WO2013081294A1 - 싸이코스를 유효성분으로 하는 에쿠올 농도 상승제 - Google Patents
싸이코스를 유효성분으로 하는 에쿠올 농도 상승제 Download PDFInfo
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- WO2013081294A1 WO2013081294A1 PCT/KR2012/008398 KR2012008398W WO2013081294A1 WO 2013081294 A1 WO2013081294 A1 WO 2013081294A1 KR 2012008398 W KR2012008398 W KR 2012008398W WO 2013081294 A1 WO2013081294 A1 WO 2013081294A1
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Definitions
- the present invention relates to an equol concentration increasing agent having sicose as an active ingredient, and more particularly, to an equol concentration increasing agent having sicose as an active ingredient in order to promote the production of equol by human intestinal microorganisms. will be.
- Equol (Iquol) is an isoflavan compound having the structure of Formula 1 below, when ingesting legumes food is produced by the conversion of the intestinal microorganisms of isoflavonoids present in the food.
- Equol the active metabolite of soy isoflavones, is reported to hold the key to the effectiveness of soy foods in clinical applications.
- equol which is a metabolite, is more effective than soy isoflavones for breast cancer, prostate cancer, anti-aging, menopausal disorders and postmenopausal osteoporosis
- equol is produced by intestinal bacteria, and it has been reported that individual differences exist in the production of Nutr Cancer. 2000; 36: 27-32. Those who cannot produce equol are estimated to have no equol-producing bacteria in their gut. In such a person, the anti-estrogen effect and the estrogen-like effect that the processed soybeans are eaten may not be expected. In order to express the desired effect in such a person, it is thought that the equol-producing microorganisms may be ingested or the equol itself may be ingested.
- sugar alcohols which are widely used as sugar substitute sweeteners, cause side effects, such as diarrhea, when consumed over a certain amount, whereas psychos have little side effects (Matsue, T., Y. Baba, M. Hashiguchi, K. Takeshita, K. Izumori, and H. Suzuki.2001.Dietary D-psicose, a C-3 epimer of D-fructose, suppresses the activity of hepatic lipogenic enzymes in rats.Asia Pac.J. Clin.Nutr. 10 Matsuo, T., and K. Izumori. 2004. D-psicose, a rare sugar that provides no energy and additionally beneficial effects for clinical nutrition.Asia Pac.J. Clin.Nutr. 13: S127. ).
- the present inventors have sought to find a functional food having an effect of promoting the conversion of equol in addition to the general saccharides and polysaccharides reported to date.
- the present invention has led to a significant result that cyclose, known as a functional sweetener, unexpectedly promotes the production of equol, thereby completing the present invention.
- the primary purpose of the screening was to select a material showing a high conversion promoting rate based on the existing literature. If only the materials with high equol conversion promoting rate of the existing papers were selected, lactitol, inositol, melezitoose, sorbose, trehalose and xylose were included.
- xylose was selected as a representative control material, and in order to minimize inaccuracies in the difference between experimental methods and the presence or absence of experimental errors, it was relatively more effective than xylose.
- sugars with high conversion were intended to be defined as test substances with the effect of promoting equol conversion.
- the researchers conducted a series of experiments on new generic (functional) saccharides that had not been previously evaluated that met this goal, among which the results showed significant results that psychos promoted equol production. The present invention has been completed.
- the present invention relates to an equol concentration increasing agent having sicose as an active ingredient, and more particularly to an equol concentration increasing agent having sicose as an active ingredient in order to promote the production of equol by human intestinal microorganisms. It is about.
- the main object of the present invention is to provide a functional food having the effect of further promoting the conversion of equol in addition to the general saccharides and polysaccharides reported to date.
- Another object of the present invention is to provide a new use of Psicose, one of the rare saccharides.
- Still another object of the present invention is to provide a composition or food having a function of activating equol conversion and having a safe and physiological function activating effect without side effects when administered to the human body.
- the present invention has found that a safe and low-calorie functional sweetener, as a food, has a function of increasing equol concentration and has completed the present invention.
- a medium containing cyclose has a function of selectively propagating microorganisms having equol converting ability or enhancing equol converting ability.
- the present invention provides a composition or food having a function of safe and physiological function activating action and promoting equol conversion when administered to the human body.
- 3 is a graph showing the effect of various sugars on the equol change.
- the cosmos used in the present invention refers to D-psicose, which is contained in a small amount of molasses or isosaccharide, and is a kind of rare sugar that can be enzymatically produced by epimerase from D-fructose.
- the psychos are expected to have physiological functions such as antioxidant properties, and even though the sweetness reaches 60-70% of the sugar, the energy is almost O kilocalories and has excellent solubility.
- One aspect of the present invention relates to an equol concentration increasing agent using the aforementioned sicose (Psicose) as an active ingredient.
- the synergistic effect of the equol concentration of the psychos is due to the selective growth of the microorganisms having the equol converting ability or the increase of the equol converting ability of the microorganism.
- equol concentration-increasing agent in the intestine of the body, blood, large intestine, etc. It can be used for the purpose of treating and improving various diseases related to isoflavones, such as menopausal disorders, osteoporosis, hyperlipidemia, arteriosclerosis, breast cancer, prostate cancer, and premenstrual syndrome, or the like.
- equol producing bacterium a method of fermenting with a microorganism (hereinafter referred to as equol producing bacterium) that metabolizes diedzein to produce equol with respect to a raw material containing diedzein Known. Therefore, it is preferable to use the cyclos of this invention with the dyedzein which is a substrate of an equol.
- Dyed zein may use commercially available products, such as a synthetic product and a natural extract, and can use the natural material which contains a large amount of dyed zein, and its processed goods.
- materials containing a lot of dyed zein include soybeans, peas, sesame seeds, clover, and the like, and their processed products include tofu, soy milk, tempura, sickle, soy sauce, miso, and tempe.
- isoflavone glycosides are generally aglyconized by the action of enterobacteriaceae in the body, diedzein can also be used in the form of its glycoside compounds, such as dydazine, malonididezin, and acetyldidezin.
- Known equol-producing strains can be used without particular limitation in the present invention, for example, Bacteroides Ovartus, Streptococcus intermediars, Streptococcus consterus, Lactococcus garvies and the like. More specifically, Bacteroides E-23-15 (FERM BP-6435), Streptococcus E-23-17 (FERM BP-6436), Streptococcus A6G225 (FERM BP-6437), Lactoko Cous 20-92 (FERM BP-10036).
- the present invention can increase the equol production concentration by fermenting the equol producing bacteria by adding the cosmos to the medium in the conventional equol production method described above.
- the equol concentration increasing agent according to the present invention may further include other saccharides known to have an effect of raising the equol concentration in addition to the cyclos.
- Other sugars that may be added are, for example, adonitol, arabinose, erythritol, galactose, lactitol, melezitose, trehalose, ribose, sorbose, xylose, inositol Or sorbitol, and the like, but is not limited thereto.
- another aspect of the present invention relates to a method for increasing equol concentration, which comprises adding cyclose to a culture medium of an equol producing strain.
- Another aspect of the present invention is to provide a culture medium of the equol-producing strain, characterized in that it contains a cycle.
- Yet another aspect of the present invention relates to a food for increasing equol concentration, which includes a picos.
- the difference in the amount of equol produced according to the type of carbohydrate was identified using a group of microorganisms, which is a collection of various intestinal microorganisms.
- a group of microorganisms which is a collection of various intestinal microorganisms.
- carbohydrates affected the change of the microflora and the growth of equol-producing microorganisms (Example 2).
- the reaction rate of the amount of equol produced remained when carbohydrates were added, unlike when pure Dyzedine was used. That is, the production of equol takes place more slowly over a longer period of time.
- Cycose equol concentration regulating activity is Dyzezein, which is a substrate of equol, in microbial group stock (Central washing of feces collected from equol producer, KR registration 2008-0072635) having equol-producing ability, and The culture was carried out by adding cyclose, and the concentration of equol was measured by measuring the concentration of equol in the culture solution and comparing the concentration with equol in the culture solution containing dydzein and xylose.
- the dosage of the equol concentration increasing agent according to the present invention is not strict, and since the effect obtained varies depending on various types of use, such as the subject or the applied disease, it is preferable to set an appropriate dosage, and the dosage is per day. 0.1 mg to 100 g, with 50 mg to 50 g being particularly preferred.
- the equol concentration estimating agent of the present invention can be used either orally or parenterally, but oral administration is preferred. At the time of administration, it can be administered in the form of a conventional pharmaceutical formulation by mixing the active ingredient psychos with a solid or liquid medicinal nontoxic carrier suitable for oral administration, rectal administration, injection and the like.
- Such preparations include solid preparations such as tablets, granules, powders and capsules, solutions such as solutions, suspensions and emulsions, and lyophilizers. These formulations can be prepared by conventional methods on the formulation.
- the pharmaceutical non-toxic carriers include starch, dextrin, fatty acid glycerides, polyethylene glycol, hydroxyethylene starch, ethylene glycol, polyoxyethylene sorbitan fatty acid esters, amino acids, gelatin, albumin, water, physiological saline and the like.
- general additives such as a stabilizer, a wetting agent, an emulsifier, a binder, an isotonicity agent, an excipient, can be added suitably as needed.
- the saccharide of the present invention can be used not only as a pharmaceutical preparation as described above, but also as a food or drink.
- the sugar of the present invention may be added as it is or in various food ingredients with various nutrients.
- the purpose is to increase the equol concentration in the body or to improve and prevent menopausal disorders such as negative hydrogen, osteoporosis, hyperlipidemia, arteriosclerosis, breast cancer, prostate cancer, and premenstrual syndrome. It can be used as health food or food material useful for the back.
- the additives that can be used as food or drink are suitably used for edible use, i.e., granular, granular, tablet, capsule, paste, or the like. It may be molded and used in addition to various foods such as meat processed foods such as ham and sausage, fish processed foods such as fish paste and oden, bread, confectionery, butter, powdered milk and fermented milk products, or used in water, fruit juice and milk. You may use it in addition to beverages, such as a soft drink and a tea drink. Food and drink also includes animal feed.
- DHD dihydrodyne zein
- Example 1 Activation of equol-producing microorganisms and measurement of equol production under standard conditions
- Centrifugal cleaning of equol-producing microorganism stocks stored in liquid nitrogen was inoculated in GAM ( Gifu anaerobic medium) medium (4 ml) for 3 to 4 hours. Raised and activated.
- a total of six carbohydrates such as xylose, tagatose, sicose, sucrose, fructooligosaccharide, and glucose, were used to measure the amount of equol production under the equol production conditions used in Example 1.
- Example 2 The same method as Example 1 was inoculated with the equol-producing microbial group and the product was extracted, and the difference was Xylose, tagatose, cyclose, sucrose, fructooligosaccharide, glucose before the substrate inoculation of the activated microbial group, respectively. Add 2mg of the medium to the final carbohydrate concentration to adjust to 1%. In addition, only Dyzedine was used as a substrate. Unlike the case of using pure Dyzedein, when the carbohydrate was added, there was a difference in the reaction rate of the amount of equol produced. In other words, the production of equol occurs more slowly over a longer period of time.
- the comparison result is as follows.
- the relative amount of equol produced by Xylose was compared with that of xylose (%), which shows the average value of the image analysis results for the two color development methods).
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Abstract
본 발명은 인체 장내 미생물에 의한 다이드제인의 에쿠올로의 전환을 촉진하는 기능을 가지는 당류인 싸이코스를 유효성분으로 하는 가공식품 조성물에 대한 것이다.
Description
본 발명은 싸이코스(Psicose)을 유효성분으로 하는 에쿠올 농도 상승제에 관한 것으로, 구체적으로 인체 장내 미생물에 의한 에쿠올 생성을 촉진하기 위해 싸이코스를 유효성분으로 하는 에쿠올 농도 상승제에 관한 것이다.
에쿠올(Equol)은 아래 화학식 1의 구조를 갖는 이소플라반 화합물로, 콩류 식품을 섭취하였을 때, 식품에 존재하는 이소플라보노이드의 장내 미생물에 의한 전환으로 생성된다.
[화학식 1]
대두 이소플라본의 활성 대사물인 에쿠올이 임상 응용에서의 대두 식품의 유효성의 열쇠를 쥐고 있는 것으로 보고되고 있다. 즉, 유방암, 전립선암, 노화 방지, 갱년기 장해 및 폐경 후의 골다공증에 대하여, 대두 이소플라본보다도 그 대사물인 에쿠올이 유효하다는 것을 의미하는 보고가 다수 발견된다(문헌[D. Ingram, et al., (1997) Lancet, 350, 990-994]; 문헌[A. M. Duncan, et al., (2000) Cancer Epidemiology, Biomarkers&Prevention, 9, 581-586]; 문헌[C. Atkinson, et al., (2002) J. Nutr., 32(3), 595S]; 문헌[H. Akaza, et al., (2002) Jpn. J. Clin. Oncol., 32(8), 296-300]; 문헌[S.Uchiyama,. et al., (2001) Ann. Nutr. Metab., 45, 113(abs); Biol Reprod. 2004;70:1188-95 등 참조).
또한, 에쿠올은 장내 세균에 의해서 생성되고, 그 생성에는 개인차가 존재하는 것으로 보고되어 있다 [문헌: Nutr Cancer. 2000;36:27-32]. 에쿠올을 생산할 수 없는 사람은 에쿠올 생산 생균이 장내에 존재하지 않는 것으로 추정된다. 이러한 사람의 경우, 대두 가공 식품을 섭취하여도 원하는 항 에스트로겐 효과, 에스트로겐 유사 효과는 기대할 수 없는 것으로 생각된다. 이러한 사람에게서 원하는 효과를 발현시키기 위해서는, 에쿠올 생산 생균을 섭취시키거나, 에쿠올 자체를 섭취시키면 좋다고 생각된다.
한편, 이렇듯 유용한 에쿠올의 생명공학적 합성에 대한 시도도 예상되나 에쿠올의 생성 반응이 여러 미생물을 필요로 하는 복잡한 생화학적 경로로 이루어져 있으며 (도 2 참조), 대부부의 혐기성 미생물과 마찬가지로 알려진 에쿠올 생성 미생물의 성장과 반응 속도가 매우 느린 점이 기술적으로 극복해야 할 과제로 남아 있다.
따라서 보다 실질적인 연구를 목적으로, 에쿠올 생산능을 가진 장내 세균이나 에쿠올 생산을 촉진하는 물질의 탐색에 많은 연구가 이루어지고 있으며, 에쿠올 생산능을 갖는 미생물로서 박테로이데스 오바터스, 스트렙토코카스 인터메디어스, 스트렙토코카스 콘스테라터스가 보고되어 있다 (국제 특허 출원 공개 제1999/07392 호 참조). 또한 다양한 선행연구를 통하여 각종 단당류 혹은 다당류가 우수한 에쿠올 농도의 상승 또는 저하작용을 가짐을 보고하였는데, 특히 아도니톨, 아라비노오즈, 에리스리톨, 갈락토오즈, 락티톨, 멜레지토오즈, 트레할로오즈, 리보오즈, 소르보오즈, 자일로스, 이노시톨 및 솔비톨 등의 당류가 에쿠올 농도 상승에 기여하는 것으로 보고되었다 (국제 특허 출원 공개 제2007/052749호의 아래 표 1 참조).
표 1
당의 종류 | 배지중의 에쿠올 농도(μM) | 에쿠올 농도 조절활성(%) |
아도니톨 | 47.0 | 260.0 |
아라비노오즈 | 70.6 | 391.1 |
셀로비오스 | 29.0 | 160.6 |
에리스리톨 | 38.7 | 214.2 |
프락토올리고당 | 0.0 | 0.0 |
프락토오즈 | 9.3 | 51.5 |
갈락토오즈 | 65.0 | 240.1 |
글루코오즈 | 0.0 | 0.0 |
글리코겐 | 33.4 | 184.8 |
이눌린 | 0.0 | 0.0 |
락티톨 | 82.9 | 459.1 |
락토오즈 | 0.0 | 0.0 |
락투로오즈 | 1.5 | 8.1 |
이노시톨 | 75.2 | 416.2 |
말토오즈 | 10.6 | 58.5 |
만니톨 | 32.9 | 182.2 |
만노오즈 | 15.0 | 83.0 |
멜레지토오즈 | 74.3 | 411.6 |
메리비오스 | 4.5 | 24.8 |
라피노오즈 | 0.0 | 0.0 |
람노오즈 | 21.0 | 116.4 |
리보오즈 | 59.5 | 329.6 |
소르비톨 | 67.3 | 372.5 |
소르보오즈 | 84.5 | 467.7 |
수크로오즈 | 0.0 | 0.0 |
갈락토올리고당 | 0.0 | 0.0 |
트레할로오즈 | 72.9 | 403.5 |
자일로스 | 79.9 | 442.6 |
당 비첨가 | 18.1 | 100.0 |
한편, 싸이코스는 설탕과 유사한 단맛을 가지나, 초저열량인 단당류로서 기능성 감미료로 널리 이용되고 있다. 특히, 싸이코스는 자연계에 극히 소량 존재하는 희귀당으로 keto-hexose로 분류된다. 싸이코스는 D-tagatose 3-epimerase 에 의해 fructose로부터 만들어진다고 보고되었다. 감미의 강도와 종류는 과당과 매우 유사하나, 과당과 달리 체내 흡수시 거의 대사되지 않기 때문에 열량은 제로에 가깝고 지질합성에 관여하는 효소 활성을 억제해서 복부 비만을 감소시키는 기능을 갖기 때문에, 다이어트 식품의 유효 성분으로 유용하게 사용될 수 있다. 또한, 설탕 대체 감미료로 널리 이용되고 있는 당알코올류는 일정량 이상 섭취시 설사를 유발하는 등의 부작용을 일으키는 반면, 싸이코스는 부작용이 거의 없다(Matsue, T., Y. Baba, M. Hashiguchi, K. Takeshita, K. Izumori, and H. Suzuki. 2001. Dietary D-psicose, a C-3 epimer of D-fructose, suppresses the activity of hepatic lipogenic enzymes in rats. Asia Pac. J. Clin. Nutr. 10:233-237.; Matsuo, T., and K. Izumori. 2004. D-psicose, a rare sugar that provides no energy and additionally beneficial effects for clinical nutrition. Asia Pac. J. Clin. Nutr. 13:S127 참조).
이에, 본 발명자들은 현재까지 보고된 일반 당류 및 다당류 외에 추가적으로 에쿠올의 전환을 촉진하는 효능을 가지는 기능성 식품을 발굴하고자 하였다. 예의노력을 거듭한 결과, 기능성 감미료로 알려진 싸이코스가 예상외로 에쿠올 생성을 촉진한다는 유의한 결과를 도출하고, 이로부터 본 발명을 완성하게 되었다.
다양한 당류의 에쿠올 전환 촉진 효능의 상대적 비교 평가를 하기 위하여, 1차적으로 기존 문헌을 바탕으로 높은 전환 촉진률을 보이는 물질을 선별하고자 하였다. 기존 논문의 에쿠올 전환 촉진율이 우수한 물질만 선별하면, 락티톨, 이노시톨, 멜레지토오즈, 소르보오즈, 트레할로스, 자일로스가 이에 해당하였다.
또한, 본 발명자는 당알콜류가 아닌 천연 유래의 일반 당류에 주목하였으므로, 대표적인 대조 물질로 자일로스를 선정하였으며, 발명자 간의 실험 방법 차이 및 실험적 오차 유무에 대한 부정확성을 최소화하기 위하여, 자일로스보다 상대적으로 높은 전환율을 보이는 당류만을 에쿠올 전환율을 촉진하는 효과가 있는 시험 물질로 규정하고자 하였다. 연구자들은 이러한 목적에 부합하는 기존에 평가되지 않았던 신규한 일반 (기능성) 당류 소재를 대상으로 일련의 실험을 수행하였으며, 이 중 싸이코스가 에쿠올 생성을 촉진한다는 유의한 결과를 도출하고, 이로부터 본 발명을 완성하게 되었다.
본 발명은 싸이코스(Psicose)을 유효성분으로 하는 에쿠올 농도 상승제에 관한 것으로, 보다 구체적으로 인체 장내 미생물에 의한 에쿠올 생성을 촉진하기 위해 싸이코스를 유효성분으로 하는 에쿠올 농도 상승제에 관한 것이다.
본 발명의 주된 목적은 현재까지 보고된 일반 당류 및 다당류 외에 추가적으로 에쿠올의 전환을 촉진하는 효능을 가지는 기능성 식품을 제공하는 것이다.
본 발명의 다른 목적은 희소당류의 하나인 싸이코스(Psicose)의 새로운 용도를 제공하는 것이다.
본 발명의 또 다른 목적은 인체에 투여시 부작용없이 안전하고 생리 기능 활성화 작용을 가지며 에쿠올 전환을 촉진하는 기능을 갖는 조성물 혹은 식품을 제공하는 것이다.
본 발명은 상기 과제를 해결하기 위해 예의 노력을 거듭한 결과, 식품으로서 안전하고 저칼로리인 기능성 감미료 싸이코스(psicose)가 에쿠올 농도를 상승시키는 작용을 갖는 것을 발견하고 본 발명을 완성하게 되었다.
또한, 본 발명자들은 싸이코스를 함유한 배지를 사용하면 에쿠올 변환능을 갖는 미생물을 선택적으로 증식시키거나 에쿠올 변환능을 항진하는 기능을 갖는 것을 밝혀내었다.
본 발명은 인체에 투여시 부작용없이 안전하고 생리 기능 활성화 작용을 가지며 에쿠올 전환을 촉진하는 기능을 갖는 조성물 혹은 식품을 제공한다.
도 1은 다이드제인과 DHD로부터 에쿠올의 생성을 보여주는 전개 용 매(toluene:acetone = 2:1)을 사용한 TLC 사진이다(Dz:다이드제인, DHD:디하이드로다이드제인). 에쿠올 생성 미생물 군의 경우 3일까지 다이드제인을 사용하여 에쿠올을 생성하며, 3일째에는 모든 DHD가 에쿠올로 전환되었다.
도 2는 현재까지 알려진 장내미생물에 의한 다이드제인의 대사과정이다.
도 3은 에쿠올 변화에 미치는 각종 당류의 영향을 나타내는 그래프이다.
본 발명에 사용되는 싸이코스는 D-싸이코스를 의미하며, 당밀이나 이성화당 중에 미량 포함되어 있는데, D-프룩토오스로부터 에피메라제에 의해 효소적으로 생산가능한 희소당의 일종이다. 상기 싸이코스는 항산화성 등의 생리기능이 기대되고, 더욱이 감미가 설탕의 60~70%에 달하는 데도 불구하고 에너지가 거의 O 킬로칼로리이며, 용해성에 우수한 특성이 있다.
본 발명의 일 양태는 상기 싸이코스(Psicose)을 유효성분으로 하는 에쿠올 농도 상승제에 관한 것이다. 상기 싸이코스의 에쿠올 농도 상승 작용은 싸이코스가 에쿠올 변환능을 가진 미생물을 선택적으로 증식시키거나 미생물의 에쿠올 변환능을 항진시키는 것에 기인한다.
본 발명의 에쿠올 농도 상승 작용을 갖는 싸이코스는 체내, 혈중, 대장 내 등의 장내에서 에쿠올 농도상승제로 이용될 수 있으며, 이들 당류를 유효성분으로 하는 에쿠올 농도 상승제는 부정수소 등의 갱년기 장애, 골다공증, 고지혈증, 동맥경화, 유방암, 전립선암, 월경전증후군 등의 이소플라본이 관련된 각종 질병의 치료와 개선, 또는 그 예방 등의 목적에 이용될 수 있다.
통상적인 에쿠올 제조 방법으로서는, 다이드제인(daidzein)을 포함하는 원료에 대하여, 다이드제인을 대사하여 에쿠올을 생산하는 미생물(이하, 에쿠올 생산균이라 표기함)로 발효 처리하는 방법이 알려져 있다. 따라서, 본 발명의 싸이코스는 에쿠올의 기질인 다이드제인과 함께 사용하는 것이 바람직하다. 다이드제인은 합성품이나 천연추출물 등의 시판품을 사용해도 좋고, 다이드제인을 많이 함유하는 천연소재나 그의 가공품을 사용할 수 있다. 구체적으로 다이드제인을 많이 함유하는 소재로는 대두, 완두콩, 칡, 클로버 등을 들 수 있는데, 이들의 가공품으로는 두부, 두유, 튀김, 낫도, 간장, 된장, 템페이 등을 들 수 있다. 또한, 일반적으로 이소플라본 배당체는 체내의 장내세균의 작용에 의해 어글리콘화되기 때문에 다이드제인은 그 배당체 화합물인 다이드진, 말로닐다이드진, 아세틸다이드진 등의 형태로 사용될 수도 있다.
본원 발명에서 특별한 제한 없이 공지된 에쿠올 생산 균주를 이용할 수 있으며, 예를 들어, 박테로이데스 오바터스, 스트렙토코카스 인터메디어스, 스트펩토코카스 콘스테라터스, 락토코쿠스 가르비에 등이 있으며, 보다 상세히는 박테로이데스 E-23-15(FERM BP-6435호), 스트렙토코쿠스 E-23-17(FERM BP-6436호), 스트렙토코쿠스 A6G225(FERM BP-6437호), 락토코쿠스 20-92(FERM BP-10036호) 등이 있다.
본 발명은 상기한 통상적인 에쿠올 제조 방법에 있어서 싸이코스를 배지에 추가하여 에쿠올 생산균을 발효시킴으로써 에쿠올 생산 농도를 증가시킬 수 있다.
상기 양태에 있어서, 본 발명에 따른 에쿠올 농도 상승제는 싸이코스 외에 에쿠올 농도를 상승시키는 작용을 갖는 것으로 알려진 다른 당류를 추가로 포함할 수 있다. 추가될 수 있는 다른 당류는, 예를 들어, 아도니톨, 아라비노오즈, 에리스리톨, 갈락토오즈, 락티톨, 멜레지토오즈, 트레할로오즈, 리보오즈, 소르보오즈, 자일로스, 이노시톨 또는 솔비톨 등이 있으며, 이에 제한되는 것은 아니다.
따라서, 본 발명은 다른 양태는 에쿠올 생산 균주의 배양 배지에 싸이코스를 첨가하는 것을 포함하는 에쿠올 농도 상승 방법에 관한 것이다.
본 발명의 또 다른 양태는 싸이코스를 함유하는 것을 특징으로 하는 에쿠올 생성균주의 배양 배지를 제공하는 것이다.
본 발명의 또 다른 양태는 싸이코스(psicose)를 포함하는 에쿠올 농도 상승용 식품에 관한 것이다.
본 발명의 일 양태에서 다양한 장내 미생물의 집합체인 미생물 군을 사용하여 탄수화물의 종류에 따른 에쿠올 생성량의 차이를 확인하였다. 그 결과, 미생물 군총의 변화나 에쿠올 생성 미생물의 성장에 탄수화물이 영향을 미친다는 점을 확인하였다(실시예 2). 특히, 순수한 다이드제인를 사용하였을 때와는 달리 탄수화물을 첨가하였을 경우 에쿠올 생성량의 반응속도가 차이가 남을 보였다. 즉, 더욱 천천히 오랜 기간 동안 에쿠올의 생성이 이루어지는 것이다.
싸이코스가 3일 간의 반응조건에서 에쿠올 생성을 현저히 촉진하는 효과를 보였다 (실시예 2).
싸이코스의 에쿠올 농도 조절 활성은 에쿠올 생산능을 가진 미생물군 stock(에쿠올 프로듀서에서 채취한 분변을 원심 세정한 것, KR등록2008-0072635)에 에쿠올의 기질이 되는 다이드제인, 및 싸이코스를 첨가하여 배양하고, 배양액 중의 에쿠올 농도를 측정하여 다이드제인 및 자일로스를 함유하는 배양액 내의 에쿠올 농도와 비교하여 다음 식에 삽입하여 에쿠올 증감 백분율을 산출하였다.
에쿠올 증감 백분율(%) = (싸이코스 및 다이드제인을 함유하는 배양액 내의 에쿠올 농도)/ (자일로스 및 다이드제인을 함유하는 배양액 내의 에쿠올 농도) ×100
본 발명에 따른 에쿠올 농도 상승제의 투여량은 엄격한 제한은 없으며, 대상자나 적용 질환 등의 각종 사용 양태에 따라 얻어지는 효과가 다르기 때문에 적절한 투여량을 설정하는 것이 바람직하며, 그의 투여량은 1일당 0.1mg 내지 100g이며, 특히 50mg 내지 50g이 바람직하다.
본 발명의 에쿠올 농도 상승제는 경구 투여 또는 비경구투여 어느 것에도 사용할 수 있으나, 경구 투여가 바람직하다. 투여시에는 유효성분인 싸이코스를 경구투여, 직장내 투여, 주사 등의 투여방법에 적합한 고체 또는 액체의 의약용 무독성 담체와 혼합하여 관용의 의약품 제제형태로 투여할 수 있다.
이와 같은 제제로는 정제, 과립제, 산제, 캅셀제 등의 고형제, 용액제, 현탁제, 유제 등의 액제, 동결건조제 등을 들 수 있다. 이 제제들은 제제상의 통상의 방법으로 제조할 수 있다. 상기의 의약용 무독성 담체로는 전분, 덱스트린, 지방산글리세리드, 폴리에틸렌글리콜, 히드록시에틸렌전분, 에틸렌글리콜, 폴리옥시에틸렌소르비탄 지방산에스테르, 아미노산, 젤라틴, 알부민, 물, 생리식염수 등을 들 있다. 또, 필요에 따라 안정화제, 습윤제, 유화제, 결합제, 등장화제, 부형제 등의 일반 첨가제를 적절히 첨가할 수 있다.
또한, 본 발명의 당류는 상기와 같은 의약품 제제로서 사용할 수 있을 뿐 아니라, 음식품 등으로 사용할 수 있다. 이 경우에는 본 발명의 당류를 그대로 또는 각종 영양성분을 가하여 음식품 중에 함유시키면 좋다. 에쿠올 농도 상승작용을 갖는 싸이코스를 이용한 경우는 체내의 에쿠올 농도를 상승시킬 목적 또는 부정수소 등의 갱년기 장애, 골다공증, 고지혈증, 동맥경화, 유방암, 전립선암, 월경전증후군 등의 개선 및 예방 등에 유용한 보건용 식품 또는 식품소재로 이용할 수 있다.
구체적으로, 본 발명의 당류를 음식품에 배합하는 경우는, 음식품으로 사용할 수 있는 첨가제를 적의 사용하여 관용 수단을 사용하여 식용에 적합한 형태, 즉 과립상, 입상, 정제, 캅셀, 페이스트 등으로 성형해도 좋고, 또한 각종 식품, 예를 들면 햄, 소시지 등의 식육가공식품, 어묵, 오뎅 등의 수산가공식품, 빵, 과자, 버터, 분유, 발효유 제품에 첨가하여 사용하거나, 물, 과즙, 우유, 청량음료, 차음료 등의 음료에 첨가하여 사용해도 좋다. 또, 음식품에는 동물의 사료도 포함된다.
종래 알려진 에쿠올 생성 균주는 다이드제인의 환원 생성물인 디하이드로다이드제인(DHD)를 기질로만 사용하는 것으로 알려져 있으나, 미생물군 stock은 다양한 미생물이 있어 다이드제인에서 DHD의 반응이 진행되므로, 다이드제인과 DHD를 같이 기질로 사용하여 에쿠올을 생성하는 미생물 군을 사용하여 에쿠올의 생성을 TLC에서 분석하였으며, 탄수화물 존재 하에서의 에쿠올생성 변화량을 측정하였다.
실시예 1 : 에쿠올 생성 미생물군의 활성화와 표준 조건에서 에쿠올 생성 측 정
1) 에쿠올 생성 측정
액체 질소에 보관된 에쿠올 생성 미생물군 stock(에쿠올 프로듀서에서 채취한 분변을 원심 세정한 것, KR등록2008-0072635)을 GAM (Gifu anaerobic medium)
배지 (4ml)에 접종하여 3~4시간 동안 키워 활성화시켰다.
OD (optical density) 가 1.5정도에 이르렀을 때, 200㎕ 배지를 덜어 Eppendorf tube에 옮기고 최종 0.1 mM 농도의 다이드제인 농도로 첨가하고 vortexing하여 반응을 시작하였다.
시간에 따른 에쿠올 생성량을 측정하기 위해서 접종 후 각각 1일, 2일, 3일이 경과한 시점에 배지를 50㎕ 씩 덜어내었다.
에틸 아세테이트 1ml를 배지에 첨가하여 이소플라노이드를 추출하였다. 추출과정은 vortexing 후 원심분리하여 에틸아세테이트 800㎕를 취하고 vaccum dry로 용매를 제거한 후 메탄올 10㎕에 녹인 후 2㎕를 TLC에 점적하였다.
1) 결과
상기 TLC에서 미생물 군이 다이드제인으로부터 에쿠올을 생성하는 것으로 확인되었으며, 그 생성량은 접종 후 3일간 계속 증가하였다. 3일 후에는 기질로 공급된 다이드제인이 검출되지 않았다. 따라서, 미생물 전환 실험은 3일간 진행되었다.
실시예 2 : 탄수화물 존재 하에서의 에쿠올 생성 변화량 측정
자일로스, 타가토스, 싸이코스, 수크로스, 프락토올리고당, 글루코스등 총 6종의 탄수화물을 사용하여 실시예 1에 이용된 에쿠올 생성 조건에서 에쿠올 생성 증감량을 측정하였다.
1) 미생물 반응 및 에쿠올 생성량 비교
상기 실시예 1과 동일한 방법으로 에쿠올 생성 미생물 군을 접종하고 생성물을 추출하였으며, 차이점은 활성화된 미생물 군의 기질 접종 전에 자일로스, 타가토스, 싸이코스, 수크로스, 프락토올리고당, 글루코스를 각각 2mg씩 배지에 첨가하여 최종 탄수화물 농도를 1%로 조정하는 과정을 추가한 것이다. 또한, 기질로는 다이드제인만을 사용하였다. 순수한 다이드제인을 사용하였을 때와는 달리 탄수화물을 첨가하였을 경우 에쿠올 생성량의 반응속도에 있어 차이가 있었다. 즉, 더욱 천천히 오랜 기간동안 에쿠올의 생성이 이루어 지는 것이다.
생성물 중 에쿠올 증감량을 비교하기 위해서 TLC 결과의 image analyer를 사용한 정량 분석을 사용하였다.
2) 결과
실험결과 자일로스 및 다이드제인을 함유하는 배양액 내의 에쿠올 생성량을 100으로 하여 3일째 각 탄수화물 첨가시의 에쿠올 생성량을 비교하였을 때, 비교 결과는 아래와 같다(아래 표 2의 결과는 각 탄수화물 종류에 따른 에쿠올의 생성량을 자일로스와 상대비교(%)한 것으로 2가지 발색 방법에 대한 이미지 분석결과의 평균값을 나타낸 것이다).
표 2
탄수화물 | 자일로스 | 타가토스 | 싸이코스 | 수크로스 | 프락토올리고당 | 글루코스 |
증감량(%) | 100 | 54 | 139 | 54 | 55 | 24 |
실시예 3 : 정제 형태의 에쿠올 농도 상승제
하기의 처방으로 각종 성분을 혼합하여 조립, 건조 후에 타정하여 정제를 제조하였다.
미결정 세룰로오스 110mg
싸이코스 75mg
스테아린산 마그네슘 0.4mg
메틸세룰로오스 13mg
실시예 4 : 청량음료 형태의 에쿠올 농도 상승제
하기의 처방으로 각종 성분을 혼합하고 균질화하여 청량음료를 수득하였다.
향료 0.7 g
싸이코스 2.0 g
시트르산 0.4 g
과당 5 g
정제수 80 g
Claims (5)
- 싸이코스(psicose)을 유효성분으로 포함하는 에쿠올 농도 상승제.
- 제1항에 있어서,상기 싸이코스가 에쿠올 변환능을 가진 미생물을 선택적으로 증식시키거나 미생물의 에쿠올 변환능을 항진하는 기능을 갖는 에쿠올 농도 상승제.
- 에쿠올 생산 균주의 배양 배지에 싸이코스를 첨가하는 것을 포함하는 에쿠올 의 농도를 상승시키는 방법.
- 제3항에 있어서,상기 사이코스가 에쿠올 변환능을 가진 미생물을 선택적으로 증식시키거나 미생물의 에쿠올 변환능을 항진하는 기능을 갖는, 방법.
- 싸이코스(psicose)을 유효성분으로 포함하는 에쿠올 농도 상승용 식품.
Priority Applications (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP12852912.0A EP2787071B1 (en) | 2011-11-30 | 2012-10-15 | Equol concentration increasing agent containing psicose as active ingredient |
JP2014544650A JP5936704B2 (ja) | 2011-11-30 | 2012-10-15 | サイコースを有効成分とするエクオール濃度上昇剤 |
EP16205334.2A EP3170502B1 (en) | 2011-11-30 | 2012-10-15 | Combination of psicose and daidzein as a medicinal agent |
US14/361,528 US9504703B2 (en) | 2011-11-30 | 2012-10-15 | Equol concentration increasing agent containing psicose as active ingredient |
US15/209,266 US9839647B2 (en) | 2011-11-30 | 2016-07-13 | Equol concentration increasing agent containing psicose as active ingredient |
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KR1020110126552A KR101307947B1 (ko) | 2011-11-30 | 2011-11-30 | 싸이코스를 유효성분으로 하는 에쿠올 농도 상승제 |
KR10-2011-0126552 | 2011-11-30 |
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US14/361,528 A-371-Of-International US9504703B2 (en) | 2011-11-30 | 2012-10-15 | Equol concentration increasing agent containing psicose as active ingredient |
US15/209,266 Continuation US9839647B2 (en) | 2011-11-30 | 2016-07-13 | Equol concentration increasing agent containing psicose as active ingredient |
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WO2013081294A1 true WO2013081294A1 (ko) | 2013-06-06 |
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PCT/KR2012/008398 WO2013081294A1 (ko) | 2011-11-30 | 2012-10-15 | 싸이코스를 유효성분으로 하는 에쿠올 농도 상승제 |
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US (2) | US9504703B2 (ko) |
EP (2) | EP2787071B1 (ko) |
JP (1) | JP5936704B2 (ko) |
KR (1) | KR101307947B1 (ko) |
WO (1) | WO2013081294A1 (ko) |
Cited By (3)
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US9259022B2 (en) | 2013-03-15 | 2016-02-16 | Tate & Lyle Ingredients Americas Llc | Sweetener |
US9491960B2 (en) | 2013-03-15 | 2016-11-15 | Tate & Lyle Ingredients Americas Llc | Sweetener |
US9854827B2 (en) | 2013-03-15 | 2018-01-02 | Tate & Lyle Ingredients Americas Llc | Sweetener |
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SG11202013008UA (en) * | 2018-07-10 | 2021-02-25 | Univ Keio | Intestinal microbe having d-psicose-responsive proliferation |
KR102079003B1 (ko) | 2018-08-10 | 2020-02-19 | 서울대학교산학협력단 | 활성이 증대된 테트라하이드로다이드제인 환원효소 및 에쿠올 유도체 생산에의 응용 |
JP6800386B1 (ja) * | 2020-01-23 | 2020-12-16 | 三栄源エフ・エフ・アイ株式会社 | 骨代謝改善用組成物 |
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Publication number | Priority date | Publication date | Assignee | Title |
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US9259022B2 (en) | 2013-03-15 | 2016-02-16 | Tate & Lyle Ingredients Americas Llc | Sweetener |
US9491960B2 (en) | 2013-03-15 | 2016-11-15 | Tate & Lyle Ingredients Americas Llc | Sweetener |
US9635879B2 (en) | 2013-03-15 | 2017-05-02 | Tate & Lyle Ingredients Americas Llc | Sweetener |
US9854827B2 (en) | 2013-03-15 | 2018-01-02 | Tate & Lyle Ingredients Americas Llc | Sweetener |
Also Published As
Publication number | Publication date |
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KR20130060477A (ko) | 2013-06-10 |
US20160361331A1 (en) | 2016-12-15 |
JP2015500802A (ja) | 2015-01-08 |
EP3170502B1 (en) | 2019-04-24 |
JP5936704B2 (ja) | 2016-06-22 |
US9504703B2 (en) | 2016-11-29 |
EP2787071B1 (en) | 2018-11-14 |
EP2787071A1 (en) | 2014-10-08 |
US20140329286A1 (en) | 2014-11-06 |
US9839647B2 (en) | 2017-12-12 |
EP2787071A4 (en) | 2015-06-24 |
EP3170502A1 (en) | 2017-05-24 |
KR101307947B1 (ko) | 2013-09-12 |
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