enzymes. The present invention was completed by confirming that it is possible to prepare a reinforced isomaltooligosaccharide.
본 발명의 목적은 쌀가루를 이용한 이소말토올리고당의 제조방법을 제공하는 데에 있다. It is an object of the present invention to provide a method for producing isomaltooligosaccharide using rice flour.
본 발명은 쌀가루를 이용한 이소말토올리고당의 제조방법에 관한 것이다. The present invention relates to a method for producing isomaltooligosaccharide using rice flour.
상기 방법은 바람직하게는 하기와 같다. The method is preferably as follows.
즉, (제1단계) 쌀가루 분산액에 칼슘이온이 60~80ppm이 되도록 CaCO3를 첨가하고 0.8~1.2N의 HCl을 점적하면서 pH를 6.0~6.4로 조정하여 쌀가루 슬러리를 제조하는 단계; That is, (first step) adding CaCO 3 so that calcium ions are 60 to 80 ppm to the rice flour dispersion, and adjusting the pH to 6.0 to 6.4 while adding 0.8 to 1.2 N of HCl dropwise to prepare a rice flour slurry;
(제2단계) 상기 쌀가루 슬러리를 승온시켜 58~62℃에서 α-아밀라아제를 첨가하고, 상기 효소 첨가 후 쌀가루 슬러리의 온도를 계속 승온시켜 90~95℃로 온도를 유지한 상태에서 효소 반응하여 쌀가루 슬러리를 액화하는 단계;(Step 2) The temperature of the rice flour slurry is raised and α-amylase is added at 58 to 62 ° C. After the enzyme is added, the temperature of the rice flour slurry is continuously raised to maintain the temperature at 90 to 95 ° C. liquefying the slurry;
(제3단계) 제2단계에서 얻은 쌀가루 액화액의 온도를 55~60℃로 하강하여 상기 온도를 유지한 상태에서, β-아밀레아제, 풀루라나아제 및 트랜스글루코시다아제를 쌀 액화액에 첨가하고 반응하여 상기 액화액을 당화함으로써 이소말토올리고당을 얻는 단계; (3rd step) β-amylase, pullulanase and transglucosidase were added to the rice liquefied solution while maintaining the temperature by lowering the temperature of the rice flour liquefied liquid obtained in the second step to 55-60 ° C. adding and reacting to saccharify the liquid to obtain isomaltooligosaccharide;
(제4단계) 제3단계에서 얻은 이소말토올리고당을 여과 및 농축하여 70~80 브릭스 당도의 이소말토올리고당을 얻는 단계;일 수 있다. (Step 4) Filtering and concentrating the isomaltooligosaccharide obtained in the third step to obtain isomaltooligosaccharide having a sugar content of 70 to 80 Brix; may be.
상기 제1단계에서 쌀가루 분산액은 쌀 100 중량부 기준 물 300~500 중량부가 혼합되어 분산된 것을 특징으로 한다. In the first step, the rice flour dispersion is characterized in that 300 to 500 parts by weight of water are mixed and dispersed based on 100 parts by weight of rice.
상기 제1단계에서 쌀가루의 입자크기는 180μm 이하이며, 수분함량은 14중량% 이하인 것이 좋다. 바람직하게는 입자크기 10~180μm이며, 수분함량은 12~14중량%일 수 있다. 쌀가루의 입자크기가 180μm를 초과하게 되면 당화반응이 잘 일어나지 않아 바람직하지 않으며, 불린 쌀이나 덜 마른 쌀을 이용하여 수분함량이 더 높은 쌀가루를 사용하여도 쌀가루 분산액을 제조하기 쉽지 않아 당화반응이 잘 일어나지 않을 수 있다. In the first step, the particle size of the rice flour is 180 μm or less, and the moisture content is preferably 14 wt % or less. Preferably, the particle size is 10 to 180 μm, and the moisture content may be 12 to 14 wt %. If the particle size of the rice flour exceeds 180μm, saccharification reaction does not occur well, which is undesirable. may not happen
상기 제2단계에서 사용하는 α-아밀라아제는 90~100℃의 온도에서 효소활성이 최적인 내열성을 갖는 것일 수 있다. 이 효소의 제조는 효소 제조를 위해 최적화된 균주라면 어느 것이나 사용할 수 있으나, 본 발명에서는 바람직하게는 바실러스 리케니포르미스 균주로부터 생산된 것을 사용하는 것이 더 좋다. The α-amylase used in the second step may have heat resistance with optimal enzyme activity at a temperature of 90 to 100°C. For the production of this enzyme, any strain optimized for enzyme production may be used, but in the present invention, it is preferable to use one produced from a Bacillus licheniformis strain.
상기 제2단계에서 α-아밀라아제 효소를 58~62℃에서 첨가하여 쌀가루가 급격하게 호화되는 것을 방지한다. 최적 반응온도인 90~95℃로 승온하여 α-아밀라아제 효소를 첨가하게 되면 그 전에 호화가 급격히 일어나 효소반응이 일어나기 이전에 쌀가루가 뭉쳐져 분산액 내에서 덩어리를 형성하며, 이러한 경우 덩어리 내부에서는 효소반응이 일어나지 않을 수 있어 좋지 않다. In the second step, the α-amylase enzyme is added at 58 to 62° C. to prevent rapid gelatinization of rice flour. If the temperature is raised to 90~95℃, which is the optimum reaction temperature, and α-amylase enzyme is added, gelatinization occurs rapidly before the enzymatic reaction, and the rice flour is agglomerated to form a lump in the dispersion. In this case, the enzyme reaction inside the lump It's not good because it can't happen.
상기 제2단계의 90~95℃에서의 효소 반응 시간은 1~3시간 동안 인 것이 적절하다. It is appropriate that the enzymatic reaction time at 90 to 95° C. of the second step is for 1 to 3 hours.
상기 제2단계의 α-아밀라아제 효소는 쌀가루 100 중량부 기준 0.05~0.1 중량부가 포함될 수 있다. The α-amylase enzyme of the second step may contain 0.05 to 0.1 parts by weight based on 100 parts by weight of rice flour.
상기 제3단계의 효소반응은 55~58℃에서 진행하는 것이 좋다. The enzymatic reaction of the third step is preferably carried out at 55 ~ 58 ℃.
상기 제3단계에서 사용하는 효소인 β-아밀레아제는 55~65℃, 풀루라나아제 55~60℃, 트랜스글루코시다아제는 58~62℃에서 각각 최적 활성을 가지며, 이들 효소도 이 효소의 제조는 효소 제조를 위해 최적화된 균주라면 어느 것이나 사용할 수 있으나, 바람직하게는 바실러스 서브틸리스아밀로리퀘파시엔스 균주로부터 생산된 β-아밀레아제, 바실러스 애시도풀루리티커스 균주로부터 생산된 풀루라나아제 및 아스퍼질러스 니거 균주로부터 생산된 트랜스글루코시다아제를 사용하는 것이 당화에 가장 적절하다. The enzyme used in the third step, β-amylase, has optimal activity at 55 to 65 ° C, pullulanase at 55 to 60 ° C, and transglucosidase at 58 to 62 ° C. For the preparation, any strain optimized for enzyme production may be used, but preferably β-amylase produced from a Bacillus subtilis amyloliquefaciens strain, pullulus produced from a Bacillus acidopuluriticus strain The use of transglucosidase produced from Lanase and Aspergillus niger strains is most suitable for glycosylation.
상기 제3단계의 효소의 반응시간은 30~40시간인 것이 바람직하다. The reaction time of the enzyme in the third step is preferably 30 to 40 hours.
상기 제3단계의 각 효소의 함량은 쌀가루 100 중량부를 기준으로, β-아밀레아제 0.001~0.002 중량부, 풀루라나아제 0.05~0.1 중량부 및 트랜스글루코시다아제 0.07~0.2 중량부일 수 있다. 이 농도로 효소를 처리할 때, glucose의 함량은 감소되고 중합도가 높은 이소말토올리고당의 함량이 증가하는 효과가 있다. 이 때 각 효소의 함량 범위 미만일 경우 당화가 잘 일어나지 않아 이소말토올리고당의 생성이 잘 되지 않으며, 각 효소의 함량 범위를 초과한다하더라도 이소말토올리고당의 생성이 증가되지 않고 오히려 더 감소할 수 있어 바람직하지 않다. The content of each enzyme in the third step may be 0.001 to 0.002 parts by weight of β-amylase, 0.05 to 0.1 parts by weight of pullulanase, and 0.07 to 0.2 parts by weight of transglucosidase based on 100 parts by weight of rice flour. When the enzyme is treated at this concentration, the content of glucose is reduced and the content of isomaltooligosaccharide having a high degree of polymerization is increased. At this time, if the content of each enzyme is less than the content range, saccharification does not occur well, so the production of isomaltooligosaccharide is not good. not.
상기 제3단계에서 얻은 이소말토올리고당을 함유한 조성물은 16~18 브릭스인 것을 특징으로 하며, 이소말토오스(isomaltose) 30~40 g/L, 판노스(panose) 8~15 g/L, 이소말토트리오스(isomaltotriose) 15~25 g/L, 이소말토테트라오스(isomaltotetraose) 5~10 g/L, 이소말토펜타오스(isomaltopentaose) 0.5~2 g/L 이고, 총 이소말토올리고당이 60~95g/L, 물과 기타 미량성분을 제외한 총 당류를 100 중량%라 할 때, 이소말토올리고당의 함량은 50~60 중량%일 수 있다. The composition containing isomaltooligosaccharide obtained in the third step is characterized in that it is 16-18 Brix, isomaltose 30-40 g/L, panose 8-15 g/L, isomal 15-25 g/L of totriose, 5-10 g/L of isomaltotetraose, 0.5-2 g/L of isomaltopentaose, and 60-95 g/L of total isomaltooligosaccharides When the total sugar excluding L, water and other minor components is 100 wt%, the content of isomaltooligosaccharide may be 50 to 60 wt%.
상기 제4단계에서 농축 후 얻은 이소말토올리고당은 이소말토오스(isomaltose) 125~166 g/L, 판노스(panose) 33~63 g/L, 이소말토트리오스(isomaltotriose) 63~105 g/L, 이소말토테트라오스(isomaltotetraose) 21~42 g/L, 이소말토펜타오스(isomaltopentaose) 1~8 g/L 이고, 총 이소말토올리고당이 250~395g/L, 물과 기타 미량성분을 제외한 총 당류를 100 중량%라 할 때, 이소말토올리고당의 함량은 50~60 중량%일 수 있다. The isomaltooligosaccharide obtained after concentration in the fourth step is isomaltose (isomaltose) 125 ~ 166 g / L, panose (panose) 33 ~ 63 g / L, isomaltotriose (isomaltotriose) 63 ~ 105 g / L, Isomaltotetraose 21-42 g/L, isomaltopentaose 1-8 g/L, total isomaltooligosaccharides 250-395 g/L, total sugars excluding water and other minor ingredients When referring to 100% by weight, the content of isomaltooligosaccharide may be 50 to 60% by weight.
상기 제4단계에서 농축 후 얻은 이소말토올리고당은 70~80 브릭스일 수 있다.The isomaltooligosaccharide obtained after concentration in the fourth step may be 70 to 80 Brix.
상기 제4단계 이후, (제5단계) 농축된 이소말토올리고당을 분말화하여 분말을 얻는 단계;가 더 포함될 수 있다. After the fourth step, (fifth step) pulverizing the concentrated isomaltooligosaccharide to obtain a powder; may be further included.
본 발명은 상기 방법으로 제조된 이소말토올리고당이 포함된 프리바이오틱스를 제공할 수 있다. The present invention may provide a prebiotic containing isomaltooligosaccharide prepared by the above method.
또한 상기 이소말토올리고당을 함유하는 과민성 대장 증후군의 예방, 치료, 개선용 약학 조성물 또는 건강기능식품으로도 이용가능하다. It is also available as a pharmaceutical composition or health functional food for the prevention, treatment, and improvement of irritable bowel syndrome containing the isomaltooligosaccharide.
본 발명에서 과민성 대장 증후군은 설사 우세형 질환일 수 있다. In the present invention, irritable bowel syndrome may be a diarrhea-predominant disease.
과민성 대장 증후군의 예방 또는 치료용 약학 조성물에 있어서, 프리바이오틱스인 것을 감안하여 이를 경구 투여용 조성물로서 이소말토올리고당을 포함한 그 자체로 이용하는 것이 가장 좋으며, 필요에 따라 각종 부형제를 첨가할 수는 있다. 본 발명의 약학 조성물의 투여량은 치료받을 대상의 연령, 성별, 체중과, 치료할 특정 질환 또는 병리 상태, 질환 또는 병리 상태의 심각도, 투여경로 및 처방자의 판단에 따라 달라질 것이다. 이러한 인자에 기초한 투여량 결정은 당업자의 수준 내에 있으며, 일반적으로 투여량은 0.01㎎/㎏/일 내지 대략 2000㎎/㎏/일의 범위이다. 더 바람직한 투여량은 1㎎/㎏/일 내지 500㎎/㎏/일이다. 투여는 하루에 한번 투여할 수도 있고, 수회 나누어 투여할 수도 있다. 상기 투여량은 어떠한 면으로든 본 발명의 범위를 한정하는 것은 아니다. 본 발명의 약학 조성물은 쥐, 가축, 인간 등의 포유동물에 다양한 경로로 투여될 수 있다. In the pharmaceutical composition for the prevention or treatment of irritable bowel syndrome, it is best to use it by itself, including isomaltooligosaccharide, as a composition for oral administration, considering that it is a prebiotic, and various excipients can be added as needed. . The dosage of the pharmaceutical composition of the present invention will vary depending on the age, sex, and weight of the subject to be treated, the specific disease or pathological condition to be treated, the severity of the disease or pathological condition, the route of administration, and the judgment of the prescriber. Dosage determination based on these factors is within the level of the skilled artisan, and dosages generally range from 0.01 mg/kg/day to approximately 2000 mg/kg/day. A more preferred dosage is 1 mg/kg/day to 500 mg/kg/day. Administration may be administered once a day, or may be administered in several divided doses. The above dosage does not limit the scope of the present invention in any way. The pharmaceutical composition of the present invention may be administered to mammals such as mice, livestock, and humans by various routes.
과민성 대장 증후군의 예방 또는 개선용 건강기능식품은 각종 드링크제, 육류, 소세지, 빵, 캔디류, 스넥류, 면류, 아이스크림, 유제품, 스프, 이온음료, 음료수, 알코올 음료, 껌, 차, 비타민 복합제 등의 식품용 첨가제로도 이용가능하다. Health functional foods for the prevention or improvement of irritable bowel syndrome include various drinks, meat, sausages, bread, candy, snacks, noodles, ice cream, dairy products, soups, ionic drinks, beverages, alcoholic beverages, gum, tea, vitamin complexes, etc. It can also be used as an additive for
본 발명은 쌀가루를 이용한 이소말토올리고당의 제조방법에 관한 것으로서, 쌀가루와 각종 효소들의 반응시간 및 반응온도 등을 조절하여 기존보다 프리바이오틱스 기능이 현저하게 강화된 이소말토올리고당의 제조가 가능하며, 이를 과민성 장염 증후군의 예방, 개선 또는 치료용 약학 조성물, 건강기능식품으로 제공가능하다. The present invention relates to a method for producing isomaltooligosaccharide using rice flour, and by controlling the reaction time and reaction temperature of rice flour and various enzymes, it is possible to produce isomaltooligosaccharide with significantly enhanced prebiotic function than before, This can be provided as a pharmaceutical composition for the prevention, improvement or treatment of irritable bowel syndrome, or as a health functional food.
또한 본 발명에서 이용하는 쌀가루는 전분의 주원료로 사용하는 옥수수에 비하여 원료의 가격이 높은 면이 있지만, 옥수수의 전분을 별도로 생산하는 것에 비해 쌀을 분말화만 하면 되기에 생산단가가 낮고 공정상 보다 간단한 방법으로 쌀가루의 제조가 가능하여 옥수수를 이용하여 제조한 전분을 이용하여 이소말토올리고당을 제조함에 있어 비용/시간 면에서 더 유리하다. 게다가 전분의 주원료로 사용하는 옥수수의 경우 GMO 제품이 많은 반면, 쌀은 GMO 작물을 사용하지 않아 안전한 먹거리를 제공할 수 있다는 장점이 있다. In addition, the rice flour used in the present invention has a higher raw material price compared to corn used as the main raw material for starch, but compared to separately producing corn starch, the production cost is lower and the production cost is simpler because the rice flour only needs to be powdered. As it is possible to produce rice flour, it is more advantageous in terms of cost/time in manufacturing isomaltooligosaccharide using starch prepared from corn. In addition, while corn, which is used as the main raw material for starch, contains many GMO products, rice has the advantage of providing safe food because it does not use GMO crops.
도 1은 본 발명의 실시예 1에서 제조한 이소말토올리고당을 나타내는 사진이다. 1 is a photograph showing the isomaltooligosaccharide prepared in Example 1 of the present invention.
이하 본 발명의 바람직한 실시예를 상세히 설명하기로 한다. 그러나, 본 발명은 여기서 설명되는 실시예에 한정되지 않고 다른 형태로 구체화될 수도 있다. 오히려, 여기서 소개되는 내용이 철저하고 완전해지도록, 당업자에게 본 발명의 사상을 충분히 전달하기 위해 제공하는 것이다. Hereinafter, preferred embodiments of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein and may be embodied in other forms. Rather, it is provided so that this disclosure will be thorough and complete, and will fully convey the spirit of the invention to those skilled in the art.
<실시예 1. 단계별 효소처리를 통해 제조한 이소말토올리고당> <Example 1. Isomaltooligosaccharide prepared through step-by-step enzyme treatment>
쌀가루의 액화 반응을 위해서 쌀가루(백미)와 증류수를 1:4의 중량비로 하여 쌀가루 100g 당 증류수 400g을 혼합하여 분산시켰다. 당화 효소의 안정화를 위해 칼슘이온이 70ppm이 되도록 CaCO3 0.035g을 첨가하고 1N의 HCl를 사용하여 pH를 6.2로 조정하였다. For the liquefaction reaction of rice flour, 400 g of distilled water per 100 g of rice flour was mixed and dispersed by mixing rice flour (white rice) and distilled water in a weight ratio of 1:4. For stabilization of the glycosylation enzyme, 0.035 g of CaCO 3 was added so that the calcium ion was 70 ppm, and the pH was adjusted to 6.2 using 1N HCl.
이렇게 만들어진 실온 온도(25℃) 상태의 쌀가루 분산액을 60℃ 항온수조에 넣고 진탕하다가 쌀 분산액 온도가 60℃가 되면 쌀가루 자체의 급격한 호화를 방지하기 위해, 바실러스 리케니포르미스 균주로부터 생산된 α-Amylase(Termamyl 2X)를 첨가하였다. 다음으로는 95℃로 승온하여 2시간 동안 반응하게 하여 분산액 상태의 시료를 액화 상태로 제조하였다. In order to prevent rapid gelatinization of the rice flour itself when the temperature of the rice dispersion reaches 60 °C while shaking the resulting rice flour dispersion at room temperature (25 °C) in a constant temperature water bath at 60 °C, α- produced from Bacillus licheniformis strain Amylase (Termamyl 2X) was added. Next, the temperature was raised to 95° C. and reacted for 2 hours to prepare a sample in a dispersion state in a liquefied state.
이 때 바실러스 리케니포르미스 균주로부터 생산된 α-Amylase는 쌀가루 100 g 대비 0.075g을 사용하였다. At this time, α-Amylase produced from the Bacillus licheniformis strain was used in an amount of 0.075 g compared to 100 g of rice flour.
액화를 마친 액화액은 당화효소가 작용하는 최적 조건을 위해 55℃로 식혔고, pH도 1 N HCl 또는 1 N NaOH로 pH를 5.0으로 조정하였다.After liquefaction, the liquefied liquid was cooled to 55° C. for optimal conditions for the saccharification enzyme to act, and the pH was also adjusted to 5.0 with 1 N HCl or 1 N NaOH.
당화를 위해 바실러스 서브틸리스아밀로리퀘파시엔스 균주로부터 생산된 β-Amylase(Maltogenase L), 바실러스 애시도풀루리티커스 균주로부터 생산된 Pullulanase(Promozyme D2), 아스퍼질러스 니거 균주로부터 생산된 Transglucosidase(Transglucosidase L)를 쌀 액화액에 첨가하였는데, 최적 효소농도로서, 쌀가루 100 g 대비 β-Amylase(Maltogenase L) 0.0015g, Pullulanase(Promozyme) 0.07g, Transglucosidase(Transglucosidase L) 0.1g을 첨가하였다. 이 농도로 효소를 처리할 때, glucose의 함량은 감소되고 중합도가 높은 이소말토올리고당의 함량이 증가하는 효과가 있다. β-Amylase (Maltogenase L) produced from Bacillus subtilis amyloliquefaciens strain for saccharification, Pullulanase (Promozyme D2) produced from Bacillus acido pulluliticus strain, Transglucosidase produced from Aspergillus niger strain (Transglucosidase L) was added to the rice liquefied solution, and as the optimal enzyme concentration, 0.0015 g of β-Amylase (Maltogenase L), 0.07 g of Pullulanase (Promozyme), and 0.1 g of Transglucosidase (Transglucosidase L) compared to 100 g of rice flour were added. When the enzyme is treated at this concentration, the content of glucose is reduced and the content of isomaltooligosaccharide having a high degree of polymerization is increased.
이렇게 3종의 효소를 첨가하고 55℃를 유지하면서 36시간 반응시켜, 총 이소말토올리고당이 75.36 g/L을 포함하는 조성물을 얻었다. 최적 조건으로 제조된 이소말토올리고당은 18 brix 였고, isomaltose 35.11 g/L, panose 11.79 g/L, isomaltotriose 19.95 g/L, isomaltotetraose 7.46 g/L, isomaltopentaose 1.05 g/L 이 생성되었으며, 총당 중 이소말토올리고당의 함량은 56.37 중량%로 확인된다. In this way, three kinds of enzymes were added and reacted for 36 hours while maintaining 55° C., to obtain a composition containing 75.36 g/L of total isomaltooligosaccharide. The isomaltooligosaccharide prepared under optimal conditions was 18 brix, and isomaltose 35.11 g/L, panose 11.79 g/L, isomaltotriose 19.95 g/L, isomaltotetraose 7.46 g/L, and isomaltopentaose 1.05 g/L were produced. The content of the oligosaccharide is found to be 56.37 wt%.
이 과정에서 사용된 각 액화 및 당화를 위한 효소는 다음의 표 1과 같다.Enzymes for each liquefaction and saccharification used in this process are shown in Table 1 below.
(α-Amylase)Termamyl2X
(α-Amylase)
(Bacillus licheniformis)Bacillus licheniformis
( Bacillus licheniformis )
(β-Amylase)Maltogenase L
(β-Amylase)
서브틸리스아밀로리퀘파시엔스
(Bacillus subtilisamyloliquefaciens)bacillus
subtilis amyloliquefaciens
( Bacillus subtilisamyloliquefaciens )
(Pullulanase)PromozymeD2
(Pullulanase)
(Bacillus acidopullulyticus)Bacillus acidopuluriticus
( Bacillus acidopullulyticus )
(Transglucosidase)Transglucosidase L
(Transglucosidase)
(Aspergillus niger)Aspergillus niger
( Aspergillus niger )
β-Amylase(maltogenase L), Pullulanase(promozyme D2), Transglucosidase L 효소로 36시간의 당화 및 전이를 거친 쌀 당화액을 filter paper로 여과하였고, 여과된 당액을 진공농축기로 농축하여 75 brix의 당도로 제조하였다.The saccharified rice solution that had undergone saccharification and transfer for 36 hours with β-Amylase (maltogenase L), Pullulanase (promozyme D2), and Transglucosidase L enzymes was filtered with filter paper, and the filtered sugar solution was concentrated with a vacuum concentrator to a sugar content of 75 brix. prepared.
<실시예 2. 효소반응 시간을 조절하여 제조한 이소말토올리고당> <Example 2. Isomaltooligosaccharide prepared by controlling the enzyme reaction time>
실시예 2-1Example 2-1
실시예 1과 동일하게 당류 조성물을 제조하되, 95℃에서의 효소 반응 시간을 1시간으로 하였다. A saccharide composition was prepared in the same manner as in Example 1, except that the enzymatic reaction time at 95°C was 1 hour.
실시예 2-2Example 2-2
실시예 1과 동일하게 당류 조성물을 제조하되, 95℃에서의 효소 반응 시간을 3시간으로 하였다. A saccharide composition was prepared in the same manner as in Example 1, except that the enzymatic reaction time at 95°C was 3 hours.
실시예 2-3Example 2-3
실시예 1과 동일하게 당류 조성물을 제조하되, 3종 효소 반응시간을 30시간으로 하였다. A saccharide composition was prepared in the same manner as in Example 1, except that the reaction time for the three enzymes was 30 hours.
실시예 2-4Example 2-4
실시예 1과 동일하게 당류 조성물을 제조하되, 3종 효소 반응시간을 40시간으로 하였다. A saccharide composition was prepared in the same manner as in Example 1, except that the reaction time for the three enzymes was 40 hours.
<비교예 1. 각 효소의 종류를 조절하여 제조한 당류 조성물><Comparative Example 1. Sugar composition prepared by adjusting the type of each enzyme>
비교예 1-1 Comparative Example 1-1
실시예 1과 동일하게 이소말토올리고당 같은 당류 조성물을 제조하되, 첫 번째 효소처리 단계를 생략하고 바로 3종의 효소처리 단계를 수행하여 쌀가루 분산액을 당화하여 당류 조성물을 제조하였다. A saccharide composition such as isomaltooligosaccharide was prepared in the same manner as in Example 1, except that the first enzymatic treatment step was omitted and three enzymatic treatment steps were immediately performed to saccharify the rice flour dispersion to prepare a saccharide composition.
비교예 1-2Comparative Example 1-2
실시예 1과 동일하게 당류 조성물을 제조하되, 두 번째 3종의 효소처리 대신 바실러스 서브틸리스아밀로리퀘파시엔스 균주로부터 생산된 β-Amylase(Maltogenase L)만을 실시예 1의 3종 효소와 동일 중량으로 쌀 액화액에 처리하였다. A saccharide composition was prepared in the same manner as in Example 1, except for β-Amylase (Maltogenase L) produced from the Bacillus subtilis amyloliquefaciens strain instead of the second three kinds of enzyme treatment. Same as the three enzymes of Example 1. It was treated in the rice liquefied solution by weight.
비교예 1-3Comparative Example 1-3
실시예 1과 동일하게 당류 조성물을 제조하되, 두 번째 3종의 효소처리대신 바실러스 애시도풀루리티커스 균주로부터 생산된 Pullulanase(Promozyme D2)만을 실시예 1의 3종 효소와 동일 중량으로 쌀 액화액에 처리하였다. A saccharide composition was prepared in the same manner as in Example 1, but instead of the second three kinds of enzyme treatment, only Pullulanase (Promozyme D2) produced from the Bacillus acidopuluriticus strain was liquefied rice with the same weight as the three enzymes of Example 1. treated with the liquid.
비교예 1-4Comparative Example 1-4
실시예 1과 동일하게 당류 조성물을 제조하되, 두 번째 3종의 효소처리대신 아스퍼질러스 니거 균주로부터 생산된 Transglucosidase L만을 실시예 1의 3종 효소와 동일 중량으로 쌀 액화액에 처리하였다.A saccharide composition was prepared in the same manner as in Example 1, but only Transglucosidase L produced from the Aspergillus niger strain was treated with the same weight as the three enzymes of Example 1 in the rice liquefied solution instead of the second three kinds of enzyme treatment.
비교예 1-5Comparative Example 1-5
실시예 1과 동일하게 당류 조성물을 제조하되, 두 번째 3종의 효소처리대신 바실러스 서브틸리스아밀로리퀘파시엔스 균주로부터 생산된 β-Amylase(Maltogenase L), 바실러스 애시도풀루리티커스 균주로부터 생산된 Pullulanase(Promozyme D2) 2종 효소를 1:1의 중량비로 혼합하여 실시예 1의 3종 효소와 동일 중량으로 쌀 액화액에 처리하였다.A saccharide composition was prepared in the same manner as in Example 1, but instead of the second three kinds of enzyme treatment, β-Amylase (Maltogenase L) produced from the Bacillus subtilis amyloliquefaciens strain, The produced Pullulanase (Promozyme D2) two enzymes were mixed in a weight ratio of 1:1 and treated with the same weight as the three enzymes of Example 1 in the liquefied rice.
비교예 1-6Comparative Example 1-6
실시예 1과 동일하게 당류 조성물을 제조하되, 두 번째 3종의 효소처리대신 바실러스 서브틸리스아밀로리퀘파시엔스 균주로부터 생산된 β-Amylase(Maltogenase L), 아스퍼질러스 니거 균주로부터 생산된 Transglucosidase L 2종 효소를 1:1의 중량비로 혼합하여 실시예 1의 3종 효소와 동일 중량으로 쌀 액화액에 처리하였다.A saccharide composition was prepared in the same manner as in Example 1, but instead of the second three kinds of enzyme treatment, β-Amylase (Maltogenase L) produced from Bacillus subtilis amyloliquefaciens strain, Transglucosidase produced from Aspergillus niger strain L 2 enzymes were mixed in a weight ratio of 1:1 and treated with the same weight as the 3 enzymes of Example 1 in the liquefied rice.
비교예 1-7Comparative Example 1-7
실시예 1과 동일하게 당류 조성물을 제조하되, 두 번째 3종의 효소처리대신 바실러스 애시도풀루리티커스 균주로부터 생산된 Pullulanase(Promozyme D2), 아스퍼질러스 니거 균주로부터 생산된 Transglucosidase L 2종 효소를 1:1의 중량비로 혼합하여 실시예 1의 3종 효소와 동일 중량으로 쌀 액화액에 처리하였다.A saccharide composition was prepared in the same manner as in Example 1, except that, instead of treating the second three kinds of enzymes, Pullulanase (Promozyme D2) produced from Bacillus acidopuluriticus strain, Transglucosidase L two enzymes produced from Aspergillus niger strain was mixed in a weight ratio of 1:1 and treated with the rice liquefied solution at the same weight as the three enzymes of Example 1.
<비교예 1. 각 효소처리의 반응시간을 조절하여 제조한 이소말토올리고당><Comparative Example 1. Isomaltooligosaccharide prepared by controlling the reaction time of each enzyme treatment>
비교예 2-1Comparative Example 2-1
실시예 1과 동일하게 이소말토올리고당을 제조하되, 95℃에서의 효소 반응 시간을 4시간으로 하였다. Isomaltooligosaccharide was prepared in the same manner as in Example 1, but the enzymatic reaction time at 95°C was 4 hours.
비교예 2-2Comparative Example 2-2
실시예 1과 동일하게 이소말토올리고당을 제조하되, 95℃에서의 효소 반응 시간을 0.5시간으로 하였다. Isomaltooligosaccharide was prepared in the same manner as in Example 1, but the enzyme reaction time at 95°C was 0.5 hours.
비교예 2-3Comparative Example 2-3
실시예 1과 동일하게 이소말토올리고당을 제조하되, 3종 효소 반응시간을 20시간으로 하였다. Isomaltooligosaccharide was prepared in the same manner as in Example 1, but the reaction time of the three enzymes was 20 hours.
비교예 2-4Comparative Example 2-4
실시예 1과 동일하게 이소말토올리고당을 제조하되, 3종 효소 반응시간을 50시간으로 하였다. Isomaltooligosaccharide was prepared in the same manner as in Example 1, but the reaction time of the three enzymes was set to 50 hours.
비교예 2-5Comparative Example 2-5
실시예 1과 동일하게 이소말토올리고당을 제조하되, 첫 번째 효소처리 단계에서 승온을 55℃까지만 하고 90~100℃의 작용활성이 없는 Aspergillus niger 유래의 α-amylase(Fungamyl 800 L, 5.0-6.0, 53~58℃에서 작용함)를 넣어주고 55℃ 그대로를 2시간 동안 유지한 후, 두 번째 당화효소 처리단계를 진행하였다. Prepare isomaltooligosaccharide in the same manner as in Example 1, except that in the first enzyme treatment step, the temperature was raised up to 55 ° C and α-amylase derived from Aspergillus niger (Fungamyl 800 L, 5.0-6.0, 53~58°C) and maintained at 55°C for 2 hours, followed by a second saccharification enzyme treatment step.
※ 첫 번째 효소처리 단계만 수행하고 3종의 효소처리 단계를 생략하는 경우는 당화반응이 일어나지 않아 실험하지 않음. 또한 첫 번째 효소처리 단계에서 바실러스 리케니포르미스 균주로부터 생산된 α-Amylase를 60℃에서 처리하지 않고 95℃까지 승온 후 바로 첨가하는 과정도 쌀가루의 응집현상이 일어나 다음의 당화과정을 진행할 수 없어 실험하지 않음. ※ If only the first enzymatic treatment step is performed and the 3 enzymatic treatment steps are omitted, the saccharification reaction does not occur and the experiment is not performed. In addition, in the process of adding α-Amylase produced from the Bacillus licheniformis strain in the first enzyme treatment step immediately after raising the temperature to 95°C without treatment at 60°C, aggregation of rice flour occurs, so the next saccharification process cannot proceed. Not tested.
<실험예 1. 속박 스트레스에 의한 배변 모델(Restraint stress-induced fecal pellet output model)에서의 효과 평가><Experimental Example 1. Effect evaluation in the constraint stress-induced fecal pellet output model>
실시예 1 및 비교예 1의 이소말토올리고당 또는 당류의 과민성 대장 증후군 개선 기능을 속박 스트레스에 의한 배변 모델(Restraint stress-induced fecal pellet output model)[S. Kobayashi 외, Jpn. J. Pharmcaol., 86, p 281-288, 2001]을 이용하여 평가하였다. Example 1 and Comparative Example 1 of isomaltooligosaccharide or saccharides of irritable bowel syndrome improvement function by constraint stress defecation model (Restraint stress-induced fecal pellet output model) [S. Kobayashi et al., Jpn. J. Pharmcaol., 86, p 281-288, 2001].
실험 동물로는 체중 250~300g의 Sprague-Dawley rat(Charles River) 수컷을 이용하였고, 상기 SD rat을 온도 25℃, 습도 50%, 낮-밤 사이클 12:12시간으로 조절된 동물실에서 케이지 당 두 마리씩 사육하였다. 물과 사료는 자유롭게 접근할 수 있도록 하였으며 5일간 적응시킨 후 속박 실험을 하였다.As experimental animals, male Sprague-Dawley rats (Charles River) weighing 250-300 g were used, and the SD rats were subjected to a temperature of 25° C., humidity 50%, and a day-night cycle of 12:12 hours in an animal room controlled for 12 hours per cage. Two were bred. Water and feed were freely accessible, and after 5 days of acclimatization, a restraint experiment was performed.
실험 당일, 보정틀(restraint cage)을 사용하여 SD rat에서 속박 스트레스에 의한 배변 양상(Restraint-induced fecal pellet output)을 측정하였다. 이를 위해 각각의 실시예 1 및 비교예 2의 각 조성물을 0.5%(w.v) CMC 수용액에 녹여 300 mg/kg의 농도로 경구투여하고 실험동물을 보정틀에 집어넣었다. 이때 동물이 투여로 인한 스트레스를 받지 않도록 조심하였다. 보정틀 안에서 움직이지 못하게 되면 동물은 속박 스트레스를 받게 되고 배변 활동을 시작하게 된다.On the day of the experiment, restraint-induced fecal pellet output was measured in SD rats using a restraint cage. To this end, each composition of Examples 1 and 2 was dissolved in 0.5% (w.v) CMC aqueous solution and orally administered at a concentration of 300 mg/kg, and the experimental animals were placed in a calibration frame. At this time, care was taken not to stress the animals due to administration. Immobility within the cage puts the animal under bondage stress and initiates defecation.
변의 양상과 개수를 60분 간격으로 4시간 동안 측정하였으며, 그 결과를 표 2에 나타내었다. 통계처리는 Student's t-test를 이용하여 p<0.01 (**) 또는 p<0.001 (***) 수준에서 유의성을 검정하였다.The pattern and number of stools were measured at 60-minute intervals for 4 hours, and the results are shown in Table 2. For statistical treatment, significance was tested at the level of p<0.01 (**) or p<0.001 (***) using Student's t-test.
그 결과 상기 표 2와 같이 본 발명의 이소말토올리고당을 처리한 군은 속박스트레스로 인해 발생하는 배변 횟수(fecal pellet output)가 이소말토올리고당 무처리군(물) 대비 현저하게 감소하였고, 속박스트레스를 전혀 가하지 않은 일반군과 유사하게 변을 배출하는 것으로 확인되었다. 변의 상태 및 복부팽만 상태도 비교한 바, 본 발명의 이소말토올리고당이 투여된 군은 일반군과 유사하게 단단하였으나, 비교예 1의 당류가 투여된 군은 대부분 설사로 체크되었고, 복부 팽만 상태도 심각하였다. 특히 상기 이 결과를 통해 조청이나 올리고당, 시판 이소말토올리고당 자체만으로는 복부팽만 상태나 설사나 잦은 배변횟수를 줄이는 데에 거의 영향을 주지 않는 것을 알 수 있다. As a result, as shown in Table 2 above, in the group treated with isomaltooligosaccharide of the present invention, the number of bowel movements (fecal pellet output) caused by constraint stress was significantly reduced compared to the isomaltooligosaccharide untreated group (water), and the constraint stress was reduced. It was confirmed that feces were excreted similarly to the general group, which was not applied at all. When the state of stool and distension of the abdomen were also compared, the group administered with the isomaltooligosaccharide of the present invention was hard similar to the general group, but the group administered with the saccharide of Comparative Example 1 was mostly checked for diarrhea, and the state of abdominal distension was also It was serious. In particular, from these results, it can be seen that crude oil, oligosaccharides, and commercially available isomaltooligosaccharides alone have little effect on reducing abdominal distension, diarrhea, or frequent bowel movements.
<실험예 2. 장내 균의 배양 효능 확인> <Experimental Example 2. Confirmation of the culture efficacy of intestinal bacteria>
실험예 1에서 4시간 동안 SD rat에서 얻은 분변을 각 처리군별로 모두 모아 이 분변 1g을 생리식염수를 첨가하여 총 10㎖이 되도록 희석한 것을 실험용 원료 시료로 사용하였다. 각 시료를 다시 10배 희석법으로 희석하여 얻은 액상을 유산균 배양용 배지인 MRS 아가 배지에 37℃에서 20시간씩 배양하여 생성되는 colony 수를 세어 최초 분변의 1g/10㎖ 희석액 내 총 유산균 수를 측정하였다. 분변 내 총 유산균 균수를 속박 스트레스 비처리군을 100의 값으로 기준하여 다음의 표 3에 대비하여 기재하였다. In Experimental Example 1, all feces obtained from SD rats for 4 hours were collected for each treatment group, and 1 g of the feces was diluted to a total of 10 ml by adding physiological saline, which was used as a raw material sample for the experiment. The liquid obtained by diluting each sample again by a 10-fold dilution method is cultured for 20 hours at 37°C in MRS agar medium, a medium for culturing lactic acid bacteria, and the number of colonies generated is counted. did. The total number of lactic acid bacteria in the feces was described in comparison with Table 3 below, based on a value of 100 for the untreated group under restraint stress.
(fold)Lactobacillus in 1g/10ml dilution of feces
(fold)
상기 표 3의 결과는, 속박 스트레스에 따른 배변 이상을 본 발명의 이소말토올리고당이 장내 유산균 수를 증강 또는 회복시켜 유용 미생물총을 개선하는 것을 뒷받침한다. 따라서 이 결과를 통해 본 발명에서 제조한 이소말토올리고당이 장내 프로바이오틱스의 증식을 조절하는 프리바이오틱스로서 이용되기에 적합한 조성물인 것을 확인할 수 있다. 또한 상기 결과를 통해 시판 이소말토올리고당이나 올리고당, 조청 만으로는 유산균 총균수의 조절이 잘 되지 않음을 알 수 있다. The results of Table 3 support the improvement of the useful microflora by enhancing or restoring the number of intestinal lactic acid bacteria by the isomaltooligosaccharide of the present invention for defecation abnormalities according to restraint stress. Therefore, through this result, it can be confirmed that the isomaltooligosaccharide prepared in the present invention is a composition suitable for use as a prebiotic for regulating the proliferation of intestinal probiotics. In addition, it can be seen from the above results that the control of the total number of lactic acid bacteria is not well with only commercially available isomaltooligosaccharides, oligosaccharides, and crude oil.
<제제예 1. 약학적 제제><Formulation Example 1. Pharmaceutical formulation>
제제예 1-1. 분말제형의 제조Formulation Example 1-1. Preparation of powder formulations
본 발명의 실시예 1의 이소말토올리고당을 건조하고 분쇄하여 분말화하고 동결건조된 유산균과 혼합하였다. The isomaltooligosaccharide of Example 1 of the present invention was dried, pulverized, powdered, and mixed with lyophilized lactic acid bacteria.
<제제예 2. 식품 제조><Formulation Example 2. Food Manufacturing>
제제예 2-1. 조리용 양념의 제조Formulation Example 2-1. Preparation of cooking seasonings
본 발명의 실시예 1의 이소말토올리고당을 조리용 양념에 1 중량%로 첨가하여 건강 증진용 조리용 양념을 제조하였다.A cooking seasoning for health promotion was prepared by adding the isomaltooligosaccharide of Example 1 of the present invention to the cooking seasoning in an amount of 1% by weight.
제제예 2-2. 밀가루 식품의 제조Formulation Example 2-2. Flour food manufacturing
본 발명의 실시예 1의 이소말토올리고당을 밀가루에 0.1 중량%로 첨가하고, 이 혼합물을 이용하여 빵, 케이크, 쿠키, 크래커 및 면류를 제조하여 건강 증진용 식품을 제조하였다.The isomaltooligosaccharide of Example 1 of the present invention was added to wheat flour in an amount of 0.1 wt%, and bread, cake, cookies, crackers and noodles were prepared using this mixture to prepare health-promoting foods.
제제예 2-3. 스프 및 육즙(gravies)의 제조Formulation Example 2-3. Preparation of soups and gravies
본 발명의 실시예 1의 이소말토올리고당을 스프 및 육즙에 0.1 중량%로 첨가하여 건강 증진용 수프 및 육즙을 제조하였다.The isomaltooligosaccharide of Example 1 of the present invention was added to the soup and broth in an amount of 0.1% by weight to prepare a health-improving soup and broth.
제제예 2-4. 유제품(dairy products)의 제조Formulation Example 2-4. Manufacture of dairy products
본 발명의 실시예 1의 이소말토올리고당을 우유에 0.1 중량%로 첨가하고, 상기 우유를 이용하여 버터 및 아이스크림과 같은 다양한 유제품을 제조하였다.The isomaltooligosaccharide of Example 1 of the present invention was added to milk in an amount of 0.1% by weight, and various dairy products such as butter and ice cream were prepared using the milk.
제제예 2-5. 야채주스 제조Formulation Example 2-5. vegetable juice production
본 발명의 실시예 1의 이소말토올리고당 0.5g을 토마토주스 또는 당근주스 1,000㎖에 가하여 건강 증진용 야채주스를 제조하였다.0.5 g of isomaltooligosaccharide of Example 1 of the present invention was added to 1,000 ml of tomato juice or carrot juice to prepare vegetable juice for health promotion.
제제예 2-6. 과일주스 제조Formulation Example 2-6. fruit juice manufacturing
본 발명의 실시예 1-2의 이소말토올리고당을 0.1g을 사과주스 또는 포도주스 1,000㎖에 가하여 건강 증진용 과일주스를 제조하였다.Fruit juice for health promotion was prepared by adding 0.1 g of the isomaltooligosaccharide of Example 1-2 of the present invention to 1,000 ml of apple juice or grape juice.
Claims (9)
(제2단계) 상기 쌀가루 슬러리를 승온시켜 58~62℃에서 α-아밀라아제를 첨가하고, 상기 효소 첨가 후 쌀가루 슬러리의 온도를 계속 승온시켜 90~95℃로 온도를 유지한 상태에서 효소 반응하여 쌀가루 슬러리를 액화하는 단계;
(제3단계) 제2단계에서 얻은 쌀가루 액화액의 온도를 55~60℃로 하강하여 상기 온도를 유지한 상태에서, β-아밀레아제, 풀루라나아제 및 트랜스글루코시다아제를 쌀 액화액에 첨가하고 반응하여 상기 액화액을 당화함으로써 이소말토올리고당을 얻는 단계;
(제4단계) 제3단계에서 얻은 이소말토올리고당을 여과 및 농축하여 70~80 브릭스 당도의 이소말토올리고당을 얻는 단계;
를 포함하는 것을 특징으로 하는 쌀가루를 이용한 이소말토올리고당의 제조방법. (Step 1) preparing a rice flour slurry by adding CaCO 3 to the rice flour dispersion so that calcium ions become 60-80 ppm, and adjusting the pH to 6.0-6.4 while adding 0.8-1.2N HCl dropwise;
(Second step) The temperature of the rice flour slurry is raised and α-amylase is added at 58 to 62 ° C. After the enzyme is added, the temperature of the rice flour slurry is continuously raised to maintain the temperature at 90 to 95 ° C. liquefying the slurry;
(3rd step) β-amylase, pullulanase and transglucosidase were added to the rice liquefied solution while maintaining the temperature by lowering the temperature of the rice flour liquefied liquid obtained in the second step to 55-60 ° C. adding and reacting to saccharify the liquid to obtain isomaltooligosaccharide;
(Step 4) filtering and concentrating the isomaltooligosaccharide obtained in step 3 to obtain isomaltooligosaccharide having a sugar content of 70 to 80 Brix;
Method for producing isomaltooligosaccharide using rice flour, characterized in that it comprises a.
상기 제3단계에서 얻은 이소말토올리고당이 16~18 브릭스의 당도를 갖는 것을 특징으로 하는 쌀가루를 이용한 이소말토올리고당의 제조방법.According to claim 1,
A method for producing isomaltooligosaccharide using rice flour, characterized in that the isomaltooligosaccharide obtained in the third step has a sugar content of 16 to 18 brix.
상기 제3단계에서 얻은 이소말토올리고당이 이소말토오스 30~40 g/L, 판노스 8~15 g/L, 이소말토트리오스 15~25 g/L, 이소말토테트라오스(5~10 g/L, 이소말토펜타오스 0.5~2 g/L를 포함하고, 총 이소말토올리고당이 60~95g/L인 것을 특징으로 하는 쌀가루를 이용한 이소말토올리고당의 제조방법. According to claim 1,
The isomaltooligosaccharide obtained in the third step is isomaltose 30-40 g/L, pannose 8-15 g/L, isomaltotriose 15-25 g/L, isomaltotetraose (5-10 g/L) , A method for producing isomaltooligosaccharide using rice flour, characterized in that it contains 0.5-2 g/L of isomaltopentaose, and the total isomaltooligosaccharide is 60-95g/L.
상기 제3단계에서 얻은 이소말토올리고당을 함유한 조성물 내의 총 당류 100 중량% 기준, 이소말토올리고당의 함량이 50~60 중량%인 것을 특징으로 하는 쌀가루를 이용한 이소말토올리고당의 제조방법. According to claim 1,
A method for producing isomaltooligosaccharide using rice flour, characterized in that the content of isomaltooligosaccharide is 50 to 60% by weight based on 100% by weight of total saccharides in the composition containing isomaltooligosaccharide obtained in the third step.
상기 제4단계에서 농축 후 얻은 이소말토올리고당의 당도가 70~80 브릭스인 것을 특징으로 하는 이소말토올리고당의 제조방법. According to claim 1,
The method for producing isomaltooligosaccharide, characterized in that the sugar content of the isomaltooligosaccharide obtained after concentration in the fourth step is 70-80 brix.
상기 제4단계 이후, (제5단계) 상기 조성물을 분말화하여 분말을 얻는 단계;가 더 포함되는 것을 특징으로 하는 이소말토올리고당의 제조방법. According to claim 1,
After the fourth step, (fifth step) pulverizing the composition to obtain a powder; Method for producing isomaltooligosaccharide, characterized in that it further comprises.
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