JP4762987B2 - Antihypertensive agent obtained by lactic acid bacteria culture - Google Patents
Antihypertensive agent obtained by lactic acid bacteria culture Download PDFInfo
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Description
本発明は、特定の培養液中で乳酸菌を培養した培養物を有効成分とする血圧低下剤に関する。 The present invention relates to an antihypertensive agent comprising a culture obtained by culturing lactic acid bacteria in a specific culture solution as an active ingredient.
スピルリナ又はクロレラを含む食品は、従来、緑黄色野菜に特有の栄養成分や、固有の栄養成分を豊富に含むことから、通常の食生活において不足しがちな栄養成分を手軽に摂取できる食品として利用されている。最近、スピルリナ又はクロレラ中で乳酸菌を培養してスピルリナやクロレラの特有の匂いや風味を改善した食品が提案されている（特許文献１及び２参照）。
ところで、特定の健康食品が高血圧症に有効であることがいわれており、乳類に乳酸菌を接種する方法（特許文献３）、乳製品に酵素、次いで乳酸菌を接種する方法（特許文献４）、或いは、糠、胚芽を含む米糠等を一定条件下で水に浸漬する方法（特許文献５）によるものが知られている。
しかし、スピルリナ又はクロレラと乳酸菌とからなる混合体を、水の存在下に維持し、乳酸菌培養を行うことにより得られる培養物が健康食品として用いられるものの、高血圧症に有効であることについてはまったく知られていない。
By the way, it is said that specific health foods are effective for hypertension, a method of inoculating lactic acid bacteria in milk (Patent Document 3), a method of inoculating enzymes in milk products and then lactic acid bacteria (Patent Document 4), Or the method by the method (patent document 5) of immersing the rice bran etc. containing a rice bran and a germ in water under a fixed condition is known.
However, although a mixture obtained from spirulina or a mixture of chlorella and lactic acid bacteria in the presence of water and cultivating lactic acid bacteria is used as a health food, it is completely effective for hypertension. unknown.
本発明の課題は、微細藻類と乳酸菌とを含有する培養液中で、乳酸菌を培養して得られる乳酸菌培養物を有効成分とする血圧低下剤を提供することにある。 The subject of this invention is providing the blood pressure lowering agent which uses as an active ingredient the lactic acid bacteria culture obtained by culture | cultivating lactic acid bacteria in the culture solution containing a micro algae and lactic acid bacteria.
本発明者らは、上記課題を解決するため、微細藻類と乳酸菌とを、水の存在下に維持し、該混合体中の乳酸菌を種々の条件により培養して得られる乳酸菌培養物に対して、血圧低下作用について試験を行ったところ、該培養物は、非常に優れた作用を有することを見出し、本発明を完成するに至った。 In order to solve the above problems, the present inventors maintain a microalgae and lactic acid bacteria in the presence of water, and a lactic acid bacteria culture obtained by culturing lactic acid bacteria in the mixture under various conditions. As a result of a test on blood pressure lowering action, the culture was found to have a very excellent action, and the present invention was completed.
すなわち、本発明は、スピルリナ、クロレラ、又はスピルリナ及びクロレラの混合物と、乳酸菌とを含有する培養液中で、乳酸菌を培養して得られる乳酸菌培養物を有効成分とする血圧低下剤であって、前記乳酸菌培養物の乾燥重量１ｇあたり、２．１ｍｇ以上のγ−アミノ酪酸を含有する血圧低下剤を提供するものである。 That is, the present invention is Spirulina, Chlorella, or Spirulina and mixtures Chlorella, in a culture medium containing lactic acid bacteria, a blood pressure lowering agent comprising as an active ingredient a lactic acid bacterium culture obtained by culturing lactic acid bacteria, The present invention provides a blood pressure lowering agent containing 2.1 mg or more of γ-aminobutyric acid per 1 g of dry weight of the lactic acid bacteria culture .
本発明によれば、微細藻類と乳酸菌とを含有する培養液中で、乳酸菌を培養して得られる乳酸菌培養物を有効成分とする血圧低下剤を得ることができる。 ADVANTAGE OF THE INVENTION According to this invention, the blood pressure lowering agent which uses as an active ingredient the lactic acid bacteria culture obtained by culture | cultivating lactic acid bacteria in the culture solution containing a micro algae and lactic acid bacteria can be obtained.
以下に、本発明の内容を詳細に説明する。
本願発明で用いる微細藻類としては、スピルリナ、クロレラなどが挙げられる。
本発明で用いられるスピルリナとしては、例えば以下のものが挙げられる。
スピルリナ（Ｓｐｉｒｕｌｉｎａ）とは、藍藻類（Ｃｙａｎｏｂａｃｔｅｒｉａ）に包含され、従来一括してスピルリナ属と呼称されていたアルスロスピラ属（Ａｒｔｈｒｏｓｐｉｒａ）及びスピルリナ属（Ｓｐｉｒｕｌｉｎａ）に属する微細な単細胞微生物であり、例えばアルスロスピラ・プラテンシス（Ａｒｔｈｒｏｓｐｉｒａ ｐｌａｔｅｎｓｉｓ）、アルスロスピラ・マキシマ（Ａｒｔｈｒｏｓｐｉｒａ ｍａｘｉｍａ）、アルスロスピラ・ゲイトレリ（Ａｒｔｈｒｏｓｐｉｒａ ｇｅｉｔｌｅｒｉ）、アルスロスピラ・サイアミーゼ（Ａｒｔｈｒｏｓｐｉｒａ ｓｉａｍｅｓｅ）、スピルリナ・メイヤー（Ｓｐｉｒｕｌｉｎａ ｍａｊｏｒ）、スピルリナ・サブサルサ（Ｓｐｉｒｕｌｉｎａ ｓｕｂｓａｌｓａ）、等が挙げられるが、中でも、人工的に培養でき、入手が容易なことから、アルスロスピラ・プラテンシス、アルスロスピラ・マキシマ、アルスロスピラ・ゲイトレリ、アルスロスピラ・サイアミーゼが好ましい。The contents of the present invention will be described in detail below.
Examples of the microalgae used in the present invention include spirulina and chlorella.
Examples of spirulina used in the present invention include the following.
Spirulina is a fine unicellular microorganism belonging to the genus Arthrospira and Spirulina, which is included in the cyanobacteria and has been collectively referred to as the genus Spirulina. Arthrospira platensis, Arthrospira maxima, Arthrospira geitleri, Arthrospira sir sul, Sul, Sul But in Artificially it can be cultured, because the easy availability, Arthrospira platensis, Arthrospira maxima, Arthrospira-Geitoreri, the Arthrospira-Saiamize preferred.
本発明で用いられるクロレラとしては、例えば以下のものが挙げられる。
クロレラ（Chlorella）とは、緑藻類（Ｃｈｌｏｒｏｐｈｙｃｅａｅ）、クロレラ属（Ｃｈｌｏｒｅｌｌａ）の藻類であり、入手が容易で、安全性に優れている点で、例えば、クロレラ・ブルガリス（C. vulgaris)、クロレラ ・レギュラリス（Ｃｈｌｏｒｅｌｌａ
ｒｅｇｕｌａｒｉｓ）、クロレラ・ピレノイドーサ(C. pyrenoidosa)、クロレラ・エリプソイデア(C. ellipsidea)等が挙げられる。Examples of the chlorella used in the present invention include the following.
Chlorella is an algae of Chlorophyceae and Chlorella, which is easily available and excellent in safety. For example, Chlorella vulgaris, Chlorella Regularis (Chlorella
regularis), C. pyrenoidosa, C. ellipsidea and the like.
これらの微細藻類は、従来食品として広く用いられてきたものであり、安全性について、問題のないことが確認されている。
これらの微細藻類としては、生の藻体、乾燥藻体、及び機械的処理等の方法により処理した藻体処理物等が挙げられる。
生の藻体は、例えば、水中で培養されたスピルリナ、クロレラを遠心分離、濾過等の方法により収穫して得られる。生の藻体は、培養槽から収穫後そのままの状態で使用することもできるが、水もしくは生理食塩水で洗浄するのが好ましい。
乾燥藻体は、例えば、前記方法で得られた生の藻体を凍結乾燥処理やスプレー乾燥処理したもの等が挙げられる。
機械的処理の方法により処理した藻体処理物は、例えば、生の藻体を超音波照射処理や、ホモゲナイズ等の機械処理を行うことにより得られる。藻体の機械的処理物は、その後に乾燥処理を施しても良い。These microalgae have been widely used as foods in the past, and it has been confirmed that there are no problems with regard to safety.
Examples of these microalgae include raw alga bodies, dried alga bodies, and processed alga bodies processed by a method such as mechanical treatment.
The raw algal bodies are obtained, for example, by harvesting spirulina or chlorella cultured in water by a method such as centrifugation or filtration. Although raw algal bodies can be used as they are after harvesting from the culture tank, they are preferably washed with water or physiological saline.
Examples of the dried algal bodies include those obtained by freeze-drying or spray-drying the raw algal bodies obtained by the above method.
The processed alga body treated by the mechanical treatment method can be obtained, for example, by subjecting raw alga bodies to ultrasonic treatment or mechanical treatment such as homogenization. The mechanically processed product of algal bodies may be subjected to a drying process thereafter.
本発明で用いられる藻体としては、生の藻体であることが、微細藻類の有効成分をより保持していることから、また、味、臭いの点からも、好ましい。
生の藻体は、通常、収穫する際の水の除去程度により、水に懸濁している懸濁状のものや、懸濁状のものに比べ水の含有量が少ないペースト状のものや、ペースト状のものに比べ水の含有量が少ないケーキ状の状態のものがあるが、いずれの状態のものでも使用できる。スピルリナ、クロレラは、懸濁状にした藻体（以後、懸濁液という場合がある。）を用いるのが好ましい。また、乾燥藻体や藻体処理物を用いる場合は、乾燥している状態でも良いし、水を加えて、生の藻体のように懸濁状やペースト状やケーキ状にしたものでも良い。The algal bodies used in the present invention are preferably raw algal bodies because they hold more active ingredients of microalgae, and are also preferable in terms of taste and smell.
Raw alga bodies are usually suspended in water depending on the degree of water removal when harvesting, pastes with less water content than suspended ones, There is a cake-like state in which the water content is less than that in the pasty state, but any state can be used. For Spirulina and Chlorella, it is preferable to use suspended algal bodies (hereinafter sometimes referred to as suspension). Moreover, when using a dry alga body or a processed alga body, it may be in a dry state, or it may be in the form of a suspension, paste or cake like raw alga body by adding water. .
微細藻類は、それらの有する有効成分を損なわないためには加熱殺菌しない方が好ましいが、必要に応じて加熱殺菌したものを用いることもできる。
ここで、微細藻類は、スピルリナ、又はクロレラを単独で用いても、スピルリナとクロレラの混合物を用いてもどちらでもよい。The microalgae are preferably not heat-sterilized in order not to impair their active ingredients, but those that have been heat-sterilized can be used if necessary.
Here, as the microalgae, either spirulina or chlorella may be used alone, or a mixture of spirulina and chlorella may be used.
次に、乳酸菌について説明する。
乳酸菌は、古来、食品の保蔵と調味を目的に発酵乳製品、醸造製品、野菜・果実の漬物など多くの食品の加工に利用されている。本発明で用いる乳酸菌としては、食用として利用できる乳酸菌であれば制限無く用いることができる。乳酸菌としては、由来する生育環境により乳系乳酸菌、植物系乳酸菌、腸管系乳酸菌や、藻類の生育する自然湖に由来する乳酸菌等に分類される。また乳酸菌は、その生育至適条件により中温性菌、高温性菌、耐塩性菌等にも分類されるが、いずれの性質を有する菌でも良い。Next, lactic acid bacteria will be described.
Lactic acid bacteria have been used since long ago for the processing of many foods such as fermented milk products, brewed products, and pickles of vegetables and fruits for the purpose of food preservation and seasoning. As the lactic acid bacteria used in the present invention, any lactic acid bacteria that can be used for food can be used without limitation. Lactic acid bacteria are classified into milk-based lactic acid bacteria, plant-based lactic acid bacteria, intestinal lactic acid bacteria, lactic acid bacteria derived from natural lakes where algae grow, and the like depending on the growth environment from which they are derived. Lactic acid bacteria are classified into mesophilic bacteria, thermophilic bacteria, salt-tolerant bacteria, and the like depending on their optimal growth conditions, but bacteria having any property may be used.
本発明で用いる乳酸菌としては、分類学上、ラクトバチルス（Ｌａｃｔｏｂａｃｉｌｌｕｓ）属、ペディオコッカス（Ｐｅｄｉｏｃｏｃｃｕｓ）属、テトラゲノコッカス（Ｔｅｔｒａｇｅｎｏｃｏｃｃｕｓ）属、カルノバクテリウム（Ｃａｒｎｏｂａｃｔｅｒｉｕｍ）属、バゴコッカス（Ｖａｇｏｃｏｃｃｕｓ）属、ロイコノストック（Ｌｅｕｃｏｎｏｓｔｏｃ）属、ワイセラ（Ｗｅｉｓｓｅｌｌａ）属、オエノコッカス（Ｏｅｎｏｃｏｃｃｕｓ）属、アトポビウム（Ａｔｏｐｏｂｉｕｍ）属、ストレプトコッカス（Ｓｔｒｅｐｔｏｃｏｃｃｕｓ）属（正式名はエンテロコッカス属、本明細書においてはエンテロコッカス属に包含する）、エンテロコッカス（Ｅｎｔｅｒｏｃｏｃｃｕｓ）属、ラクトコッカス（Ｌａｃｔｏｃｏｃｃｕｓ）属、アエロコッカス（Ａｅｒｏｃｏｃｃｕｓ）属、アロイオコッカス（Ａｌｌｏｉｏｃｏｃｃｕｓ）属、メリソコッカス（Ｍｅｌｉｓｓｏｃｏｃｃｕｓ）属、ビフィドバクテリウム（Ｂｉｆｉｄｏｂａｃｔｅｒｉｕｍ）属等が挙げられ、更に、例えばラクトバチルス デルブルエキ（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｄｅｌｂｒｕｅｃｋｉｉ）、ラクトバチルス プランタルム（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｐｌａｎｔａｒｕｍ）、ラクトバチルス アシドフィルス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ａｃｉｄｏｐｈｉｌｕｓ）、ラクトバチルス ブレビス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｂｒｅｖｉｓ）、ラクトコッカス ラクティス（Ｌａｃｔｏｃｏｃｃｕｓ ｌａｃｔｉｓ）、ロイコノストック エスピー（Ｌｅｕｃｏｎｏｓｔｏｃ ｓｐ．）、エンテロコッカス カセリフラブス（Ｅｎｔｅｒｏｃｏｃｃｕｓ ｃａｓｓｅｌｉｆｌａｖｕｓ） 等の種が挙げられる。本発明に用いられる乳酸菌としては、ラクトバチルス属、ラクトコッカス属、エンテロコッカス属 に属する乳酸菌が好ましい。 The lactic acid bacteria used in the present invention include taxonomically, Lactobacillus genus, Pediococcus genus, Tetragenococcus genus, Carnobacterium genus, Vagococcus genus, Vagococcus genus, and Vagococcus genus. The genus Leuconostoc, the genus Weissella, the genus Oenococcus, the genus Atopobium, the genus Streptococcus (official name is the genus Enterococcus, the genus Enterococcus (Enterococcus) genus, Lactococcus genus, Examples include the genus Aerococcus, the genus Alloiococcus, the genus Melissococcus, the genus Bifidobacterium, and the like. plantarum, Lactobacillus acidophilus, Lactobacillus brevis, Lactococcus lactis, Leuconostoc sp. Rokokkasu Kaserifurabusu (Enterococcus casseliflavus) species, and the like. The lactic acid bacteria used in the present invention are preferably lactic acid bacteria belonging to the genus Lactobacillus, Lactococcus, and Enterococcus.
乳酸菌は、単独種で使用しても良いし、２種類以上の菌を混合して使用しても良い。また、後述する培養工程において、同じ種類の菌を２段階以上に分けて植菌して培養しても良いし、異なった菌種を２段階以上に分けて植菌し培養しても良い。
乳酸菌は、寒天培地や液体培地で培養後、冷蔵保存、凍結保存、乾燥保存等の保存方法により保存しておいたものを用いても良いが、これらの保存しておいた乳酸菌を液体培地に植菌して培養したもの（以下、種培養液と略記する。）を用いるのが乳酸菌の増殖速度が速く、アセトアルデヒド、ジアセチル等のフレーバー類の産生能、有機酸産生能等の活性が高いことから好ましい。種培養液を培養するのに用いる培地は、用いる乳酸菌が生育可能な培地であれば良く制限はないが、一般に乳酸菌を培養する液体培地として例えば、Ｍａｎ、Ｒｏｇｏｓａ、Ｓｈａｒｐｅの考案したＭＲＳ培地（メルク社製）、及び牛乳成分を利用したホエー培地、脱脂乳培地等の培地が挙げられる。Lactic acid bacteria may be used alone or in combination of two or more bacteria. Moreover, in the culture | cultivation process mentioned later, the same kind of microbe may be inoculated and cultured in two or more stages, or different bacterial species may be inoculated and cultured in two or more stages.
Lactic acid bacteria may be used after culturing in an agar medium or liquid medium, and then stored by a storage method such as refrigerated storage, frozen storage, dry storage, etc. Using inoculated and cultured (hereinafter abbreviated as seed culture solution) has a fast growth rate of lactic acid bacteria and high activity such as acetaldehyde, diacetyl and other flavors and organic acids. To preferred. The medium used for culturing the seed culture solution is not particularly limited as long as the lactic acid bacteria to be used can grow, but in general, as a liquid medium for culturing lactic acid bacteria, for example, MRS medium (Merck, designed by Man, Rogosa, Sharpe) And whey medium using skim milk ingredients, skim milk medium, and the like.
種培養液を調製するには通常、前記の液体培地に、保存してある乳酸菌を添加し、培養する乳酸菌に適応する好気状態または嫌気状態に維持し、静置または攪拌して、培養すれば良い。 In order to prepare a seed culture solution, usually, a stored lactic acid bacterium is added to the liquid medium, maintained in an aerobic state or an anaerobic state suitable for the lactic acid bacterium to be cultured, and left to stand or stirred for culturing. It ’s fine.
次に、本発明に用いられる乳酸菌培養物の調製方法について説明する。
乳酸菌培養物は、微細藻類と乳酸菌とを含有する培養液中で乳酸菌を培養することにより調製される。ここで、微細藻類と乳酸菌とを含有する培養液は、微細藻類を含有し、乳酸菌を増殖させ得るものであれば特に制限はないが、例えば、水に微細藻類を懸濁した懸濁液に乳酸菌を加える方法、水に微細藻類を加えた湿潤液、ペースト、ケーキ等に乳酸菌を加える方法、上記懸濁液に上記種培養液を加える方法等により得ることができる。
より具体的には、以下に例示する方法により得ることができる。（ｉ）生の藻体や乾燥させた藻体の懸濁液、ペーストに乳酸菌の培養液や乾燥状態の乳酸菌を添加する。（ｉｉ）生の藻体や乾燥させた藻体のケーキに、乳酸菌の培養液を添加する。（ｉｉｉ）乾燥させた藻体に湿潤状態になる量の乳酸菌の培養液を添加する。Next, the preparation method of the lactic acid bacteria culture used for this invention is demonstrated.
The lactic acid bacteria culture is prepared by culturing lactic acid bacteria in a culture solution containing microalgae and lactic acid bacteria. Here, the culture solution containing microalgae and lactic acid bacteria is not particularly limited as long as it contains microalgae and can grow lactic acid bacteria, but for example, in a suspension in which microalgae are suspended in water. It can be obtained by a method of adding lactic acid bacteria, a method of adding lactic acid bacteria to a wet solution obtained by adding microalgae to water, a paste, a cake, or the like, a method of adding the seed culture solution to the suspension.
More specifically, it can be obtained by the method exemplified below. (I) A culture solution of lactic acid bacteria or a dried lactic acid bacterium is added to a raw alga body or a dried suspension or paste of algae. (Ii) A culture solution of lactic acid bacteria is added to a raw alga body or a dried alga body cake. (Iii) A culture solution of lactic acid bacteria in an amount to be in a wet state is added to the dried alga bodies.
これらの中でも、（ｉ）が好ましく、更に（ｉ）において微細藻類として生の藻体の懸濁液を用い、乳酸菌として培養液、特に種培養液を用いるのが、乳酸菌の培養能とフレーバー産生能が高いことから好ましい。
前記藻体の懸濁液やペーストやケーキ、乳酸菌の培養液は、水を含有しているが、微細藻類と乳酸菌とを含有する培養液が、水が足りないときは水を加え、湿潤下や水中に維持する状態にしても良い。水は、滅菌水を用いるのが好ましい。Among these, (i) is preferable, and in (i), a suspension of a raw alga body is used as a microalgae, and a culture solution, particularly a seed culture solution is used as a lactic acid bacterium. It is preferable because of its high performance.
The algae suspension, paste, cake, and lactic acid bacteria culture solution contain water, but if the culture solution containing microalgae and lactic acid bacteria is insufficient, add water and keep it wet. Or you may make it the state maintained underwater. The water is preferably sterilized water.
微細藻類と乳酸菌とを含有する培養液中の微細藻類の含有量は、後述する乳酸菌培養後の収穫工程、乾燥工程で効率を良好にする観点から、乾燥菌体として０．１〜３０質量％が好ましく、１〜２０質量％がより好ましい。
微細藻類と乳酸菌とを含有する培養液中での乳酸菌の培養は、静置培養でも良いし、該培養液が液体であればプロペラ攪拌による攪拌培養でも良い。また、用いる乳酸菌の生育に適するように、培養する系を嫌気状態にしても良いし、好気状態にしても良い。The content of the microalgae in the culture solution containing the microalgae and lactic acid bacteria is 0.1 to 30% by mass as dry cells from the viewpoint of improving efficiency in the harvesting process and the drying process after lactic acid bacteria culture described below. Is preferable, and 1-20 mass% is more preferable.
Cultivation of lactic acid bacteria in a culture solution containing microalgae and lactic acid bacteria may be stationary culture, or may be agitation culture with propeller stirring if the culture solution is liquid. Further, the culture system may be anaerobic or aerobic so as to be suitable for the growth of the lactic acid bacteria used.
乳酸菌の使用量は、乳酸菌が増殖する菌数であれば良いが、夾雑菌の繁殖の抑制を良好にする観点から、乳酸菌の培養を開始する際の乳酸菌数が固形分換算藻類１ｇあたり１０ ５〜１０１１個であるのが好ましく、１０６〜１０１０個であるのがより好ましい。
微細藻類と乳酸菌とを含有する培養液のｐＨは、乳酸菌の培養により生成する乳酸などの酸により培養の過程で変化するが、培養開始時のｐＨが、４．０〜９．０であるのが好ましく、５．０〜７．０がより好ましい。
培養温度は、乳酸菌が増殖可能な温度ならば何れでもよいが、乳酸菌の増殖に好適なこと、微細藻類の有効成分が損なわれないことから、４〜４５℃が好ましく、２０〜４０℃がより好ましい。 The amount of lactic acid bacteria used may be the number of bacteria in which lactic acid bacteria grow, but from the viewpoint of improving the suppression of the propagation of contaminating bacteria, the number of lactic acid bacteria at the start of cultivation of lactic acid bacteria is 10 per gram of algae in terms of solid content. 5-1011Preferably, the number is 106-1010More preferably.
The pH of the culture solution containing microalgae and lactic acid bacteria changes during the cultivation process by an acid such as lactic acid produced by culturing lactic acid bacteria, but the pH at the start of the cultivation is 4.0 to 9.0. Is preferable, and 5.0 to 7.0 is more preferable.
The culture temperature may be any temperature as long as the lactic acid bacteria can grow, but is preferably 4 to 45 ° C. and more preferably 20 to 40 ° C. because it is suitable for the growth of lactic acid bacteria and the active ingredients of microalgae are not impaired. preferable.
培養時間は、乳酸菌を十分増殖させ、優れた血圧低下作用を発揮させるため、例えば、９〜１００時間が好ましく、１５〜７２時間 がより好ましく、１８〜５０時間が特に好ましい。
培養後の乳酸菌数は、優れた血圧低下作用を発揮させ、他の夾雑菌の増加抑制が十分で、微細藻類に特有の味と匂いを良好に減少させるとともに血圧低下作用を発揮させるため、培養開始時の乳酸菌の数の１０〜１０００倍に増加しているのが好ましい。
また、乳酸菌の増殖至適ｐＨを維持するため、培養中に水酸化カリウム、水酸化カルシウム等の塩基性化合物を添加してｐＨを調整してもよいが、乳酸菌が生成する乳酸等により、微細藻類と乳酸菌とを含有する培養液のｐＨが５．０付近まで低下していることが、夾雑菌を低下させるとともに血圧低下作用を発揮させるため、好ましい。The culture time is, for example, preferably 9 to 100 hours, more preferably 15 to 72 hours, and particularly preferably 18 to 50 hours in order to sufficiently proliferate lactic acid bacteria and exert an excellent blood pressure lowering effect.
The number of lactic acid bacteria after culturing exhibits an excellent blood pressure lowering effect, and is sufficient to suppress the increase of other contaminants, to reduce the taste and odor peculiar to microalgae and to exert a blood pressure lowering effect. It is preferable to increase 10 to 1000 times the number of lactic acid bacteria at the start.
In order to maintain the optimum pH for growth of lactic acid bacteria, a basic compound such as potassium hydroxide or calcium hydroxide may be added during the cultivation to adjust the pH. It is preferable that the pH of the culture solution containing algae and lactic acid bacteria is lowered to about 5.0, because it reduces impurities and exerts a blood pressure lowering effect.
本発明においては、微細藻類と乳酸菌とを含有する培養液中に糖を加えておくことが、夾雑菌の生育を抑制することができ、また、微細藻類特有の臭いと味をより減少させることができるので好ましい。この夾雑菌の生育の抑制効果は、微細藻類として夾雑菌が繁殖しやすい乾燥藻体を用いた時に特に顕著である。
前記糖類としては、例えば、単糖類、オリゴ糖類、多糖類等が挙げられる。単糖類としては、例えば、グルコース、ガラクトース、マンノース、フルクトース、リボース、キシロース等が挙げられる。オリゴ糖類としては、例えば、スクロース、マルトース等の二糖類やガラクトオリゴ糖、フラクトオリゴ糖、大豆オリゴ糖、キシロオリゴ糖、ラフィノース等が挙げられる。多糖類としては、例えば、アミロース、アミロペクチン、セルロース、グリコーゲン、β−グルカン、ムコ多糖等が挙げられる。糖類としては、オリゴ糖が好ましく、中でも、ガラクトオリゴ糖、キシロオリゴ糖が好ましい。In the present invention, adding sugar to a culture solution containing microalgae and lactic acid bacteria can suppress the growth of contaminating bacteria, and can further reduce the smell and taste peculiar to microalgae. Is preferable. The effect of suppressing the growth of the contaminating bacteria is particularly remarkable when a dry alga body in which the contaminating bacteria easily propagate is used as a microalgae.
Examples of the saccharide include monosaccharides, oligosaccharides, and polysaccharides. Examples of monosaccharides include glucose, galactose, mannose, fructose, ribose, xylose and the like. Examples of oligosaccharides include disaccharides such as sucrose and maltose, galactooligosaccharides, fructooligosaccharides, soybean oligosaccharides, xylo-oligosaccharides, and raffinose. Examples of the polysaccharide include amylose, amylopectin, cellulose, glycogen, β-glucan, mucopolysaccharide and the like. As the saccharide, oligosaccharide is preferable, and galactooligosaccharide and xylo-oligosaccharide are particularly preferable.
糖類の添加方法には特に制限は無く、例えば、微細藻類と乳酸菌と糖類とを混合しても良いし、予め糖類を添加した微細藻類と乳酸菌とを混合しても良いし、予め糖類を添加した乳酸菌と微細藻類とを混合しても良い。また、糖類は固形のものを使用しても良いが、予め、水等に溶解して水溶液としたものを用いるのが好ましい。
糖類の使用量としては、微細藻類と乳酸菌とを含有する培養液と糖類の合計に対して０．０５〜２０質量％が好ましく、０．１〜１０質量％がより好ましい。There are no particular restrictions on the method of adding saccharides. For example, microalgae, lactic acid bacteria, and saccharides may be mixed, microalgae added with saccharides in advance and lactic acid bacteria may be mixed, or saccharides may be added in advance. The lactic acid bacteria and microalgae may be mixed. Moreover, although a solid saccharide may be used, it is preferable to use a saccharide previously dissolved in water to obtain an aqueous solution.
As the usage-amount of saccharides, 0.05-20 mass% is preferable with respect to the sum total of the culture solution containing microalgae and lactic acid bacteria, and saccharides, and 0.1-10 mass% is more preferable.
かかる乳酸菌培養物の調製過程で生成する乳酸、酢酸等の有機酸類、バクテリオシン類等の抗菌作用により、他の夾雑細菌は減少し、乳酸菌が優越種となる。さらに乳酸菌の生成するアセトアルデヒド、ジアセチル等のフレーバー類により、微細藻類特有の臭いと味が低下し、食品に利用し易い香味となる。 Due to the antibacterial action of organic acids such as lactic acid and acetic acid, bacteriocin and the like produced during the preparation process of such lactic acid bacteria culture, other contaminating bacteria are reduced and lactic acid bacteria become the dominant species. In addition, flavors such as acetaldehyde and diacetyl produced by lactic acid bacteria reduce the smell and taste peculiar to microalgae, resulting in a flavor that is easy to use in foods.
上記で得られた乳酸菌培養物は、そのまま乳酸菌飲料、乳酸菌添加食品として使用することも出来る。
上記で得られた乳酸菌培養物は、培養前の微細藻類の原末とは異なる成分組成を有する特徴がある。
例えば、２４時間培養後の培養物の遊離アミノ酸量に関しては、プロリン、システイン、バリン、ロイシン、イソロイシン、γ−アミノ酪酸（以下、「ＧＡＢＡ」と略記）、リジン、ヒスチジン等が、培養前に比べて、藻体１００ｇあたりの含有量が１０倍以上となり、遊離アミノ酸総量も２倍以上に増加する特徴を有する。
また、分子量１０，０００の分離膜で処理を行った培養前の原末、並びに乳酸菌培養物の含有するタンパク質量を、タンパク質量の一般的な測定法であるＢＣＡ（Bicinchoninic Acid）法により測定した。その結果、例えば、藻体としてスピルリナ原末を用いた場合、分子量１０，０００以下の成分が、２４時間培養後では、培養開始前に比べて約２倍に増加し、高分子量のタンパクが低分子化され、吸収性に優れた成分となっていることが判明した。
また、本発明により得られる乳酸培養物は、従来の本発明に類似する乳酸菌培養物に比較して、高濃度でＧＡＢＡを含有することを特徴とする。例えば、前出の（特許文献３）には、乳類に、Ｌ．ラクチス ＹＩＴ ２０２７を単独で用いた場合、３日間の培養で１５ｍｇ／１００ｍＬであり、Ｌ．カゼイ ＹＩＴ ９０２９と混合培養を行うことにより、３８ｍｇ／１００ｍＬまで増加するとの記載がある。
しかし、本発明による方法によれば、２４時間の培養時間で、１００ｍｇ／１００ｍＬ以上の濃度でのＧＡＢＡの製造が可能となり、ＧＡＢＡ製造法として極めて優れていることが明らかである。The lactic acid bacteria culture obtained above can be used as it is as a lactic acid bacteria beverage or a food supplemented with lactic acid bacteria.
The lactic acid bacteria culture obtained as described above is characterized by having a component composition different from the raw powder of microalgae before culturing.
For example, regarding the amount of free amino acids in the culture after 24 hours of culture, proline, cysteine, valine, leucine, isoleucine, γ-aminobutyric acid (hereinafter abbreviated as “GABA”), lysine, histidine, etc. Thus, the content per 100 g of alga bodies is 10 times or more, and the total free amino acid content is also increased by 2 times or more.
In addition, the amount of protein contained in the bulk powder before culturing treated with the separation membrane having a molecular weight of 10,000 and the lactic acid bacteria culture was measured by the BCA (Bicinchoninic Acid) method which is a general method for measuring the protein amount. . As a result, for example, when spirulina bulk powder is used as an algal body, a component having a molecular weight of 10,000 or less increases about twice as much after the culture for 24 hours as compared to before the start of the culture, and the high molecular weight protein is low. It has been found that it has been molecularized and has an excellent absorbability.
In addition, the lactic acid culture obtained by the present invention is characterized by containing GABA at a higher concentration than the conventional lactic acid bacteria culture similar to the present invention. For example, in the above-mentioned (patent document 3), L. When Lactis YIT 2027 is used alone, it is 15 mg / 100 mL after 3 days of culture. There is a description that it is increased to 38 mg / 100 mL by mixed culture with casei YIT 9029.
However, according to the method of the present invention, it is possible to produce GABA at a concentration of 100 mg / 100 mL or more in a culture time of 24 hours, and it is clear that it is extremely excellent as a GABA production method.
本発明の血圧低下剤は、上記で得られた乳酸菌培養物を有効成分とするものである。本発明の血圧低下剤は、裸錠、フィルムコーティング錠、糖衣錠、腸溶錠、多層錠等の錠剤、顆粒剤、粉末剤、液剤等の形態に調製することができる。これらの形態への調製は、各形態に応じて常法に従い、乳酸菌培養物に懸濁、乾燥、粉砕、成型等を行えばよい。各形態への調製にあたっては、その形態に調製するために一般的に用いられる結着剤、界面活性剤、増粘剤、充填剤、崩壊剤、賦型剤等を用いることができる。また、かかる乳酸菌培養物をそのまま乳酸菌飲料、乳酸菌添加物とすることもできるし、乳酸菌培養物を他の食品に添加することもできる。 The blood pressure lowering agent of the present invention comprises the lactic acid bacteria culture obtained above as an active ingredient. The blood pressure lowering agent of the present invention can be prepared in the form of tablets such as naked tablets, film-coated tablets, sugar-coated tablets, enteric tablets, multilayer tablets, granules, powders, liquids and the like. Preparation into these forms may be carried out by suspending, drying, crushing, molding, etc. in the lactic acid bacteria culture according to each form according to a conventional method. In preparation for each form, binders, surfactants, thickeners, fillers, disintegrants, excipients and the like that are generally used to prepare the form can be used. In addition, the lactic acid bacteria culture can be used as it is as a lactic acid bacteria beverage or lactic acid bacteria additive, or the lactic acid bacteria culture can be added to other foods.
例えば、乾燥処理を行う場合、通常、藻体の水分含有率が４〜７質量％になるように行うが、乳酸菌の菌数が保持できる処理が好ましい。好ましい乾燥方法としては、例えば、凍結乾燥法、噴霧乾燥法等が挙げられるが、経済的であることから、噴霧乾燥法がより好ましい。乾燥処理する際の乾燥温度は、排風温度が高温な程、生産効率は上がるが、微細藻類の品質が良好で、乳酸菌数も低下しないことから、品温が３０〜７０℃となる範囲で乾燥処理するのが好ましく、より好ましくは４０〜６０℃となる範囲である。尚、本発明において品温とは、乾燥後の試料温度をいうものとする。 For example, when performing a drying process, it is normally performed so that the moisture content of the algal cells is 4 to 7% by mass, but a process capable of maintaining the number of lactic acid bacteria is preferable. Preferable drying methods include, for example, a freeze-drying method and a spray-drying method, but the spray-drying method is more preferable because it is economical. The drying temperature at the time of drying treatment increases the production efficiency as the exhaust air temperature is higher, but the quality of microalgae is good and the number of lactic acid bacteria does not decrease, so the product temperature is in the range of 30 to 70 ° C. It is preferable to dry-process, More preferably, it is the range used as 40-60 degreeC. In the present invention, the product temperature refers to the sample temperature after drying.
錠剤への調製は、例えば、上記で得られた粉末を、公知の打錠法によって打錠すればよい。
液剤にする場合は、例えば上記で得られた粉末を水に分散させてもよいし、培養の終了した乳酸菌培養物を、そのままあるいは水等で希釈して調製することができる。Preparation into a tablet may be performed by, for example, tableting the powder obtained above by a known tableting method.
In the case of preparing a solution, for example, the powder obtained above may be dispersed in water, or the cultured lactic acid bacteria culture can be prepared as it is or diluted with water or the like.
本発明の血圧低下剤の患者への摂取量は、患者の性別、年齢、症状等を考慮して決定することが好ましいが、一般的に、乳酸菌培養物換算で、０．２〜１０ｇ/日、特に０．５〜８ｇ/日が好ましい。これらは、１度にまとめて摂取してもよいし、１日摂取量を数度に分けて摂取してもよい。 The intake of the blood pressure lowering agent of the present invention to the patient is preferably determined in consideration of the sex, age, symptoms, etc. of the patient, but generally 0.2 to 10 g / day in terms of lactic acid bacteria culture. In particular, 0.5 to 8 g / day is preferable. These may be ingested all at once, or the daily intake may be divided into several degrees.
次に、本発明の血圧低下剤の血圧低下作用について説明する。
本発明者らは、上記で得られた乳酸菌培養物に関して、種々の生活習慣病の主要因と考えられている高血圧症に対する効果の確認を、実験動物を用いて行った。
即ち、高血圧自然発症ラット（ＳＨＲ系）に、スピルリナ乳酸菌培養物、及びクロレラ乳酸菌培養物を固形飼料に混餌させ、自由摂取を行った後に、血圧測定を行い、血圧低下作用を確認した。実験群は、対象群として固形飼料を与えた群を設定し、その他に、被験物質を混餌した群として、スピルリナ原末（被験物質１）を固形飼料に混餌した群、スピルリナ乳酸菌培養物（被験物質２）を固形飼料に混餌した群、クロレラ原末（被験物質３）を固形飼料に混餌した群、クロレラ乳酸菌培養物（被験物質４）を固形飼料に混餌した群を設定した。その結果、スピルリナ乳酸菌培養物又はクロレラ乳酸菌培養物を混餌した群は、何れも、対照群、スピルリナ原末、及びクロレラ原末を混餌した群に比較し、強い血圧低下作用を有することが明らかとなった（試験例参照）。Next, the blood pressure lowering action of the blood pressure lowering agent of the present invention will be described.
The inventors of the present invention confirmed the effect on hypertension, which is considered to be the main cause of various lifestyle-related diseases, using experimental animals with respect to the lactic acid bacteria culture obtained above.
In other words, spontaneously hypertensive rats (SHR strain) were mixed with spirulina lactic acid bacteria culture and chlorella lactic acid bacteria culture in solid feed, and after free intake, blood pressure was measured to confirm blood pressure lowering action. In the experimental group, a group fed with solid feed was set as a target group, and in addition, as a group fed with a test substance, a group of spirulina powder (test substance 1) mixed with a solid feed, a spirulina lactic acid bacteria culture (test) A group in which substance 2) was mixed with solid feed, a group in which chlorella bulk powder (test substance 3) was mixed in solid feed, and a group in which chlorella lactic acid bacteria culture (test substance 4) was mixed in solid feed were set. As a result, it is clear that the group fed with the spirulina lactic acid bacteria culture or the chlorella lactic acid bacteria culture has a strong blood pressure lowering effect as compared with the control group, the spirulina bulk powder, and the group fed with the chlorella bulk powder. (See test example).
なお、乳酸菌培養物の安全性確認のため、マウスによる急性毒性を調べた。マウスは、生後５週令のｄｄＹ−Ｎ系マウス（体重２０〜２６ｇ）を雄、雌各１０頭使用した。投与方法は、培養物を微粉砕し、ＣＭＣ１％溶液に懸濁して胃ゾンデによる投与可能最高濃度１０質量％懸濁液を１回、経口強制投与した。培養物を投与後、１週間観察した。雄、雌ともＬＤ５０は、６，０００ｍｇ／ｋｇ以上であり、食品としての安全性が確認された。In order to confirm the safety of lactic acid bacteria cultures, the acute toxicity by mice was examined. As the mice, 10 male and 10 female ddY-N mice (body weight 20 to 26 g) 5 weeks old were used. In the administration method, the culture was finely pulverized, suspended in a CMC 1% solution, and a 10% by mass suspension capable of being administered with a gastric sonde was orally gavaged once. The culture was observed for 1 week after administration. LD 50 was 6,000 mg / kg or more for both males and females, confirming food safety.
次に実施例を示して本発明をさらに詳細に説明するが、本発明は以下の実施例に限定されるものではない。 EXAMPLES Next, although an Example is shown and this invention is demonstrated further in detail, this invention is not limited to a following example.
（実施例１） スピルリナを用いた乳酸菌培養物の調製（１）と乳酸菌数の測定
２５００Ｌ容量培養槽にガラクトオリゴ糖２５ｋｇ、水道水８２５ｋｇを仕込み、加熱殺菌後、スピルリナ原末１００ｋｇを仕込み、乳酸菌ラクトバチルス ブレビスの種培養液５０ｋｇを接種した。３７℃、７２時間、通気プロペラ撹拌して乳酸菌を培養した。培養時間０、９、１２、１５、１８、２４、４８、及び７２時間に培養液をサンプリングし、乳酸菌数を測定した。Example 1 Preparation of Lactic Acid Bacteria Culture Using Spirulina (1) and Measurement of Lactic Acid Bacteria 25 kg of galactooligosaccharide and 825 kg of tap water are charged in a 2500 L culture tank, and after heat sterilization, 100 kg of spirulina bulk powder is charged. 50 kg of Bacillus brevis seed culture was inoculated. Lactic acid bacteria were cultured with aeration propeller stirring at 37 ° C. for 72 hours. The culture solution was sampled at culture times of 0, 9, 12, 15, 18, 24, 48, and 72 hours, and the number of lactic acid bacteria was measured.
なお、乳酸菌数測定は、以下の手順で行った。乳酸菌培養液１．０ｍｌをリン酸緩衝生理食塩水１９ｍｌに懸濁し懸濁液を調整した。懸濁液をさらにリン酸緩衝生理食塩水により、１０倍、１０２倍、１０３倍、１０４倍、１０５倍、１０６倍、及び１０７倍になるように希釈して試料希釈液を得た。１０００ｍｌのＭＲＳ培地（メルク社製 Ｃａｔ.No.10661）に１５ｇの寒天を添加して作製したＭＲＳ寒天培地に、０．１ｍｌの希釈液を塗沫し、３５℃、４８時間培養した。乳酸菌のコロニーが３０〜３００個確認できた寒天培地を用い、該寒天培地のコロニー数を測定し、この数に希釈倍率を掛けて得られた数を乳酸菌のコロニー数とした。The number of lactic acid bacteria was measured according to the following procedure. 1.0 ml of lactic acid bacteria culture solution was suspended in 19 ml of phosphate buffered saline to prepare a suspension. The suspension further with phosphate-buffered saline, 10x, 10 twice, 10 three times, 10 4 times, 10 5 times, 10 6 times, and sample diluent was diluted to 10 7 times Got. A 0.1 ml dilution was smeared on an MRS agar medium prepared by adding 15 g of agar to 1000 ml of MRS medium (Cat. No. 10661 manufactured by Merck & Co., Inc.) and cultured at 35 ° C. for 48 hours. Using an agar medium in which 30 to 300 colonies of lactic acid bacteria were confirmed, the number of colonies in the agar medium was measured, and the number obtained by multiplying this number by the dilution factor was taken as the number of colonies of lactic acid bacteria.
（アミノ酸量の測定）
サンプリングした各培養液の一部を、噴霧乾燥してアミノ酸分析に供した。スピルリナの乳酸菌培養物各２００ ｍｇを５ｍＬの水に添加後、懸濁させ、３０分間放置した。遠心分離機（３０００ｒｐｍ、１０分間）により、上清と不溶物に分離し、上清１ｍＬを採取した。これに、１０％（ｗ/ｖ）トリクロロ酢酸溶液１ｍＬを添加し、遠心分離を行った。得られた上清について、o-フタルアルデヒド法（ポストカラム検出）により、ＨＰＬＣ分析を行い、アミノ酸量を測定した。
（ＨＰＬＣ条件）
カラム：日立＃４６１９（４ｍｍ×１５ｃｍ）
温度：６０℃（ポストカラム反応時も同温度）
流速：溶離液；０.４ｍＬ/ｍｉｎ. 反応液；０.５ｍＬ/ｍｉｎ.
検出：蛍光検出器
各培養時間における培養物中のＬ−グルタミン酸量、ＧＡＢＡ量、乳酸菌数を表１に示す。(Measurement of amino acid content)
A part of each sampled culture solution was spray-dried and subjected to amino acid analysis. Each 200 mg of Spirulina lactic acid bacteria culture was added to 5 mL of water, suspended, and allowed to stand for 30 minutes. The supernatant and insoluble material were separated by a centrifuge (3000 rpm, 10 minutes), and 1 mL of the supernatant was collected. To this, 1 mL of a 10% (w / v) trichloroacetic acid solution was added and centrifuged. The obtained supernatant was subjected to HPLC analysis by o-phthalaldehyde method (post column detection) to measure the amount of amino acids.
(HPLC conditions)
Column: Hitachi # 4619 (4 mm x 15 cm)
Temperature: 60 ° C (same temperature during post-column reaction)
Flow rate: Eluent: 0.4 mL / min. Reaction solution: 0.5 mL / min.
Detection: Fluorescence detector Table 1 shows the amount of L-glutamic acid, the amount of GABA, and the number of lactic acid bacteria in the culture at each culture time.
（実施例２） スピルリナを用いた乳酸菌培養物の調製（２）
本実施例では、５０％（ｗ/ｗ）乳酸溶液でｐＨを５．０に、培養温度を３０℃に保持した以外は、実施例１と同様にして、乳酸菌を培養した。培養時間０、９、１２、１５、１８、２１、２４時間に培養液をサンプリングし、ＧＡＢＡ量（ｍｇ／１００ｇ）を測定した。
（実施例３） スピルリナを用いた乳酸菌培養物の調製（３）
本実施例では、ガラクトオリゴ糖の換わりにキシロオリゴ糖を用いる他は、実施例２と同様にして、ＧＡＢＡ量（ｍｇ／１００ｇ）を測定した。
培養時間２４時間までの実施例１と実施例２のＧＡＢＡ量（ｍｇ/１００ｇ）の比較を図１に、実施例２と実施例３の結果を表２に示す。
実施例１〜３より、ｐＨ及び培養温度を適宜選択することにより、短時間でより多くのＧＡＢＡ産生が可能となった。
(Example 2) Preparation of lactic acid bacteria culture using Spirulina (2)
In this example, lactic acid bacteria were cultured in the same manner as in Example 1 except that the pH was adjusted to 5.0 and the culture temperature was maintained at 30 ° C. with a 50% (w / w) lactic acid solution. The culture solution was sampled at culture times of 0, 9, 12, 15, 18, 21, and 24 hours, and the amount of GABA (mg / 100 g) was measured.
(Example 3) Preparation of lactic acid bacteria culture using Spirulina (3)
In this example, the amount of GABA (mg / 100 g) was measured in the same manner as in Example 2 except that xylo-oligosaccharide was used instead of galactooligosaccharide.
A comparison of GABA amounts (mg / 100 g) between Example 1 and Example 2 up to a culture time of 24 hours is shown in FIG. 1, and the results of Example 2 and Example 3 are shown in Table 2.
From Examples 1 to 3, it was possible to produce more GABA in a short time by appropriately selecting pH and culture temperature.
（実施例４） クロレラを用いた乳酸菌培養物の調製と乳酸菌数の測定
実施例１において、スピルリナの代わりにクロレラを用いた以外は、実施例１と同様にして乳酸菌培養物を調製し、乳酸菌数、アミノ酸量を測定した。各培養時間における培養物中のＬ−グルタミン酸量、ＧＡＢＡ量、乳酸菌数を表３に示す。(Example 4) Preparation of lactic acid bacteria culture using chlorella and measurement of the number of lactic acid bacteria In Example 1, a lactic acid bacteria culture was prepared in the same manner as in Example 1 except that chlorella was used instead of Spirulina. The number and the amount of amino acids were measured. Table 3 shows the amount of L-glutamic acid, the amount of GABA, and the number of lactic acid bacteria in the culture at each culture time.
（試験例） 本発明の血圧低下剤による血圧低下作用の確認
本発明の血圧低下剤の血圧低下作用を明らかにするため、以下の実験動物を用いた試験を行った。
（試験物質）
対象群：固形飼料ＭＦ(オリエンタル酵母株式会社製)
被験物質１：スピルリナ原末
被験物質２：乳酸発酵スピルリナ（実施例１で得られた乳酸菌培養物）
被験物質３：クロレラ原末
被験物質４：乳酸発酵クロレラ（実施例４で得られた乳酸菌培養物）
（被験物質の調整及び投与）
調整方法：固形飼料ＭＦに被験物質１〜２をそれぞれ１５％混合した。
投与経路：経口
投与方法：自由摂取
投与容量：給餌量の１５％
投与期間：８週間
試験動物
動物種、系統：ラット、ＳＨＲ（１群８匹）
性別、搬入週齢：雄、６週齢
馴化期間：搬入後約１週間馴化を兼ねて飼育し、その期間の一般状態を観察した。
（血圧測定法）
血圧の測定は尾動脈の血圧を非観血的に測定するＴａｉｌ-ｃｕｆｆ法により行った。測定方法は、試験物質の投与前（７週目）、投与後１週間毎の計８ポイントで実施した。血圧の各ポイントでの測定は３回ずつ実施し、その平均値を測定値とした。なお、試験実施に先立ち供試動物の最高血圧の平均値が各動物とも平均１６０ｍｍＨｇ以上であることを確認し、試験を行った。
結果を表４に示す。(Test example) Confirmation of blood pressure lowering action of the blood pressure lowering agent of the present invention In order to clarify the blood pressure lowering action of the blood pressure lowering agent of the present invention, a test using the following experimental animals was performed.
(Test substance)
Target group: Solid feed MF (manufactured by Oriental Yeast Co., Ltd.)
Test substance 1: Spirulina bulk powder Test substance 2: Lactic acid fermented spirulina (Lactic acid bacteria culture obtained in Example 1)
Test substance 3: Chlorella bulk powder Test substance 4: Lactic acid fermentation chlorella (Lactic acid bacteria culture obtained in Example 4)
(Test substance adjustment and administration)
Adjustment method: 15% of test substances 1 and 2 were mixed with solid feed MF.
Administration route: Oral Administration method: Free intake Administration volume: 15% of food consumption
Administration period: 8 weeks Test animal Animal species, strain: rat, SHR (8 per group)
Sex, age of import: male, 6 weeks of age Acclimatization period: Breeding for about one week after accompaniment, and observing the general condition during that period.
(Blood pressure measurement method)
The blood pressure was measured by the Tail-cuff method for noninvasively measuring the blood pressure of the tail artery. The measurement was performed at a total of 8 points before administration of the test substance (7 weeks) and every week after administration. The measurement at each point of blood pressure was performed three times, and the average value was taken as the measurement value. Prior to the test, it was confirmed that the average value of the maximum blood pressure of the test animals was 160 mmHg or more on average for each animal.
The results are shown in Table 4.
表５及び図２には、０週間目の血圧を基準としたＸ週目の血圧上昇率を以下の式にて評価した結果を示す。
血圧上昇率（％）＝（Ｘ週目の血圧−０週目の血圧）/（０週目の血圧）×１００（％）
Blood pressure increase rate (%) = (blood pressure at week X−blood pressure at week 0) / (blood pressure at week 0) × 100 (%)
本試験例より明らかなように、スピルリナ乳酸菌培養物及びクロレラ乳酸菌培養物は、各々培養前のスピルリナ原末及びクロレラ原末に比較して、血圧上昇抑制効果が確認された。
例えば、６週間後においては、対象群では、約３０％の血圧上昇が見られるのに対して、スピルリナ及びクロレラでは、その上昇率が２０〜２５％に抑えられ、更には、本発明のスピルリナ及びクロレラの乳酸発酵品においては、１２〜１５％程度に抑制されることが明らかとなり、本発明に係る乳酸発酵品の血圧低下作用が確認された。As is clear from this test example, the spirulina lactic acid bacteria culture and the chlorella lactic acid bacteria culture were confirmed to have an effect of suppressing blood pressure increase as compared with the spirulina bulk powder and the chlorella bulk powder before culture, respectively.
For example, after 6 weeks, the blood pressure increase of about 30% is observed in the subject group, whereas in spirulina and chlorella, the increase rate is suppressed to 20 to 25%. Furthermore, the spirulina of the present invention is further reduced. In addition, the lactic acid fermented product of Chlorella was found to be suppressed to about 12 to 15%, and the blood pressure lowering action of the lactic acid fermented product according to the present invention was confirmed.
本発明は、医薬品産業、健康食品、食品等の分野で利用が可能である。 The present invention can be used in fields such as the pharmaceutical industry, health food, and food.
Claims (16)
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| JP2007526843A JP4762987B2 (en) | 2005-04-15 | 2006-04-13 | Antihypertensive agent obtained by lactic acid bacteria culture |
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| JP2007526843A JP4762987B2 (en) | 2005-04-15 | 2006-04-13 | Antihypertensive agent obtained by lactic acid bacteria culture |
| PCT/JP2006/307856 WO2006112364A1 (en) | 2005-04-15 | 2006-04-13 | Hypotensive agent produced by cultivation of lactic acid bacterium |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2007526843A Active JP4762987B2 (en) | 2005-04-15 | 2006-04-13 | Antihypertensive agent obtained by lactic acid bacteria culture |
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| Country | Link |
|---|---|
| JP (1) | JP4762987B2 (en) |
| KR (1) | KR20080004515A (en) |
| CN (1) | CN101160136A (en) |
| WO (1) | WO2006112364A1 (en) |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2933848B1 (en) * | 2008-07-18 | 2019-08-09 | Roquette Freres | COMPOSITION OF SOLUBLE INDIGESTIBLE FIBERS AND EUKARYOTIC ORGANISMS HAVING POLYSACCHARIDE WAVES USED IN THE WELL-BEING FIELD |
| FR2935228B1 (en) * | 2009-09-30 | 2013-02-08 | Roquette Freres | COMPOSITION OF BRANCHED MALTODEXTRINS AND EUKARYOTIC ORGANISMS HAVING POLYSACCHARIDE WALLS USED IN THE WELL-BEING FIELD |
| CN102763809A (en) * | 2012-07-05 | 2012-11-07 | 昆明豪原特自控有限公司 | Biological feed additive and preparation method of biological feed additive |
| WO2017216818A1 (en) * | 2016-06-17 | 2017-12-21 | Ocean Farma Srl | Anaerobic growth of bacteria in unicellular alga |
| IT201800005108A1 (en) | 2018-05-07 | 2019-11-07 | PROCEDURE FOR THE SYNTHESIS OF SILVER NANOPARTICLES BY MEANS OF PROBIOTIC STRAINS GROWN IN ALGAE, NANOPARTICLES SO OBTAINED AND RELATED USES | |
| KR102287187B1 (en) * | 2018-12-31 | 2021-08-06 | 재단법인 전남바이오산업진흥원 | Prebiotics composition comprising extract of Pyropia yezoensis as effective component and uses thereof |
Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5538336A (en) * | 1978-09-12 | 1980-03-17 | Kurorera Kogyo Kk | Extraction of hypotensive substance |
| JPS6245532A (en) * | 1985-08-23 | 1987-02-27 | Nisshin Oil Mills Ltd:The | Medicine and health supplementary food having inhibitory effect on blood pressure rise |
| JPS63139127A (en) * | 1986-11-29 | 1988-06-10 | Michiko Kamijo | Spreading plaster for athlete' foot |
| JPS63157963A (en) * | 1986-12-22 | 1988-06-30 | Toshiyuki Ota | Production of chlorella food |
| JPH0751057A (en) * | 1993-08-11 | 1995-02-28 | Kawasaki Steel Corp | Material containing proliferation promoting substance for lactic acid bacteria and bifidus bacteria |
| JP2000102364A (en) * | 1998-09-29 | 2000-04-11 | Minoru Yoneda | Spirulina function-developing food |
| JP2002241305A (en) * | 2001-02-14 | 2002-08-28 | Yakult Honsha Co Ltd | Prophylactic/therapeutic agent for hypertension |
| JP2003088339A (en) * | 2001-09-19 | 2003-03-25 | Kentekku:Kk | Method for producing super chlorella |
| JP2004081206A (en) * | 2002-06-26 | 2004-03-18 | Dainippon Ink & Chem Inc | Method for processing spirulina |
| JP2004357611A (en) * | 2003-06-05 | 2004-12-24 | Masayasu Onishi | Medium comprising solubilized liquid of bean curd refuse, method for producing the same, method for culturing microorganism by using the medium, method for producing useful material, and food material |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0647551B2 (en) * | 1986-12-01 | 1994-06-22 | 株式会社ヤクルト本社 | Antihypertensive agent |
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2006
- 2006-04-13 WO PCT/JP2006/307856 patent/WO2006112364A1/en active Application Filing
- 2006-04-13 CN CNA2006800120134A patent/CN101160136A/en active Pending
- 2006-04-13 JP JP2007526843A patent/JP4762987B2/en active Active
- 2006-04-13 KR KR1020077024509A patent/KR20080004515A/en not_active Application Discontinuation
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5538336A (en) * | 1978-09-12 | 1980-03-17 | Kurorera Kogyo Kk | Extraction of hypotensive substance |
| JPS6245532A (en) * | 1985-08-23 | 1987-02-27 | Nisshin Oil Mills Ltd:The | Medicine and health supplementary food having inhibitory effect on blood pressure rise |
| JPS63139127A (en) * | 1986-11-29 | 1988-06-10 | Michiko Kamijo | Spreading plaster for athlete' foot |
| JPS63157963A (en) * | 1986-12-22 | 1988-06-30 | Toshiyuki Ota | Production of chlorella food |
| JPH0751057A (en) * | 1993-08-11 | 1995-02-28 | Kawasaki Steel Corp | Material containing proliferation promoting substance for lactic acid bacteria and bifidus bacteria |
| JP2000102364A (en) * | 1998-09-29 | 2000-04-11 | Minoru Yoneda | Spirulina function-developing food |
| JP2002241305A (en) * | 2001-02-14 | 2002-08-28 | Yakult Honsha Co Ltd | Prophylactic/therapeutic agent for hypertension |
| JP2003088339A (en) * | 2001-09-19 | 2003-03-25 | Kentekku:Kk | Method for producing super chlorella |
| JP2004081206A (en) * | 2002-06-26 | 2004-03-18 | Dainippon Ink & Chem Inc | Method for processing spirulina |
| JP2004357611A (en) * | 2003-06-05 | 2004-12-24 | Masayasu Onishi | Medium comprising solubilized liquid of bean curd refuse, method for producing the same, method for culturing microorganism by using the medium, method for producing useful material, and food material |
Also Published As
| Publication number | Publication date |
|---|---|
| KR20080004515A (en) | 2008-01-09 |
| CN101160136A (en) | 2008-04-09 |
| WO2006112364A1 (en) | 2006-10-26 |
| JPWO2006112364A1 (en) | 2008-12-11 |
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