JP2013252125A - Method for producing liquefied koji with enhanced antioxidative activity - Google Patents
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本発明は、抗酸化活性が増強した液化麹およびその製造方法、該液化麹を含む抗酸化剤、ならびに機能性食品などの各種組成物に関する。本発明はまた、麹または液化麹を有効成分として含む認知症の治療及び/又は予防剤に関する。 The present invention relates to a liquefied koji with enhanced antioxidant activity and a method for producing the same, an antioxidant containing the liquefied koji, and various compositions such as functional foods. The present invention also relates to a therapeutic and / or prophylactic agent for dementia comprising sputum or liquefied sputum as an active ingredient.
日本の伝統的な清酒、焼酎、味噌、醤油などの発酵食品には、抗酸化作用をはじめとした多くの機能性があることが近年明らかにされている。その機能性成分の多くは「麹」由来の酵素反応や、それに続く加熱や貯蔵中におこるメイラード反応により生成することが報告されている(非特許文献1、2等)。
In recent years, fermented foods such as traditional Japanese sake, shochu, miso, and soy sauce have been revealed to have many functions including antioxidant activity. It has been reported that many of the functional components are produced by an enzyme reaction derived from “mochi” and subsequent Maillard reaction that occurs during heating and storage (Non-patent
また、麹は糖化酵素をはじめとする酵素の生産により発酵の安全に寄与し、香味の付与に大きく貢献している。そのため、麹づくり(製麹)はそれぞれの発酵食品の最終製品に適した方法により行われている。たとえば清酒においては麹の温度を徐々に高め最終的に約40℃にすることにより清酒醸造に適した酵素の生産を促すのに対し、本格焼酎においては約37℃で麹が増殖した段階で約35℃に温度を下げ、クエン酸の生成を促すなどの違いが見られる。このように、製麹段階では、原料を糖化・液化するための酵素生成や、もろみを腐敗から守るクエン酸生成を行なうことに主眼が置かれており、抗酸化性などの機能性を高めることは意図されていない。また、製麹期間は、麹菌体の過剰な増殖は生産性の低下、風味への悪影響をもたらすことから通常は2日程度である。 In addition, koji contributes to the safety of fermentation by producing enzymes such as saccharifying enzymes, and greatly contributes to imparting flavor. Therefore, the making of koji (making of koji) is performed by a method suitable for the final product of each fermented food. For example, in sake, the temperature of koji is gradually increased to about 40 ° C to promote the production of enzymes suitable for sake brewing, while in the case of full-fledged shochu, the koji grows at about 37 ° C. Differences are seen such as lowering the temperature to 35 ° C and promoting the production of citric acid. Thus, in the koji making stage, the main focus is on the production of enzymes for saccharification and liquefaction of raw materials, and the production of citric acid that protects moromi from spoilage. Is not intended. In addition, the koji-making period is usually about 2 days because excessive growth of koji fungi causes a decrease in productivity and an adverse effect on flavor.
製麹で得られた麹は、その後、発酵食品の種類によって用い方は異なるが、一般的には、水および酵母と混合し、発酵させてもろみとし、これに主原料(蒸米、麦、甘藷など)を仕込んで発酵、熟成させる。従って、製麹に続いて麹の機能性を高める加工処理を施すことは通常の発酵食品の製造においては行なわれていない。 The koji obtained from koji making is then used differently depending on the type of fermented food, but in general, it is mixed with water and yeast and fermented into moromi, and the main ingredients (steamed rice, wheat, sweet potato) Etc.) is fermented and matured. Therefore, the process of improving the functionality of koji following koji making is not performed in the production of ordinary fermented foods.
これまで機能性飲食品や医薬品の製造過程で麹に加熱が行われている例はある。例えば、特許文献1は紫芋に米麹を加えて50〜70℃で18〜28時間発酵させた紫芋発酵液を主成分とする脳保護作用剤を開示するが、その機能性は紫芋に由来するものであり、米麹は甘みの増強に寄与しているのみである。また、特許文献2は米粉砕物に麹を加え55℃16時間放置した後、固液分離により得たろ液を85℃で30分加熱したものを有効成分とするがん細胞休眠剤を開示するが、麹は糖化のために使用され、後半の高温加熱は殺菌を目的としたものである。よって、これらの例はいずれも加熱処理は、製麹後の麹を他の澱粉材料に混合してから行われており、製麹の麹をそのまま加熱処理に供したものではない。一方、特許文献3には、黒米由来のアントシアニンを含む組成物の製造方法が開示されており、その製造工程において製麹後の麹に水を加えて糖化液を製造する工程が含まれるが、糖化温度と時間は50〜60℃、10〜20時間が好ましいとされており、その後に酵素失活と殺菌のために80〜100℃の高温加熱が行なわれているが、時間は10分程度にすぎない。
There has been an example in which heating has been performed in the process of manufacturing functional foods and beverages and pharmaceuticals. For example,
従って、麹を用いる発酵食品には抗酸化活性をはじめとする多くの機能性があることは知られているが、麹そのものを機能性食品素材として捉えた研究は少なく、また、これまで加熱処理と麹の抗酸化作用の増強との関係については報告するものはない。 Therefore, it is known that fermented foods using koji have many functionalities such as antioxidant activity, but there are few studies on koji itself as a functional food material. There is no report on the relationship between the anti-oxidant effect of sorghum and sputum.
一方、認知症は、年齢が上がるにつれて発症率が高くなると言われており、今後、ますます高齢化社会が進むにつれて、認知症への対応が大きな課題となっている。認知症への取り組みとしては、まず早期に発見し、適切な診断・治療を行なうことによって症状の進行を遅らせること、認知症発症の予防に努めることが大切である。認知症は、加齢や脳の疾患など種々の原因により、脳の細胞が死んでしまい、学習・記憶能力や判断力などの脳機能が低下するために様々な障害がおこり、日常生活に支障をきたす状態をいう。認知症の中でもアルツハイマー型は過半数を占め、レビー小体型や脳血管型がこれに次ぐ。アルツハイマー型認知症は、新しい記憶が一時保存される「海馬」が損傷するために新しい物事を忘れるという症状が現れる。また、脳神経細胞中のミトコンドリアで発生する活性酸素や過酸化脂質による酸化ストレスは、年齢とともに増加し、神経細胞損傷や老化を加速化する。酸化ストレスはアルツハイマー型認知症の原因となることがわかっており、脳の酸化ストレスを軽減できる優れた抗酸化物質が発見できれば、認知機能低下を抑制でき、認知症を改善できることが期待される。これまで、アルツハイマー型認知症の治療薬としては、代表的にはアセチルコリンエステラーゼ阻害薬(塩酸ドネペシルなど)のほか、非ステロイド系抗炎症薬、β-セクレターゼ阻害薬などがある。また、日常的に摂取してアルツハイマー型認知症を防ぎ、進行を遅らせる効果が報告される野菜や果物等の天然由来成分としては、ウコンに含まれるクルクミン、葡萄に含まれるレスベラトール、米ぬかに含まれるフェルラ酸、イチョウの葉エキスなどがあるが、麹については知られていない。 On the other hand, it is said that the incidence of dementia increases as the age increases, and as the aging society progresses in the future, dealing with dementia has become a major issue. As an approach to dementia, it is important to first detect it early, to delay the progression of symptoms by appropriate diagnosis and treatment, and to try to prevent the onset of dementia. Dementia is caused by various causes such as aging and brain diseases, and the brain cells die and brain functions such as learning / memory ability and judgment decline. The state that causes Alzheimer type accounts for the majority of dementia, followed by Lewy body type and cerebrovascular type. Alzheimer-type dementia has the symptom of forgetting new things because the “hippocampus” where new memories are temporarily stored is damaged. In addition, oxidative stress caused by active oxygen and lipid peroxide generated in mitochondria in brain neurons increases with age, accelerating nerve cell damage and aging. Oxidative stress is known to cause Alzheimer-type dementia. If an excellent antioxidant capable of reducing brain oxidative stress can be found, it is expected that cognitive decline can be suppressed and dementia can be improved. To date, typical treatments for Alzheimer-type dementia include acetylcholinesterase inhibitors (such as donepecil hydrochloride), non-steroidal anti-inflammatory drugs, and β-secretase inhibitors. In addition, naturally-occurring ingredients such as vegetables and fruits that are reported to have an effect of preventing Alzheimer-type dementia and delaying progression by daily consumption include curcumin contained in turmeric, resveratrol contained in rice bran, and rice bran There are ferulic acid, ginkgo biloba extract, etc., but cocoon is not known.
本発明は、発酵食品の機能性成分である麹の抗酸化活性を高め、麹自体を機能性食品などの素材として利用すること、および麹の新たな機能を見出すことを課題とする。 An object of the present invention is to increase the antioxidant activity of koji, which is a functional component of fermented food, to use koji itself as a material for functional food, and to find a new function of koji.
本発明者らは、上記課題を解決すべく鋭意研究を重ねた結果、麹を特定の条件で加熱処理するという簡潔な手法で、麹中のスーパーオキシドアニオンラジカル消去能 (SOSA)及び酸素ラジカル吸収能(ORAC)が飛躍的に上昇するとともに、抗酸化物質である5−ヒドロキシメチルフルフラール(HMF)含量が増加することを確認し、抗酸化活性が増強されるという知見を得た。 As a result of intensive studies to solve the above-mentioned problems, the present inventors have conducted a simple method of heat-treating soot under specific conditions, so that superoxide anion radical scavenging ability (SOSA) and oxygen radical absorption in soot It was confirmed that the activity (ORAC) increased dramatically and the content of 5-hydroxymethylfurfural (HMF), which is an antioxidant, increased, and the antioxidant activity was enhanced.
本発明者らはまた、麹の新たな機能性の発見と評価を目的として、老化促進マウスに米麹または液化麹を摂取させたところ、当該マウスの学習・記憶能力が亢進し、脳のアミロイドβ蓄積が減少することを確認し、麹に認知症改善効果があることを見出した。
本発明はかかる知見をもとに完成されたものである。
For the purpose of discovering and evaluating the new functionality of sputum, the present inventors also ingested rice bran or liquefied sputum into an aging-promoted mouse, which increased the learning / memory ability of the mouse, and increased amyloid in the brain. It was confirmed that β accumulation decreased, and it was found that wrinkles have a dementia improving effect.
The present invention has been completed based on such knowledge.
すなわち、本発明は以下の発明を包含する。
(1) 麹を50℃〜60℃の温度で6〜8日間加熱処理する工程を含む、抗酸化活性が増強した液化麹の製造方法。
(2) 麹を50℃〜60℃の温度で1〜3日間加熱処理する工程、続いて70℃〜80℃の温度で4〜6日間加熱処理する工程を含む、抗酸化活性が増強した液化麹の製造方法。
(3) 50℃〜60℃の加熱処理を2日間行い、70℃〜80℃の加熱処理を5日間行う、(2)に記載の製造方法。
(4) 前記麹が、乾燥麹と水の当量混合物である、(1)〜(3)のいずれかに記載の製造方法。
(5) 前記麹が、白麹、黄麹、および黒麹からなる群より選択される、(1)〜(4)のいずれかに記載の製造方法。
(6) (1)〜(5)のいずれかに記載の製造方法によって得られる抗酸化活性が増強した液化麹。
That is, the present invention includes the following inventions.
(1) A method for producing a liquefied koji with enhanced antioxidant activity, comprising a step of heat-treating koji at a temperature of 50 ° C. to 60 ° C. for 6 to 8 days.
(2) Liquefaction with enhanced antioxidative activity, including a step of heat-treating koji at a temperature of 50 ° C. to 60 ° C. for 1 to 3 days, followed by a heat treatment of 70 to 80 ° C. for 4 to 6 days A method for producing firewood.
(3) The production method according to (2), wherein the heat treatment at 50 ° C. to 60 ° C. is performed for 2 days, and the heat treatment at 70 ° C. to 80 ° C. is performed for 5 days.
(4) The production method according to any one of (1) to (3), wherein the soot is an equivalent mixture of dry soot and water.
(5) The production method according to any one of (1) to (4), wherein the wrinkle is selected from the group consisting of white glaze, jaundice, and black glaze.
(6) A liquefied koji with enhanced antioxidant activity obtained by the production method according to any one of (1) to (5).
(7) (6)に記載の液化麹を含む、抗酸化剤。
(8) (7)に記載の抗酸化剤を含む、飲食品。
(9) 前記飲食品が、健康食品、機能性食品、栄養補助食品、または特定保健用食品である、(8)に記載の飲食品。
(10) (7)に記載の抗酸化剤を含む、医薬品、医薬部外品、または化粧品。
(11) 麹または液化麹を有効成分として含有する認知症の治療及び/又は予防剤。
(12) 液化麹が、(1)〜(5)のいずれかに記載の製造方法によって得られる液化麹である、(11)に記載の認知症の治療及び/又は予防剤。
(13) 認知症が、酸化ストレスに起因する認知症である、(11)または(12)に記載の認知症の治療及び/又は予防剤。
(7) An antioxidant comprising the liquefied soot according to (6).
(8) A food or drink comprising the antioxidant according to (7).
(9) The food or drink according to (8), wherein the food or drink is a health food, a functional food, a nutritional supplement, or a food for specified health use.
(10) A pharmaceutical, quasi-drug or cosmetic comprising the antioxidant according to (7).
(11) A therapeutic and / or prophylactic agent for dementia containing sputum or liquefied sputum as an active ingredient.
(12) The therapeutic and / or preventive agent for dementia according to (11), wherein the liquefied sputum is a liquefied sputum obtained by the production method according to any one of (1) to (5).
(13) The therapeutic and / or preventive agent for dementia according to (11) or (12), wherein the dementia is dementia caused by oxidative stress.
本発明の方法によれば、加熱処理という簡便な操作のみで麹の抗酸化活性を飛躍的に向上させることできる。本発明により得られる抗酸化活性が増強した液化麹は、抗酸化剤として機能性食品や医薬品等の機能性素材として有用である。また、当該液化麹は、学習・記憶能力を亢進し、脳のアミロイドβ蓄積を減少させることができるため、認知症の改善及び/又は予防に有効である。 According to the method of the present invention, it is possible to dramatically improve the antioxidant activity of koji only by a simple operation of heat treatment. The liquefied koji with enhanced antioxidant activity obtained by the present invention is useful as a functional material for functional foods and pharmaceuticals as an antioxidant. In addition, the liquefied sputum is effective in improving and / or preventing dementia because it can enhance learning and memory ability and reduce amyloid β accumulation in the brain.
以下に、本発明について詳細に述べる。
本発明の抗酸化活性が増強した液化麹の製造方法は、麹の加熱処理を一定の温度条件下で行なう。
The present invention will be described in detail below.
In the method for producing a liquefied soot with enhanced antioxidant activity according to the present invention, heat treatment of the soot is performed under a constant temperature condition.
第1の方法は、麹の加熱を一段階で行なう方法(一段階法)で、この方法では麹を50℃〜60℃で6〜8日間加熱処理する。 The first method is a method in which the soot is heated in one step (one-step method). In this method, the soot is heated at 50 to 60 ° C. for 6 to 8 days.
第2の方法は、麹の加熱を二段階で行なう方法(二段階法)で、この方法では麹を50℃〜60℃の温度で1〜3日間加熱処理し、続いて70℃〜80℃の温度で4〜6日間加熱処理する。 The second method is a method in which the soot is heated in two stages (two-stage method). In this method, the soot is heated at a temperature of 50 ° C. to 60 ° C. for 1 to 3 days, followed by 70 ° C. to 80 ° C. For 4 to 6 days.
一段階法または二段階法のいずれによっても未加熱麹よりも優れた抗酸化作用を有する麹が得られるが、抗酸化作用がより増強した麹が得られる上で二段階法が好ましい。 Either a one-step method or a two-step method can provide a soot having an antioxidant effect superior to that of unheated soot, but a two-step method is preferred in terms of obtaining a soot with an enhanced antioxidant effect.
加熱処理の温度および時間は上記の範囲で適宜変更することができる。一段階法および二段階法の前段の加熱温度は50℃〜60℃であればよく、54〜56℃が好ましく、55℃がより好ましい。また、二段階法の後段の加熱温度は70℃〜80℃であればよく、74〜76℃が好ましく、75℃がより好ましい。 The temperature and time of the heat treatment can be appropriately changed within the above range. The heating temperature in the previous stage of the one-step method and the two-step method may be 50 ° C to 60 ° C, preferably 54 to 56 ° C, and more preferably 55 ° C. Moreover, the heating temperature of the latter stage of a two-stage method should just be 70 to 80 degreeC, 74 to 76 degreeC is preferable and 75 degreeC is more preferable.
本発明方法の最も好ましい加熱条件は、55℃で2日間、続いて75℃で5日間行うことである。 The most preferred heating condition of the method of the present invention is to carry out at 55 ° C. for 2 days, followed by 75 ° C. for 5 days.
加熱処理は、断熱密閉容器等の温度調節機能のついた当分野で常套的に用いられる穀類の液化装置を用いて行なえばよい。 The heat treatment may be performed using a cereal liquefaction apparatus conventionally used in the art having a temperature control function, such as a heat-insulated airtight container.
本発明に用いる麹の原料は、米、麦、芋、豆類等のいずれでもよいが、酵素生産性や製麹の容易性の観点から、米が好ましい。 The raw material of koji used in the present invention may be any of rice, wheat, koji, beans, etc., but rice is preferred from the viewpoint of enzyme productivity and ease of koji making.
また、麹の種類としては、製麹の際に上記原料に添加する麹菌の種類により黄麹、黒麹、白麹等があり、これらのいずれもよいが、クエン酸の生成やHMF生産性の点から白麹が望ましい。 The types of koji include yellow koji, black koji, and white koji depending on the type of koji mold added to the raw material during koji making. Any of these may be used, but the production of citric acid and HMF productivity From the point, white birch is desirable.
麹の調製(製麹)は、原料に応じて通常の方法で行なえばよい。例えば、米麹の場合、
蒸米を浸漬後蒸煮し、冷却後、白麹菌(Aspergillus kawachii)、黄麹菌(Aspergillus oryzae)、黒麹菌(Aspergillus awamori)などの麹菌の胞子(種麹)を接菌し、37℃程度に維持し、仲仕事を経て仕舞い仕事のあと35℃程度に維持し、クエン酸の生成を図り、種掛け後約40時間後に出麹とする。
The preparation of the koji (making of koji) may be performed by a normal method depending on the raw material. For example, in the case of rice bran,
Steamed rice is soaked and boiled, cooled, and then inoculated with Aspergillus spores (seeds) such as Aspergillus kawachii, Aspergillus oryzae, Aspergillus awamori, and maintained at about 37 ° C. After the middle work, the temperature is maintained at about 35 ° C. after completion of the work, and citric acid is produced.
上記の製麹で得られた麹はそのまま加熱処理に供することもできるが、長期保存するときは凍結乾燥等で、乾燥麹とする。乾燥麹を用いる場合は、加熱処理は、乾燥麹に水、好ましくは55℃程度の湯を加えた混合物に対して行なう。乾燥麹と水との混合割合は等量であることが好ましい。乾燥麹に対して水が多すぎると成分が希釈され所望の抗酸化活性が得られず、後で濃縮工程が必要になる場合がある。また、水が過剰であると微生物汚染により腐敗する恐れがある。一方、乾燥麹に対して水が少なすぎると液化が進行しづらいために機能性成分の生成が少なくなってしまう。 The koji obtained by the above koji making can be subjected to a heat treatment as it is, but when stored for a long time, it is made into a dried koji by freeze drying or the like. When using a dry cake, the heat treatment is performed on a mixture obtained by adding water, preferably about 55 ° C. hot water, to the dried cake. The mixing ratio of the dried soot and water is preferably equal. If there is too much water for the dried soot, the components may be diluted and the desired antioxidant activity may not be obtained, and a concentration step may be required later. Moreover, if water is excessive, there is a risk of rot due to microbial contamination. On the other hand, if the amount of water is too small relative to the dried soot, liquefaction is difficult to proceed and the generation of functional components is reduced.
上記の麹の加熱処理により得られる液化麹は、未処理の麹に比べてスーパーオキシドラジカル消去能(SOSA)及び酸素ラジカル吸収能(ORAC)が有意に増加し、優れた抗酸化作用を有する。したがって、上記の液化麹は、抗酸化剤として使用できる。 The liquefied soot obtained by the above heat treatment of soot has a significantly increased superoxide radical scavenging ability (SOSA) and oxygen radical absorption ability (ORAC) compared to untreated soot, and has an excellent antioxidant action. Therefore, the above liquefied soot can be used as an antioxidant.
本発明の抗酸化剤は、そのまま用いることも可能であるが、本発明の効果を損なわない範囲で適宜他の成分を配合し、飲食品、医薬品、医薬部外品、化粧品等の各種組成物として提供することが好ましい。 The antioxidant of the present invention can be used as it is, but various other components such as foods and drinks, pharmaceuticals, quasi-drugs, cosmetics, etc. are appropriately blended within a range not impairing the effects of the present invention. It is preferable to provide as.
本発明において、飲食品とは、ヒトが摂取するための飲食品をいい、健康食品、機能性食品、栄養補助食品、または特定保健用食品を含む意味で用いられる。 In the present invention, the food / beverage product refers to a food / beverage product to be ingested by humans, and is used to include a health food, a functional food, a nutritional supplement, or a food for specified health use.
飲食品の形態は、食用に適した形態、例えば、固形状、液状、顆粒状、粒状、粉末状、カプセル状、クリーム状、ペースト状のいずれであってもよい。 The form of the food or drink may be any form suitable for edible use, for example, solid, liquid, granular, granular, powder, capsule, cream, or paste.
飲食品の種類としては、具体的には、食パン、菓子パン等のパン類;そば、うどん、中華麺等の麺類;菓子類;加工乳、発酵乳、ヨーグルト、バター、チーズ等の乳製品;かまぼこ、ちくわ、ハム、ソーセージ等の水産・畜産加工食品;マーガリン、マヨネーズ、ショートニング、ホイップクリーム、ドレッシング等の油脂及び油脂加工食品;しょうゆ、ソース、焼肉のたれ、酢、みりん等の調味料;清涼飲料、炭酸飲料、栄養飲料、果実飲料、乳飲料など飲料(これらの飲料の濃縮原液及び調整用粉末を含む)が挙げられるが、これらに限定はされない。 Specific types of foods and drinks include breads such as bread and confectionery bread; noodles such as buckwheat, udon and Chinese noodles; confectionery; dairy products such as processed milk, fermented milk, yogurt, butter and cheese; kamaboko Processed fishery and livestock products such as chikuwa, ham and sausage; fats and oils processed foods such as margarine, mayonnaise, shortening, whipped cream and dressing; seasonings such as soy sauce, sauce, grilled meat sauce, vinegar, mirin; soft drinks , Beverages such as carbonated beverages, nutritional beverages, fruit beverages and milk beverages (including concentrated concentrates of these beverages and powders for adjustment), but are not limited thereto.
また、本発明の飲食品には麹を本来利用する発酵食品も含まれることはいうまでもない。
本発明の製造方法で得られた液化麹を用いて清酒を製造する場合は、酒母やもろみの仕込み段階において、焼酎を製造する場合にはもろみの仕込み段階において、醤油を製造する場合は、盛り込みの段階において、味噌を製造する場合には、仕込みの段階において、通常の麹に代えて用いることができる。
In addition, it goes without saying that the food and drink of the present invention includes fermented foods that originally use koji.
In the case of producing sake using the liquefied koji obtained by the production method of the present invention, in the step of preparing the sake mother or moromi, in the case of producing shochu, in the step of preparing the moromi, in the case of producing soy sauce, In the case of producing miso at this stage, it can be used in place of ordinary koji at the stage of preparation.
本発明の飲食品は、その種類に応じて通常使用される添加物を適宜配合してもよい。添加物としては、食品衛生上許容されうる添加物であればいずれも使用できるが、例えば、砂糖、果糖、異性化液糖、ブドウ糖、アスパルテーム、ステビア等の甘味料;クエン酸、リンゴ酸、酒石酸等の酸味料;デキストリン、澱粉等の賦形剤;結合剤、希釈剤、香料、着色料、緩衝剤、増粘剤、ゲル化剤、安定剤、保存剤、乳化剤、分散剤、懸濁化剤、防腐剤などが挙げられる。 The food / beverage products of the present invention may be appropriately blended with additives usually used depending on the type. Any additive that is acceptable for food hygiene can be used as the additive. For example, sweeteners such as sugar, fructose, isomerized liquid sugar, glucose, aspartame, stevia; citric acid, malic acid, tartaric acid Acidulants such as dextrin, starch, etc .; binders, diluents, fragrances, colorants, buffers, thickeners, gelling agents, stabilizers, preservatives, emulsifiers, dispersants, suspensions Agents, preservatives and the like.
本発明の飲食品における液化麹の配合量は、その抗酸化作用が発揮できる量であればよいが、対象飲食品の一般的な摂取量、飲食品の形態、効能・効果、呈味性、嗜好性およびコストなどを考慮して適宜設定すればよい。 The blending amount of the liquefied koji in the food and drink of the present invention may be an amount that can exhibit its antioxidant action, but the general intake of the target food and drink, the form of the food and drink, the efficacy and effect, the taste, What is necessary is just to set suitably in consideration of palatability and cost.
また、本発明の飲食品は、顕著な抗酸化作用、すなわち、活性酸素消去作用を有するので、生体内に存在する活性酸素に起因して生じる各種の疾患や障害の予防または改善にも有効である。活性酸素に起因して生じる各種の疾患や障害としては、例えば、動脈硬化や心筋梗塞などの循環器系疾患、胃・十二指腸潰瘍・胃粘膜障害、潰瘍性大腸炎、過敏性大腸炎などの消化器官系疾患、薬物や有害物質による肝障害、虚血性再灌流障害、糖尿病の合併症、白内障や加齢黄斑変性などの眼科疾患、シミ・シワなどの皮膚の変性、歯周病などの炎症性疾患、癌、老化などが挙げられる。 In addition, since the food and drink of the present invention has a remarkable antioxidant action, that is, an active oxygen scavenging action, it is effective for preventing or improving various diseases and disorders caused by active oxygen present in the living body. is there. Various diseases and disorders caused by active oxygen include, for example, cardiovascular diseases such as arteriosclerosis and myocardial infarction, digestion such as gastric / duodenal ulcer / gastric mucosal disorder, ulcerative colitis, irritable colitis, etc. Organ system diseases, liver damage due to drugs and harmful substances, ischemic reperfusion injury, diabetic complications, ophthalmic diseases such as cataracts and age-related macular degeneration, skin degeneration such as spots and wrinkles, inflammation such as periodontal disease Disease, cancer, aging, etc. are mentioned.
また、本発明の抗酸化剤を医薬品として提供する場合は、医薬上許容され、かつ剤型に応じて適宜選択した適当な添加剤(例えば担体、賦形剤、希釈剤、結合剤、滑沢剤、崩壊剤又は崩壊補助剤、可溶化剤、安定化剤、保存剤、防腐剤、増量剤、分散剤等)を用いて、公知の種々の方法にて経口又は非経口的に全身又は局所投与することができる各種製剤形態に調製すればよい。 In addition, when the antioxidant of the present invention is provided as a pharmaceutical, an appropriate additive (for example, carrier, excipient, diluent, binder, lubricant) that is pharmaceutically acceptable and appropriately selected depending on the dosage form. Agents, disintegrants or disintegration aids, solubilizers, stabilizers, preservatives, preservatives, extenders, dispersants, etc.) in various known ways, orally or parenterally systemically or locally. What is necessary is just to prepare in the various formulation form which can be administered.
本発明の医薬品は、経口または非経口的に投与することができるが、好ましくは経口投与である。本発明の医薬品を経口投与する場合は、錠剤(糖衣錠を含む)、カプセル剤、顆粒剤、散剤、丸剤、内用水剤、懸濁液剤、乳剤、シロップ剤等に製剤化するか、使用する際に再溶解させる乾燥生成物にしてもよい。また、本発明の医薬品を非経口投与する場合は、注射剤(例えば、皮下注射剤、静脈内注射剤、筋肉内注射剤、腹腔内注射剤)、点滴剤、坐剤などに製剤化し、注射用製剤の場合は単位投与量アンプル又は多投与量容器の状態で提供される。製剤化に当たっては、本発明の上記有効成分以外の1種以上の有効成分を更に配合してもよい。 The pharmaceutical of the present invention can be administered orally or parenterally, but is preferably administered orally. When the pharmaceutical product of the present invention is orally administered, it is formulated or used in tablets (including dragees), capsules, granules, powders, pills, liquids for internal use, suspensions, emulsions, syrups, etc. You may make it the dried product re-dissolved in the case. In addition, when the pharmaceutical product of the present invention is administered parenterally, it is formulated into an injection (eg, subcutaneous injection, intravenous injection, intramuscular injection, intraperitoneal injection), infusion, suppository, etc. In the case of pharmaceutical preparations, they are provided in the form of unit dosage ampoules or multi-dose containers. In formulating, one or more active ingredients other than the active ingredients of the present invention may be further blended.
本発明の医薬品は、有効成分である液化麹が天然物由来であるため、安全性が高く副作用がないため、前述の活性酸素に起因して生じる各種疾患や障害の予防及び/又は治療用医薬として用いることができる。本発明の医薬品は、前述の各疾患の予防及び/又は治療用医薬として用いる場合、ヒト、マウス、ラット、ウサギ、イヌ、ネコ等の哺乳動物に対して経口または非経口的に安全に投与することができる。 The pharmaceutical product of the present invention is a pharmaceutical product for preventing and / or treating various diseases and disorders caused by the above-mentioned active oxygen because the liquefied koji, which is an active ingredient, is derived from a natural product and thus has high safety and no side effects. Can be used as The pharmaceutical of the present invention is safely administered orally or parenterally to mammals such as humans, mice, rats, rabbits, dogs, cats and the like when used as preventive and / or therapeutic drugs for the above-mentioned respective diseases. be able to.
本発明の医薬品の投与量は、疾患の種類、投与対象の年齢、性別、体重、症状の程度、又は投与方法などに応じて適宜決定することができる。通常、成人1日当たり、液化麹として、10mg〜10g、好ましくは50mg〜5g、より好ましくは100mg〜1gとなる量を、一日一回ないし数回投与する。 The dosage of the pharmaceutical agent of the present invention can be appropriately determined according to the type of disease, the age, sex, body weight, symptom level, or administration method of the subject. In general, an amount of 10 mg to 10 g, preferably 50 mg to 5 g, more preferably 100 mg to 1 g as liquefied sputum is administered once a day or several times per day for an adult.
また、本発明の抗酸化剤は、医薬品のみならず、医薬部外品や化粧品にも配合できる。医薬部外品や化粧品としては、皮膚外用組成物が好ましく、水溶液系、可溶化系、乳化系、粉末系、油液系、ゲル系、軟膏系、エアゾール系等のいずれの剤型でもよい。皮膚外用組成物の種類としては、例えば、化粧水、乳液、ジェル、美容液、一般クリーム、日焼け止めクリーム、パック、マスク、洗顔料、化粧石鹸、ファンデーション、おしろい、浴用剤、ボディローション、ボディシャンプー等が挙げられるが、これらに限定はされない。 Moreover, the antioxidant of this invention can be mix | blended with not only a pharmaceutical but a quasi-drug and cosmetics. As a quasi-drug or cosmetic, a composition for external application to the skin is preferable, and any dosage form such as an aqueous solution system, a solubilization system, an emulsification system, a powder system, an oil-liquid system, a gel system, an ointment system, and an aerosol system may be used. Examples of the composition for external use of skin include, for example, lotion, milky lotion, gel, cosmetic liquid, general cream, sunscreen cream, pack, mask, facial cleanser, cosmetic soap, foundation, funny, bath preparation, body lotion, body shampoo. However, it is not limited to these.
上記医薬部外品や化粧品は、皮膚外用組成物において通常使用されている各種成分、添加剤、基剤等をその種類に応じて選択し、適宜配合し、当分野で公知の手法に従って製造することができる。その形態は、液状、乳液状、クリーム状、ゲル状、ペースト状、スプレー状等のいずれであってもよい。 The above quasi-drugs and cosmetics are produced by selecting various ingredients, additives, bases and the like that are usually used in the composition for external use according to the type, blending them appropriately, and producing them according to methods known in the art. be able to. The form may be liquid, emulsion, cream, gel, paste, spray or the like.
本発明の医薬部外品または化粧品における液化麹の配合量は、所望の抗酸化作用が発揮できる量であればよいが、対象製品の一般的な使用量、製品の形態、効能・効果、及びコストなどを考慮して適宜設定すればよい。 The amount of liquefied koji in the quasi-drug or cosmetic of the present invention may be any amount that can exert the desired antioxidant effect, but the general usage amount of the target product, the form of the product, the efficacy and effects, and What is necessary is just to set suitably in consideration of cost.
さらに、麹または上記の加熱処理により得られる液化麹は、学習・記憶能向上作用、脳のアミロイドβ蓄積量の減少作用をも有する。したがって、麹または液化麹は、認知症の治療及び/又は予防剤として使用できる。 Furthermore, sputum or liquefied sputum obtained by the above heat treatment also has an effect of improving learning / memory ability and a function of reducing the amount of amyloid β accumulated in the brain. Therefore, sputum or liquefied sputum can be used as an agent for treating and / or preventing dementia.
本明細書において「認知症」は、脳の細胞が損傷されることにより現れる記憶障害、見当識障害、行動障害、理解・判断力障害、実行機能障害などの中核症状を伴う疾患をいう。認知症は、大きく「アルツハイマー型認知症」と「脳血管性認知症」に分けられるが、本発明における「認知症」は、これらのいずれをも含み、その原因となる疾患や障害は問わない。例えば、アルツハイマー病、(びまん性)レビー小体病、パーキンソン病、前頭側頭型認知症(意味性認知症、進行性非流暢性失語、進行性核上性麻痺など)、ハンチントン病等の変性性認知症;多発梗塞性認知症広範虚血型、多発脳梗塞型、限局性脳梗塞型、遺伝性血管性認知症(CADASILなど)等の血管性認知症;クロイツフェルト・ヤコブ病、HIV関連認知症;慢性硬膜下血腫、正常圧水頭症、甲状腺機能低下症に起因する認知症などが挙げられるが、これらに限定されない。本発明の認知症を治療及び/又は予防剤は、特に加齢により増加する酸化ストレスに起因するアルツハイマー病や老化に伴う認知症の改善に有効である。 As used herein, “dementia” refers to a disease accompanied by core symptoms such as memory impairment, disorientation, behavioral disorder, understanding / judgment disorder, and executive dysfunction that appear when brain cells are damaged. Dementia is broadly divided into “Alzheimer's dementia” and “cerebral vascular dementia”, and “dementia” in the present invention includes any of these, and any disease or disorder that causes it is not considered. . For example, Alzheimer's disease, (diffuse) Lewy body disease, Parkinson's disease, frontotemporal dementia (semantic dementia, progressive non-fluent aphasia, progressive supranuclear paralysis, etc.), degeneration such as Huntington's disease Vascular dementia: multiple ischemic dementia, extensive ischemia, multiple cerebral infarction, localized cerebral infarction, hereditary vascular dementia (CADASIL, etc.); Creutzfeldt-Jakob disease, HIV-related cognition Diseases include, but are not limited to, chronic subdural hematoma, normal pressure hydrocephalus, dementia due to hypothyroidism, and the like. The agent for treating and / or preventing dementia of the present invention is particularly effective in improving Alzheimer's disease caused by oxidative stress that increases with aging and dementia associated with aging.
本明細書において、認知症の治療とは、認知症を治癒すること、認知症に伴う各症状の緩和および進行を抑制することをいい、認知症の予防とは、認知症の発症を抑制することおよび発症を遅延させるをいう。 In this specification, the treatment of dementia refers to curing dementia, suppressing the alleviation and progression of each symptom associated with dementia, and prevention of dementia refers to suppressing the onset of dementia. And delaying onset.
本発明の認知症の治療及び/又は予防剤は、前記抗酸化剤と同様に、そのまま用いることも可能であるが、本発明の効果を損なわない範囲で適宜他の成分を配合し、医薬品や飲食品の形態で使用することができる。 The therapeutic and / or prophylactic agent for dementia of the present invention can be used as it is, as in the case of the antioxidant. However, other ingredients are appropriately blended within a range not impairing the effects of the present invention, It can be used in the form of food and drink.
本発明の認知症の治療及び/又は予防剤を医薬品とする場合は、医薬上許容される担体等の添加剤を必要に応じて組み合わせて製剤化する。医薬品の投与経路、剤型、添加剤の種類、投与量は、前記の抗酸化剤の場合と同様である。 When the therapeutic and / or prophylactic agent for dementia of the present invention is used as a pharmaceutical, it is formulated by combining additives such as a pharmaceutically acceptable carrier as necessary. The administration route, dosage form, type of additive, and dosage of the pharmaceutical agent are the same as in the case of the antioxidant.
また、飲食品とする場合も、食品として許容される他の材料と必要に応じて組み合わせて調製する。飲食品の形態や種類、添加物の種類、飲食品への配合量は、前記の抗酸化剤の場合と同様である。本発明の飲食品には、その本体、包装、説明書、宣伝物に効能の表示、例えば、抗酸化作用を有する旨の表示、認知症の改善および予防の表示などが付されたものであってもよい。 Moreover, also when setting it as food-drinks, it combines and prepares with the other material accept | permitted as food as needed. The form and kind of food / beverage products, the kind of additive, and the compounding quantity to food / beverage products are the same as that of the said antioxidant. The food / beverage product of the present invention has its main body, packaging, instructions, promotional material with indications of efficacy, for example, an indication that it has an antioxidant effect, an indication of improvement and prevention of dementia, etc. May be.
以下、実施例によって本発明を更に具体的に説明するが、これらの実施例は本発明を限定するものでない。
(実施例1)液化麹の調製(一段階法)
(1) 製麹
精白米1.5kgを洗米後、水に1時間浸漬し、水切り後50分蒸煮した。その後40℃まで冷却して、1.5gの白麹種麹を蒸米に撒き、よく混合し、湿度90%、35℃の恒温器内で40時間かけて製麹した。この間、麹の温度が40℃を超えないように攪拌や冷却を繰り返して管理した。できあがった麹は凍結乾燥後、粉砕し保存した。
EXAMPLES Hereinafter, the present invention will be described more specifically with reference to examples, but these examples do not limit the present invention.
(Example 1) Preparation of liquefied soot (one-step method)
(1) Koji making After washing 1.5 kg of polished rice, it was immersed in water for 1 hour, and drained and steamed for 50 minutes. Thereafter, the mixture was cooled to 40 ° C., 1.5 g of white birch seed koji was sown in steamed rice, mixed well, and kneaded in a thermostat at 90% humidity and 35 ° C. for 40 hours. During this period, stirring and cooling were repeated and controlled so that the temperature of the soot did not exceed 40 ° C. The finished koji was lyophilized, crushed and stored.
(2) 加熱処理
この乾燥粉末麹2gに水を2ml加え、スクリューキャップ付きのチューブに入れ、水分が蒸発しないよう密栓し、45℃、55℃、65℃、または75℃の恒温水槽で7日間の加熱処理を行い、液化麹(一段階法)を得た。
加熱処理して得られた上記液化麹4g、または加熱処理をしていない乾燥粉末麹(未加熱麹)2gを脱イオン水3mlに加え、ホモジナイズした後、ホモジネートを遠心分離(4800×g、5分)し、得られた沈殿物に脱イオン水5mlを加え、再懸濁したものをさらに遠心分離(4800×g、5分)し、上清を合わせて分析用の試料とした。
(2)
4 g of the above-mentioned liquefied soot obtained by heat treatment or 2 g of dry powder soot not heated (unheated soot) is added to 3 ml of deionized water and homogenized, and then the homogenate is centrifuged (4800 × g, 5 5 ml of deionized water was added to the resulting precipitate, and the resuspended product was further centrifuged (4800 × g, 5 minutes), and the supernatant was combined into a sample for analysis.
(3) 褐変と液化の程度の確認
55℃加熱処理麹について、加熱処理開始から1,3,5,7日目に褐変と液化の程度を調べた。褐変の程度は400nmおける吸光度で評価し、吸光度は試料を適宜脱イオン水で希釈し、UV/VIS Spectrophotometer (Shimadzu UV-1700, Kyoto, Japan)を用いて測定した。また、液化の程度は粘度で評価した。粘度は、Rapid Visco Analyser(Newpoint Scientific Pty., Ltd., NSW, Australia)で測定し、Rapid Visco Unit(RVU)で示した。結果を図1に示す。400nmにおける吸光度の増加は3日目から始まり、5日目、7日目で急激に上昇した。この結果は、メイラード反応が加熱中に進行したことを示唆した。また、最初固体状態であった麹の粘度は3日目以降急激に減少した。この結果は、加熱により酵素反応による液化が進行したことを示唆した。
(3) Confirmation of degree of browning and liquefaction The degree of browning and liquefaction was examined on the 1st, 3rd, 5th and 7th days from the start of the heat treatment for the 55 ° C heat-treated rice cake. The degree of browning was evaluated by absorbance at 400 nm, and the absorbance was measured using a UV / VIS Spectrophotometer (Shimadzu UV-1700, Kyoto, Japan) after appropriately diluting the sample with deionized water. The degree of liquefaction was evaluated by viscosity. Viscosity was measured with Rapid Visco Analyzer (Newpoint Scientific Pty., Ltd., NSW, Australia) and indicated in Rapid Visco Unit (RVU). The results are shown in FIG. The increase in absorbance at 400 nm began on the third day and increased rapidly on the fifth and seventh days. This result suggested that the Maillard reaction proceeded during heating. In addition, the viscosity of the soot that was initially in a solid state rapidly decreased after the third day. This result suggested that the liquefaction by the enzyme reaction proceeded by heating.
(4) 抗酸化活性の確認
加熱処理開始から7日後の55℃液化麹、75℃液化麹のそれぞれについて、スーパーオキシドラジカル消去能(SOSA)及び酸素ラジカル吸収能(ORAC)を測定した。SOSAはCLETA-S antioxidant activity assay kit (ATTO CO, Tokyo Japan)、ORACは、OxiSelectTM ORAC activity assay kit (Cell Biolabs, Inc., San diego, CA, USA)をそれぞれ用い、キットの指示書に従って測定した。
(4) Confirmation of antioxidant activity Superoxide radical scavenging ability (SOSA) and oxygen radical absorption ability (ORAC) were measured for 55 ° C. liquefied soot and 75 ° C. liquefied
その結果、加熱前の麹に比べて、55℃加熱ではSOSAが4.9倍、ORACが4.3倍、75℃加熱ではSOSAが20倍、ORACが3.5倍に上昇し、加熱が麹の抗酸化活性に有効であることが示された(後記表1)。また、加熱温度(45〜75℃)とSOSAまたはORACとの関係を調べたところ、SOSAは温度依存的に高い値を示したが、ORACは加熱温度に依らずほぼ同じ値を示すことも確認した。 As a result, SOSA increased by 4.9 times, ORAC increased by 4.3 times, heated by 75 ° C, SOSA increased by 20 times, and ORAC increased by 3.5 times when heated at 55 ° C, compared to the unheated soot, and heating increased the antioxidant activity of the soot. It was shown to be effective (Table 1 below). In addition, when the relationship between the heating temperature (45 to 75 ° C.) and SOSA or ORAC was examined, SOSA showed a high value depending on the temperature, but it was also confirmed that ORAC showed almost the same value regardless of the heating temperature. did.
(5) グルコース含量とアミノ窒素
55℃加熱処理麹、75℃加熱処理麹のそれぞれについて、グルコース含量とアミノ窒素含量の経時的変化を比較した。結果を図2に示す。55℃加熱処理麹では、グルコース含量とアミノ窒素含量ともに1〜2日目で急激に上昇しプラトーに達した。これに対し、75℃加熱処理麹では、55℃加熱処理麹に比べグルコース含量の増加に効果的ではなく、1〜2日を経過後もグルコース含量とアミノ窒素含量ともに変化はなかった。これらの結果から、酵素反応は55℃2日目で終了すること、75℃加熱では、酵素反応で生じたグルコースやアミノ酸を基質としてメイラード反応が進行すると考えられた。
(5) Glucose content and amino nitrogen The changes over time of the glucose content and amino nitrogen content were compared for each of the 55 ° C. heat-treated rice cake and the 75 ° C. heat-treated rice cake. The results are shown in FIG. In the 55 ° C. heat-treated rice cake, both the glucose content and the amino nitrogen content rapidly increased on the first or second day and reached a plateau. In contrast, the 75 ° C. heat-treated rice bran was not effective in increasing the glucose content as compared with the 55 ° C. heat-treated rice cake, and neither the glucose content nor the amino nitrogen content changed after 1 to 2 days. From these results, it was considered that the enzymatic reaction is completed at 55 ° C. on the second day, and that heating at 75 ° C. causes the Maillard reaction to proceed using glucose and amino acids generated by the enzymatic reaction as substrates.
(実施例2)液化麹の調製(二段階法)
実施例1の結果から、55℃で酵素反応を進行させた後に、より高温で加熱することによりメイラード反応をさらに促進できると考えた。そこで、55℃で2日間加熱処理した後、続いて75℃で5日間加熱処理する二段階法を試みた。
(Example 2) Preparation of liquefied soot (two-stage method)
From the results of Example 1, it was considered that the Maillard reaction could be further promoted by heating at a higher temperature after the enzyme reaction was allowed to proceed at 55 ° C. Therefore, a two-stage method was attempted in which heat treatment was performed at 55 ° C. for 2 days, and then heat treatment was performed at 75 ° C. for 5 days.
(1) 加熱処理
実施例1と同様にして製麹し、調製した乾燥粉末麹2gに水を2ml加え、スクリューキャップ付きのチューブに入れ、水分が蒸発しないよう密栓し、55℃の恒温水槽で2日間加熱処理し、その後、75℃の恒温水槽で5日間の加熱処理を行い、液化麹(二段階法)を得た。
(1) Heat treatment As in Example 1, 2 ml of water is added to 2 g of the dried powder koji prepared, put into a tube with a screw cap, tightly sealed so that moisture does not evaporate, and kept in a constant temperature water bath at 55 ° C. Heat treatment was performed for 2 days, and then heat treatment was performed for 5 days in a 75 ° C. constant temperature water bath to obtain a liquefied soot (two-stage method).
(2) 抗酸化活性の確認
得られた液化麹(二段階法)について、実施例1と同様にしてSOSAおよびORACを測定した。結果を、実施例1の液化麹(一段階法)と合わせて下記表1に示す。
(2) Confirmation of antioxidant activity The obtained liquefied soot (two-stage method) was measured for SOSA and ORAC in the same manner as in Example 1. The results are shown in Table 1 below together with the liquefied soot of Example 1 (one-step method).
+: Student's t-testにより55℃加熱麹と有意差あり(p<0.05)
++:Student’s t-testにより75℃加熱麹と有意差あり(p<0.05)
+: According to Student's t-test, there is a significant difference from 55 ° C heating pad (p <0.05)
++: Significantly different from Student's t-test with 75 ° C heating (p <0.05)
その結果、SOSAが未加熱麹の93.8倍、ORACが5.9倍に増強された。この値は55℃一定、または75℃一定温度で7日間加熱処理した麹よりも高く、麹の抗酸化活性増強には高温でのメイラード反応に先立ち、酵素反応の促進を図ることが効果的であることがわかった。 As a result, SOSA was increased 93.8 times and ORAC 5.9 times that of unheated soot. This value is higher than that of cocoon heat-treated for 7 days at a constant temperature of 55 ° C or 75 ° C. To enhance the antioxidant activity of cocoon, it is effective to promote the enzyme reaction prior to the Maillard reaction at high temperature. I found out.
さらに、抗酸化活性を有する化合物として米中の総フェノール含量(TPC)及びメイラード反応生成物である5−ヒドロキシメチルフルフラール(HMF)に着目し、未加熱麹と上記の二段階法による加熱処理後の麹について、総フェノール含量(TPC)をFolin Ciocalteu法により測定し、5−ヒドロキシメチルフルフラール(HMF)含量をHPLCにて分析した。図3Aに、TPCおよびHMFの経時変化、また、図3Bに、SOSAおよびORACの経時変化を示す。加熱処理麹(二段階法)では、TPCが1.28から8.46(mgGAE/g)に、HMFは非検出から28.6mg/gに増加した(図3A)。よって上記条件で加熱処理した麹の抗酸化活性は、フェノール類やHMFを始めとしたメイラード反応物に由来していることが示唆された。 In addition, paying attention to the total phenol content (TPC) in rice and Maillard reaction product 5-hydroxymethylfurfural (HMF) as a compound having antioxidant activity, after heat treatment by unheated rice bran and the above two-step method The total phenol content (TPC) was measured by Folin Ciocalteu method and the 5-hydroxymethylfurfural (HMF) content was analyzed by HPLC. FIG. 3A shows changes over time in TPC and HMF, and FIG. 3B shows changes over time in SOSA and ORAC. In the heat-treated bowl (two-stage method), TPC increased from 1.28 to 8.46 (mgGAE / g), and HMF increased from non-detected to 28.6 mg / g (FIG. 3A). Therefore, it was suggested that the antioxidant activity of the soot heat-processed on the said conditions originates in the Maillard reaction material including phenols and HMF.
また、麹の種類による抗酸化活性の違いについて白麹と黄麹で比較した。白麹種麹または黄麹種麹を用いて製麹した麹を上記の二段階法による加熱処理を行った。得られた液化麹について、SOSAおよびORACを測定し、また、TPCおよびHMFについても調べた。結果を下記表2に示す。 In addition, the difference in antioxidant activity depending on the type of cocoon was compared between white rabbit and jaundice. The koji made from white koji seed koji or yellow koji seed koji was heat-treated by the above two-stage method. The obtained liquefied soot was measured for SOSA and ORAC, and also for TPC and HMF. The results are shown in Table 2 below.
**:Student's t-testにより液化麹(白)と有意差あり(p<0.01)
**: Significantly different from liquefied lees (white) by Student's t-test (p <0.01)
いずれも優れた抗酸化活性が得られたが、HMFの生産性の点では白麹のほうが優れていた。 In both cases, excellent antioxidant activity was obtained, but white cocoon was superior in terms of HMF productivity.
(実施例3)
本実施例において、統計解析にはIBM SPSS Statistics version 19 を用い、2群間の検定は Student's t-test、多重検定は One-way ANOVA Tukey法で行った。また、本実験は鹿児島大学動物実験委員会で承認された後 (第A10053号)、動物の愛護及び管理に関する法律 (動物愛護法,平成17年6月22日公布,平成18年6月1日施行,環境省) を遵守して行った。
(Example 3)
In this example, IBM SPSS Statistics version 19 was used for statistical analysis, and the test between two groups was performed by Student's t-test, and the multiple test was performed by the One-way ANOVA Tukey method. In addition, after this experiment was approved by the Kagoshima University Animal Experiment Committee (No. A10053), the Act on the Protection and Management of Animals (Animal Protection Act, promulgated on June 22, 2005, June 1, 2006) (Enforcement, Ministry of the Environment)
(1) 実験飼料の調製
実験飼料の基準食にはCE-2(九動株式会社より購入)を用いた。実験食には、麹または実施例2で調製した液化麹を凍結乾燥し、粉末化した麹粉末または液化麹粉末をCE-2に対して0.5%(w/w) にて混合したものを用いた。麹は白麹(Aspergillus kawachii) を用いた。CE-2、麹、および液化麹の栄養組成を表3に示す。
(1) Preparation of experimental feed CE-2 (purchased from Kudo Co., Ltd.) was used as the standard diet for the experimental feed. For the experimental meal, use the koji or liquefied koji prepared in Example 2 freeze-dried, and mix the powdered koji powder or liquefied koji powder at 0.5% (w / w) with respect to CE-2. It was. As the cocoon, Aspergillus kawachii was used. Table 3 shows the nutritional composition of CE-2, koji, and liquefied koji.
(2) 実験動物
通常老化作用を示す老化抵抗モデルマウスSAMR1(Senescence Accelarated-Resistant Mice)と、促進老化および短寿命を示し、老化とともに学習・記憶障害を特徴的に発症する老化促進モデルマウスSAMP8(Senescence Accelerated-Prone Mice)の2種類を用いた。
8週齢雄の老化促進モデルマウスSAMP8 (27匹) と、同系統の老化抵抗マウスSAMR1 (12匹) は、日本SLC (株) から購入した。
(2) Experimental animals Aging resistance model mouse SAMR1 (Senescence Accelarated-Resistant Mice), which shows normal aging action, and accelerated aging and short life, SAMP8 ( Senescence Accelerated-Prone Mice) was used.
8-week-old male aging-promoting model mice SAMP8 (27 mice) and aging-resistant mice SAMR1 (12 mice) of the same strain were purchased from Japan SLC.
(3) 給餌およびモリス水迷路実験のスケジュール
マウスの給餌およびモリス水迷路実験のスケジュールを図4に示す。1週間の予備飼育後、SAMP8マウスの群分けおよび泳ぎに異常のあるマウスを確認するための水迷路予備実験を5日間行い、SAMP8マウスを遊泳速度が均等になるよう9匹ずつ3群に分けた。飼育開始から実験食の給餌を開始するまでの2週間を予備飼育期間とした。
(3) Schedule of feeding and Morris water maze experiment The schedule of mouse feeding and Morris water maze experiment is shown in FIG. After 1 week of pre-breeding, SAMP8 mice were grouped and a water maze preliminary experiment to confirm mice with abnormal swimming was conducted for 5 days, and SAMP8 mice were divided into 3 groups of 9 mice with equal swimming speed. It was. Two weeks from the start of the breeding to the start of feeding the experimental food was designated as a preliminary breeding period.
コントロールであるSAMR1マウスには飼育開始から屠殺時まで終始通常(基準)食であるCE-2を与えた (SAMR1/CE-2群)。実験食群であるSAMP8マウスには、予備飼育期間中CE-2を与えた後、Sharm controlとして引き続きCE-2を与える群 (SAMP8/CE-2群)、CE-2と麹粉末を与える群(SAMP8/CE-2+麹群)、CE-2と液化麹粉末を与える群 (SAMP8/CE-2+液化麹群)の3群(9 匹/群)に分け、予備飼育期間後それぞれの実験食で7週間の本飼育を行った。 The control SAMR1 mice were given CE-2, the normal (reference) diet, from the start of breeding to the time of sacrifice (SAMR1 / CE-2 group). SAMP8 mice in the experimental diet group were given CE-2 during the pre-breeding period, then continued to receive CE-2 as a Sharm control (SAMP8 / CE-2 group), a group given CE-2 and sputum powder (SAMP8 / CE-2 + 麹 group), CE-2 and liquefied cocoon powder group (SAMP8 / CE-2 + liquefied cocoon group) divided into 3 groups (9 animals / group). The food was raised for 7 weeks.
マウスはゲージにて1匹ずつ飼育し、気温20℃、12時間/12時間 (8:00〜20:00) の明暗周期とした。自由給餌及び自由飲水で飼育した。実験食給餌開始から3週間後 (13週齢)にモリス水迷路本実験を行った。17週齢時に飼育を終了し、エーテル麻酔下で心採血により全血を採取し安楽死させた。 Mice were housed one by one with a gauge, and the temperature was 20 ° C, and the light-dark cycle was 12 hours / 12 hours (8: 00-20: 00). Breeding with free feeding and free drinking. A Morris water maze experiment was conducted 3 weeks after the start of feeding the experimental diet (13 weeks of age). Breeding was terminated at the age of 17 weeks, and whole blood was collected by cardiac blood sampling under ether anesthesia and euthanized.
(4) モリス水迷路実験
実験食給餌開始3週間後にモリス水迷路本実験を行い、行動学的にマウスの学習・記憶能力を判定した。
(4) Morris water maze experiment The Morris water maze experiment was conducted 3 weeks after the start of experimental diet feeding, and the learning and memory ability of the mice was determined behaviorally.
直径120cm、高さ20cmの円形プール(図5A)を部屋の隅に設置し、外環境からの位置情報をなるべく低減させた。ゴールとなるプラットホーム (PF)には透明のアクリル樹脂を用い、4つに等分割したプールの右上領域(図5B)に設置し、水面下に投入すると水面から察知できないように水中に沈めた(図5A)。プールの水温は25℃にした。実験はトレーニング期間とプローブテストの2種類の実験段階に分けて行った。トレーニング期間では、PFを沈めていない3つの領域より壁面に頭を向けた状態でマウスを投入し、右上領域に設置したPFに辿り着くまでに経過した時間をそれぞれ測定した。PFに到達したマウスはそのままPF上に15秒間滞在させ位置を学習させた後にケージに戻した。60秒を超えてもPFに辿り着けなかった場合その時点で遊泳を終了し、到達時間を60秒とした上で強制的にPF上へ移動させ、15秒間滞在させた。この操作を1日3回、14日間毎日行い、その翌日(15日目)にPFを撤去したプローブテストを行った。プローブテストではPFを撤去した条件でマウスを60秒間自由遊泳させ、PF周辺領域の滞在時間を測定した。 A circular pool (FIG. 5A) with a diameter of 120 cm and a height of 20 cm was installed at the corner of the room to reduce position information from the outside environment as much as possible. The goal platform (PF) is made of transparent acrylic resin, installed in the upper right area of the pool (Fig. 5B) divided into four equal parts and submerged in water so that it cannot be detected from the water surface when thrown under the surface of the water ( FIG. 5A). Pool water temperature was 25 ° C. The experiment was divided into two types of experimental stages: training period and probe test. During the training period, mice were placed with their heads facing the walls from the three areas where the PF was not submerged, and the time elapsed until reaching the PF placed in the upper right area was measured. Mice that reached the PF stayed on the PF for 15 seconds to learn the position and then returned to the cage. If the PF could not be reached even after 60 seconds, the swimming was terminated at that point, and after reaching the arrival time of 60 seconds, it was forcibly moved to the PF and stayed for 15 seconds. This operation was performed 3 times a day for 14 days every day, and the probe test was performed on the next day (15th day) with PF removed. In the probe test, the mice were allowed to swim freely for 60 seconds under the condition where the PF was removed, and the residence time in the area around the PF was measured.
トレーニング期間の結果では、SAMP8/CE-2+麹群およびSAMP8/CE-2+液化麹群において、12日目以降では有意差はなかったものの、SAMP8/CE-2群(Sharm control)よりもPF到達時間が短くなり、学習・記憶能力低下が抑制されている傾向が見られ、最終日の14日では有意に早く到達した(図6)。 As a result of the training period, the SAMP8 / CE-2 + group and the SAMP8 / CE-2 + liquefied group had no significant difference after the 12th day, but PF than the SAMP8 / CE-2 group (Sharm control). The arrival time was shortened and the decline in learning / memory ability was suppressed, reaching significantly earlier on the 14th day (FIG. 6).
プローブテストの結果では、有意差はなかったものの、SAMP8/CE-2群(Sharm control)に比べSAMP8/CE-2+麹群およびSAMP8/CE-2+液化麹群においてPF周辺領域滞在時間が増加する傾向にあった(図7)。 Although there was no significant difference in the results of the probe test, the residence time around the PF area was increased in the SAMP8 / CE-2 + group and SAMP8 / CE-2 + liquefied group compared to the SAMP8 / CE-2 group (Sharm control) (Fig. 7).
(5) アミロイド線維蓄積の検出
マウスの脳組織中に蓄積されたアルツハイマーの原因とされるアミロイドβ量を免疫蛍光染色により測定した。実験には海馬の組織切片を用い、測定部位は記憶形成において重要な海馬DG、CA1およびCA3領域の三つとした。
(5) Detection of amyloid fibril accumulation The amount of amyloid β caused by Alzheimer accumulated in brain tissue of mice was measured by immunofluorescence staining. Hippocampal tissue sections were used in the experiment, and the measurement sites were the hippocampal DG, CA1, and CA3 regions important for memory formation.
ホルマリン固定組織をパラフィン包埋し、6 μmの薄片を作成した。脱パラフィン後、Proteostat Amyloid Plaque Detection Kit (Enzo, Plymouth Meeting, PA) を用い、添付の説明書に準じてアミロイドオリゴマーを染色した。細胞核の像を得るためにDNAをDAPI染色した。レーザー共焦点顕微鏡で撮影後、海馬CA1,CA3,DG領域の蛍光強度をImage J Software (NIH,Bethesda,MD) を用いて数値化した。 Formalin-fixed tissue was embedded in paraffin to produce 6 μm slices. After deparaffinization, amyloid oligomers were stained using Proteostat Amyloid Plaque Detection Kit (Enzo, Plymouth Meeting, PA) according to the attached instructions. DNA was stained with DAPI to obtain an image of the cell nucleus. After photographing with a laser confocal microscope, the fluorescence intensity in the hippocampal CA1, CA3, and DG regions was quantified using Image J Software (NIH, Bethesda, MD).
その結果、CA1領域においては各群間に有意な差は認められなかったが、DGおよびCA3領域においてSAMP8/CE-2群(Sharm control)に比べSAMP8/CE-2+麹群およびSAMP8/CE-2+液化麹群には有意にアミロイドβ蓄積の減少が認められた(図8)。 As a result, there was no significant difference between the groups in the CA1 region, but in the DG and CA3 regions, compared to the SAMP8 / CE-2 group (Sharm control), the SAMP8 / CE-2 + 麹 group and the SAMP8 / CE group. A significant decrease in amyloid β accumulation was observed in the −2+ liquefied cocoon group (FIG. 8).
以上の結果より、麹および液化麹には認知症の改善および発症予防効果があることが明らかになった。 From the above results, it was revealed that wrinkles and liquefied wrinkles have an effect of improving dementia and preventing onset.
本発明は食品(特にサプリメントなどの機能性飲食品)、医薬品、医薬部外品、化粧品の製造分野において利用できる。 The present invention can be used in the field of manufacturing foods (particularly functional foods and beverages such as supplements), pharmaceuticals, quasi drugs, and cosmetics.
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JP2014240361A (en) * | 2013-06-11 | 2014-12-25 | 丸善製薬株式会社 | Anti aging agent, whitening agent, skin cosmetic, and food and drink |
WO2015102095A1 (en) * | 2014-01-04 | 2015-07-09 | 靖幸 山田 | Fusion of japanese and european traditional food science |
JP2015140325A (en) * | 2014-01-30 | 2015-08-03 | 福岡県 | Collagenase inhibitor, and cosmetic composition and functional food which contain collagenase inhibitor |
JP2018068293A (en) * | 2016-10-27 | 2018-05-10 | 宝酒造株式会社 | Mirin, method for producing the same, and method for producing processed food |
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JP2002330723A (en) * | 2001-02-28 | 2002-11-19 | Samjo Celltech Ltd | Noodle using soybean embryo and method for producing the same and hishios produced by using the same |
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Cited By (6)
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JP2014240361A (en) * | 2013-06-11 | 2014-12-25 | 丸善製薬株式会社 | Anti aging agent, whitening agent, skin cosmetic, and food and drink |
WO2015102095A1 (en) * | 2014-01-04 | 2015-07-09 | 靖幸 山田 | Fusion of japanese and european traditional food science |
JP2015140325A (en) * | 2014-01-30 | 2015-08-03 | 福岡県 | Collagenase inhibitor, and cosmetic composition and functional food which contain collagenase inhibitor |
JP2018068293A (en) * | 2016-10-27 | 2018-05-10 | 宝酒造株式会社 | Mirin, method for producing the same, and method for producing processed food |
JP2022010085A (en) * | 2016-10-27 | 2022-01-14 | 宝酒造株式会社 | Mirin, production metho of mirin, and production method of processed food |
JP7213327B2 (en) | 2016-10-27 | 2023-01-26 | 宝酒造株式会社 | Mirin, method for producing mirin, and method for producing processed food |
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