JP2019170250A - Products fermented with lactic acid bacteria - Google Patents
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本発明は、乳酸菌発酵物に関する。より詳しくは、本発明は、乳酸菌発酵物、該発酵物を含有する組成物及び該発酵物の製造方法に関する。 The present invention relates to a fermented lactic acid bacterium. More specifically, the present invention relates to a lactic acid bacteria fermented product, a composition containing the fermented product, and a method for producing the fermented product.
オタネニンジン（Ｐａｎａｘ ｇｉｎｓｅｎｇ， Ｃ．Ａ． Ｍｅｙｅｒ）は中国、朝鮮半島、日本を中心に古来より生薬として利用されてきた。その有用成分はジンセノシド（ｇｉｎｓｅｎｏｓｉｄｅ）と呼ばれるダマラン骨格を有したサポニン類（ニンジンサポニン）と言われ、抗酸化や、抗糖尿病、抗肥満、高血圧、抗がんなど様々な生理作用が研究により見出されている（非特許文献１）。 Panax ginseng (CA Meyer) has been used as a crude drug since ancient times mainly in China, the Korean peninsula and Japan. Its useful ingredients are called saponins (carrot saponins) with a damaran skeleton called ginsenoside, and various physiological effects such as antioxidant, anti-diabetic, anti-obesity, hypertension, and anti-cancer have been found by research. (Non-Patent Document 1).
ジンセノシドＲｂ１、Ｒｂ２、Ｒｃ、Ｒｄのようなプロトパナキサジオール型のニンジンサポニンは腸内細菌によって、様々な代謝物に代謝変換された後に腸管壁を通過して血中へ移行するとされ、２０Ｓ−ｐｒｏｔｏｐａｎａｘａｄｉｏｌ ２０−Ｏ−ｂｅｔａ−Ｄ−ｇｌｕｃｏｐｙｒａｎｏｓｉｄｅ（別名：Ｍ１、ｃｏｍｐｏｕｎｄ Ｋ）がこれまでニンジンサポニンによる生理作用の中心的な活性成分と考えられている。 Protopanaxadiol-type carrot saponins such as ginsenosides Rb1, Rb2, Rc, Rd are metabolized by intestinal bacteria into various metabolites and then translocated into the blood through the intestinal wall. Protopanaxadiol 20-O-beta-D-glucopyranoside (also known as M1, compound K) has been considered as a central active component of the physiological action by carrot saponin.
一方で、「Ｍ１」への代謝の一つ前の産物である「ジンセノシドＦ２」と呼ばれる成分には「Ｍ１」と同様に抗腫瘍活性や抗肥満効果だけでなく（非特許文献２、３）、インスリン抵抗性を緩和する効果（特許文献１）や育毛効果についても報告されており（非特許文献４）、血中へもマウス実験により「ジンセノシドＦ２」の形のまま移行が報告されている（非特許文献５）ことからも「Ｍ１」とは異なる作用機序並びに健康維持に関する生理活性を発揮する可能性も期待されている。 On the other hand, the component called “ginsenoside F2”, which is the product immediately before the metabolism to “M1”, has not only antitumor activity and anti-obesity effect like “M1” (Non-patent Documents 2 and 3). In addition, the effect of alleviating insulin resistance (Patent Document 1) and the hair-growth effect have also been reported (Non-Patent Document 4), and migration into the blood in the form of “ginsenoside F2” has been reported by mouse experiments. (Non-Patent Document 5) From this, the mechanism of action different from “M1” and the possibility of exerting physiological activity relating to health maintenance are also expected.
乳酸菌によりジンセノシドに働きかけ、生理活性上中心的な代謝物である「Ｍ１」を高含有する発酵オタネニンジンについての先行技術文献はいくつか報告されている。しかしながら代謝において「Ｍ１」のひとつ前の段階の成分である「ジンセノシドＦ２」までの代謝を示し、高含有化に繋がる製造方法は代謝の制御上困難であった。これは一般的に乳酸菌を含め微生物は株が異なれば代謝活性の有無、その強さも異なり、望まれた活性を有する特定の株の取得が必要となり多数の検体を評価しなければならない事にも起因する。 Several prior art documents have been reported on fermented ginseng which acts on ginsenoside by lactic acid bacteria and contains a high content of "M1" which is a central metabolite in terms of physiological activity. However, in the metabolism, the process up to “ginsenoside F2”, which is a component in the previous stage of “M1”, is shown, and the production method leading to high content is difficult in terms of metabolic control. This is because, in general, microorganisms including lactic acid bacteria have different metabolic activity if their strains differ, and it is necessary to acquire a specific strain having the desired activity, and many samples must be evaluated. to cause.
本発明は、新規発酵物、該発酵物を含有する組成物及び該発酵物の製造方法等に関するものである。 The present invention relates to a novel fermented product, a composition containing the fermented product, a method for producing the fermented product, and the like.
本発明者らは、前記課題を解決せんと鋭意検討した結果、一般的に豊富に含まれる「ジンセノシドＲｂ１」から「Ｍ１」への代謝の過程で、「ジンセノシドＦ２」の段階で代謝を停止・もしくは抑制しうる乳酸菌株を独自に自然界から入手する事に成功し、さらなる検討を重ねて本発明を完成するに至った。 As a result of diligent studies to solve the above problems, the present inventors stopped the metabolism at the stage of “ginsenoside F2” in the process of metabolism from “ginsenoside Rb1”, which is generally abundantly contained, to “M1”. Alternatively, the present inventors have succeeded in obtaining a lactic acid strain that can be suppressed from nature, and have completed further studies and completed the present invention.
即ち、本発明は、下記〔１〕〜〔１１〕に関する。
〔１〕 オタネニンジン（高麗人参；Ｋｏｒｅａｎ ｇｉｎｓｅｎｇ：Ｐａｎａｘ Ｃ．Ａ．Ｍｅｙｅｒ）、三七ニンジン（Ｐａｎａｘ ｎｏｔｏｇｉｎｓｅｎｇ Ｂｕｒｋ．）、アメリカニンジン（Ｐａｎａｘ ｑｕｉｎｑｕｅｆｏｌｉｕｍ Ｌ．）、竹節ニンジン（Ｐａｎａｘ ｊａｐｏｎｉｃｕｓ Ｃ．Ａ．Ｍｅｙｅｒ）、ヒマラヤニンジン（Ｐａｎａｘ Ｐｓｅｕｄｏ−ｇｉｎｓｅｎｇ Ｑａｌｌ．Ｓｕｂｓｐ．Ｈｉｍａｌａｉｃｕｓ Ｈａｒａ）及びベトナムニンジン（Ｐａｎａｘ Ｖｕｅｔｎａｍｅｎｓｉｓ Ｈａ ｅｔ Ｇｒｕｓｈｖ．）からなる群より選択される少なくとも１種のウコギ科薬用ニンジンの、ラクトバチルス・パラカゼイ ＤＮＢＬ１８８７株（受託番号ＮＩＴＥ Ｐ−０２６６２）及び／又はラクトバチルス・パラカゼイ ＤＮＢＬ１８８８株（受託番号ＮＩＴＥ Ｐ−０２６６３）による発酵物。
〔２〕 発酵物がジンセノシドＦ２（Ｆ２）を含有してなる、前記〔１〕記載の発酵物。
〔３〕 ジンセノシドＦ２（Ｆ２）とジンセノシドＲｅ（Ｒｅ）の含有量比（Ｆ２／Ｒｅ）が０．００１以上である、発酵物。
〔４〕 前記〔１〕又は〔２〕記載の発酵物である、前記〔３〕記載の発酵物。
〔５〕 前記〔１〕〜〔４〕のいずれかに記載の発酵物を含有してなる、組成物。
〔６〕 抗腫瘍、抗肥満、育毛、抗健忘症、肝保護作用、抗動脈硬化、老化防止、抗高脂血症、抗血栓、血圧降下作用、免疫機能改善、血糖値低下作用、鎮痛作用、強心作用、抗炎症作用、末梢血流改善作用、止血作用、抗酸化作用、抗筋委縮作用、骨量増加作用及び抗ストレス用から選択された少なくとも１種の用途に使用するための、前記〔１〕〜〔５〕のいずれかに記載の発酵物又は組成物。
〔７〕 オタネニンジン（高麗人参；Ｋｏｒｅａｎ ｇｉｎｓｅｎｇ：Ｐａｎａｘ Ｃ．Ａ．Ｍｅｙｅｒ）、三七ニンジン（Ｐａｎａｘ ｎｏｔｏｇｉｎｓｅｎｇ Ｂｕｒｋ．）、アメリカニンジン（Ｐａｎａｘ ｑｕｉｎｑｕｅｆｏｌｉｕｍ Ｌ．）、竹節ニンジン（Ｐａｎａｘ ｊａｐｏｎｉｃｕｓ Ｃ．Ａ．Ｍｅｙｅｒ）、ヒマラヤニンジン（Ｐａｎａｘ Ｐｓｅｕｄｏ−ｇｉｎｓｅｎｇ Ｑａｌｌ．Ｓｕｂｓｐ．Ｈｉｍａｌａｉｃｕｓ Ｈａｒａ）及びベトナムニンジン（Ｐａｎａｘ Ｖｕｅｔｎａｍｅｎｓｉｓ Ｈａ ｅｔ Ｇｒｕｓｈｖ．）からなる群より選択される少なくとも１種のウコギ科薬用ニンジンを、ラクトバチルス・パラカゼイ ＤＮＢＬ１８８７株（受託番号ＮＩＴＥ Ｐ−０２６６２）及び／又はラクトバチルス・パラカゼイ ＤＮＢＬ１８８８株（受託番号ＮＩＴＥ Ｐ−０２６６３）により発酵させることを特徴とする、発酵物の製造方法。
〔８〕 発酵物がジンセノシドＦ２（Ｆ２）を含有してなる、前記〔７〕記載の方法。
〔９〕 発酵物における、ジンセノシドＦ２（Ｆ２）とジンセノシドＲｅ（Ｒｅ）の含有量比（Ｆ２／Ｒｅ）が０．００１以上である、前記〔７〕又は〔８〕記載の方法。
〔１０〕 ラクトバチルス・パラカゼイ ＤＮＢＬ１８８７株（受託番号ＮＩＴＥ Ｐ−０２６６２）。
〔１１〕 ラクトバチルス・パラカゼイ ＤＮＢＬ１８８８株（受託番号ＮＩＴＥ Ｐ−０２６６３）。
That is, the present invention relates to the following [1] to [11].
[1] Ginseng (Korean ginseng: Panax CA Meyer), Panax notoginseng Burk., American carrot (Panax quinquefolium L.), Bamboo carrot (Pane. BLAT Lactobacillus N, which is at least one species of Saccharomyces cerevisiae N, selected from the group consisting of Panax Pseudo-ginseng Qall. Subsp. Number NITE P-02662) and / or Lactobacillus paracasei DNBL1 Fermented product from 888 strain (Accession Number NITE P-02663).
[2] The fermented product according to the above [1], wherein the fermented product contains ginsenoside F2 (F2).
[3] A fermented product having a content ratio (F2 / Re) of ginsenoside F2 (F2) and ginsenoside Re (Re) of 0.001 or more.
[4] The fermented product according to [3], which is the fermented product according to [1] or [2].
[5] A composition comprising the fermented product according to any one of [1] to [4].
[6] Anti-tumor, anti-obesity, hair growth, anti-amnesia, hepatoprotective action, anti-arteriosclerosis, anti-aging, anti-hyperlipidemia, anti-thrombosis, blood pressure lowering action, immune function improvement, blood sugar level lowering action, analgesic action For use in at least one application selected from cardiotonic action, anti-inflammatory action, peripheral blood flow improving action, hemostasis action, antioxidant action, antimuscular atrophy action, bone mass increasing action and anti-stress use, The fermented product or composition according to any one of [1] to [5].
[7] Ginseng (Korean Ginseng: Panax CA Meyer), Panax notoginseng Burk., Panax quinquefolium L., Bamboo Carrot (Pane. At least one kind of carrot for the genus Saccharidae selected from the group consisting of Panax Pseudo-ginseng Qall. Subsp. Himalaicus Hara and Vietnamese carrot (Panax Vuetnamensis Ha et Grushv.) Number NITE P-02662) and / or Lactobacillus paracasei DNBL1 A method for producing a fermented product, characterized by fermenting with 888 strain (Accession No. NITE P-02663).
[8] The method according to [7] above, wherein the fermented product contains ginsenoside F2 (F2).
[9] The method according to [7] or [8] above, wherein the content ratio (F2 / Re) of ginsenoside F2 (F2) and ginsenoside Re (Re) in the fermented product is 0.001 or more.
[10] Lactobacillus paracasei DNBL 1887 strain (Accession number NITE P-02662).
[11] Lactobacillus paracasei DNBL1888 strain (Accession number NITE P-02663).
本発明では、新規な発酵物を提供できる。このような本発明の発酵物は、通常、「ジンセノシドＦ２」を豊富に含むという優れた効果を奏するものである。また、「ジンセノシドＦ２」を豊富に含むことから、「Ｍ１」の製造原料として好適に用いることができる。 In the present invention, a novel fermented product can be provided. Such a fermented product of the present invention usually has an excellent effect of containing abundant “ginsenoside F2”. Moreover, since it contains abundant “ginsenoside F2”, it can be suitably used as a raw material for producing “M1”.
本発明の発酵物は、ウコギ科薬用ニンジンの、ラクトバチルス・パラカゼイ ＤＮＢＬ１８８７株（受託番号ＮＩＴＥ Ｐ−０２６６２）及び／又はラクトバチルス・パラカゼイ ＤＮＢＬ１８８８株（受託番号ＮＩＴＥ Ｐ−０２６６３）による発酵物であることを特徴とする。なお、ラクトバチルス・パラカゼイ ＤＮＢＬ１８８７株（受託番号ＮＩＴＥ Ｐ−０２６６２）及びラクトバチルス・パラカゼイ ＤＮＢＬ１８８８株（受託番号ＮＩＴＥ Ｐ−０２６６３）は、独立行政法人製品評価技術基盤機構 特許微生物寄託センター（ＮＰＭＤ）（住所：郵便番号２９２−０８１８ 千葉県木更津市かずさ鎌足２−５−８ １２２号室）に、平成３０年（２０１８年）３月６日（受託日）に、受託番号ＮＩＴＥ Ｐ−０２６６２、受託番号ＮＩＴＥ Ｐ−０２６６３としてそれぞれ寄託されている。 The fermented product of the present invention is a fermented product of a carrot family medicinal carrot by Lactobacillus paracasei DNBL 1887 strain (Accession No. NITE P-02662) and / or Lactobacillus paracasei DNBL 1888 strain (Accession No. NITE P-02663) It is characterized by. In addition, Lactobacillus paracasei DNBL 1887 strain (Accession number NITE P-02662) and Lactobacillus paracasei DNBL1888 strain (Accession number NITE P-02663) are independent administrative agency product evaluation technology base patent microorganism deposit center (NPMD) (address) : Postcode 292-0818 Kisarazu City, Kazusa, Kazusa, 2-5-8 122), on March 6, 2018 (contract date), accession number NITE P-02662, accession number NITE Deposited as P-02663, respectively.
ウコギ科薬用ニンジンは、発酵により「ジンセノシドＦ２」を生成可能なニンジンであってもよい。
ウコギ科薬用ニンジンとしては、例えば、オタネニンジン（高麗人参；Ｋｏｒｅａｎ ｇｉｎｓｅｎｇ：Ｐａｎａｘ Ｃ．Ａ．Ｍｅｙｅｒ）、三七ニンジン（Ｐａｎａｘ ｎｏｔｏｇｉｎｓｅｎｇ Ｂｕｒｋ．）、アメリカニンジン（Ｐａｎａｘ ｑｕｉｎｑｕｅｆｏｌｉｕｍ Ｌ．）、竹節ニンジン（Ｐａｎａｘ ｊａｐｏｎｉｃｕｓ Ｃ．Ａ．Ｍｅｙｅｒ）、ヒマラヤニンジン（Ｐａｎａｘ Ｐｓｅｕｄｏ−ｇｉｎｓｅｎｇ Ｑａｌｌ．Ｓｕｂｓｐ．Ｈｉｍａｌａｉｃｕｓ Ｈａｒａ）及びベトナムニンジン（Ｐａｎａｘ Ｖｕｅｔｎａｍｅｎｓｉｓ Ｈａ ｅｔ Ｇｒｕｓｈｖ．）からなる群より選択される少なくとも１種などが挙げられる。本発明の効果が損なわれない範囲内であれば、その他の薬用ニンジンが含まれていてもよい。
The carrot medicinal carrot may be a carrot capable of producing “ginsenoside F2” by fermentation.
For example, ginseng (Korean ginseng: Panax CA Meyer), carrot (Panax quintoseng Burk), American carrot (Panax quinquefolium p. A. Meyer), Himalayan carrot (Panax Pseudo-ginseng Qall. Subsp. Himalaicus Hara) and Vietnamese carrot (Panax Vuenamemensis Ha et Grushv.). Other medicinal carrots may be included as long as the effects of the present invention are not impaired.
上記薬用ニンジンとしては、天然品もその加工品も使用することができる。加工品としては、例えば、薬用ニンジンの乾燥物、裁断物、粉砕物、抽出物、ペースト等が挙げられる。前記乾燥方法、裁断方法、粉砕方法、抽出方法、ペースト状化方法は、従来公知の方法を使用することができる。その大きさは、特に限定されないが、例えば、平均長径が０．２ｍｍ以下に裁断、粉砕、抽出、ペースト状化されることが好ましい。薬用ニンジンとしては、前記した処理を行って調製したものであっても、市販品であってもよい。 As the medicinal carrot, both natural products and processed products thereof can be used. Examples of processed products include dried medicinal carrots, cut products, pulverized products, extracts, and pastes. Conventionally known methods can be used for the drying method, the cutting method, the pulverizing method, the extracting method, and the pasting method. Although the size is not particularly limited, for example, it is preferable that the average major axis is cut, pulverized, extracted, or pasted into a paste of 0.2 mm or less. The medicinal carrot may be prepared by performing the above-described treatment or may be a commercially available product.
本発明において使用される薬用ニンジンの部位は、特に限定されず、どの部位でも使用することができる。例えば、根、茎、葉、花蕾、果実、全草等が挙げられる。これらは１種又は２種以上を使用することができる。好ましくは根等であり、より好ましくは、側根、主根等である。 The site | part of the medicinal carrot used in this invention is not specifically limited, Any site | part can be used. For example, roots, stems, leaves, flower buds, fruits, whole plants and the like can be mentioned. These can use 1 type (s) or 2 or more types. Preferred are roots and the like, and more preferred are side roots and main roots.
薬用ニンジンの発酵物は、前記ニンジンを、微生物を用いて発酵させることにより得られる生産物である。発酵方法は、従来十分に確立されており（例えば、特許３６７８３６２号公報に記載の方法）、本発明もそれに従ってよい。前記発酵は、例えば、薬用ニンジンを含有する培地を常法により減菌処理した後、該培地に微生物を接種し、発酵する方法が好ましく挙げられる。 The fermented medicinal carrot is a product obtained by fermenting the carrot using a microorganism. The fermentation method has been well established in the past (for example, the method described in Japanese Patent No. 3678362), and the present invention may be followed accordingly. The fermentation is preferably performed, for example, by sterilizing a medium containing medicinal carrots by a conventional method, inoculating the medium with a microorganism, and performing fermentation.
本発明において使用される培地は、特に限定されないが、例えば、微生物の培養に通常使用される炭素源、窒素源、ミネラル源等を含むもの等を使用することができ、天然培地又は合成培地等を用いることができる。好ましくは、液体培地を用いる。 The medium used in the present invention is not particularly limited. For example, a medium containing a carbon source, a nitrogen source, a mineral source, etc. that are usually used for culturing microorganisms can be used. Can be used. Preferably, a liquid medium is used.
培地に添加する窒素源としては、特に限定されないが、無機態窒素源としては、例えば、アンモニア、アンモニウム塩等が挙げられ、有機態窒素源としては、例えば、ペプトン、ポリペプトン、尿素、アミノ酸、タンパク質、大豆ペプチド等のペプチド類等が挙げられる。窒素源は、好ましくは、ペプトン、ポリペプトン、ペプチド等である。また、ミネラル源としては、特に限定されないが、酵母エキスや肉エキスの他、Ｋ、Ｐ、Ｍｇ、Ｓ等を含む、例えば、リン酸一水素カリウム、硫酸マグネシウム等が挙げられる。これらの窒素源、ミネラル源は、１種単独で又は２種以上を組み合わせて使用することができる。培地中の窒素源の濃度は、微生物が生育できる通常の濃度であればよく、特に限定されない。培養開始時の窒素源の濃度は、通常は、約０．０５〜１０重量％が好ましく、約０．１〜５重量％がより好ましい。 The nitrogen source to be added to the medium is not particularly limited. Examples of the inorganic nitrogen source include ammonia and ammonium salts. Examples of the organic nitrogen source include peptone, polypeptone, urea, amino acids, and proteins. And peptides such as soybean peptide. The nitrogen source is preferably peptone, polypeptone, peptide or the like. Moreover, it does not specifically limit as a mineral source, In addition to a yeast extract and a meat extract, K, P, Mg, S etc. are contained, for example, potassium monohydrogen phosphate, magnesium sulfate, etc. are mentioned. These nitrogen sources and mineral sources can be used singly or in combination of two or more. The concentration of the nitrogen source in the medium is not particularly limited as long as it is a normal concentration at which microorganisms can grow. The concentration of the nitrogen source at the start of culture is usually preferably about 0.05 to 10% by weight, more preferably about 0.1 to 5% by weight.
前記培地は、前記の窒素源、ミネラル源に加えて、さらに、炭素源、無機質、ｐＨ緩衝剤等を添加しても良い。無機質としては、特に限定されないが、例えば、硫酸アンモニウム、リン酸カリウム、塩化マグネシウム、食塩、鉄、マンガン、モリブデン、各種ビタミン類等が挙げられ、これらは１種単独で又は２種以上を組合せて使用することができる。ｐＨ緩衝剤としては、特に限定されないが、例えば、炭酸カルシウム等が好ましく挙げられる。 In addition to the nitrogen source and mineral source, the medium may further contain a carbon source, an inorganic material, a pH buffering agent, and the like. Examples of inorganic substances include, but are not limited to, ammonium sulfate, potassium phosphate, magnesium chloride, salt, iron, manganese, molybdenum, various vitamins, and the like. These may be used alone or in combination of two or more. can do. Although it does not specifically limit as a pH buffer, For example, a calcium carbonate etc. are mentioned preferably.
薬用ニンジンの使用量は、特に限定されず、その種類、形状、乾燥状態、及び培養条件等に応じて適宜選択され得る。薬用ニンジンと培地全量の重量比（薬用ニンジン／培地全量）としては、好ましくは１／１００〜５０／１００であり、より好ましくは５／１００〜２０／１００であり、更に好ましくは１０／１００〜１５／１００である。本発明において、薬用ニンジンの重量は、内温約１００〜１８０℃で約１〜６時間乾燥させた薬用人参に換算したものであることが好ましい。前記培地は、上述した薬用ニンジン以外の成分や添加剤を含んでも良い。 The amount of medicinal carrot used is not particularly limited, and can be appropriately selected according to the type, shape, dry state, culture conditions, and the like. The weight ratio of medicinal carrot and the total amount of medium (medicinal carrot / total amount of medium) is preferably 1/100 to 50/100, more preferably 5/100 to 20/100, still more preferably 10/100 to 15/100. In the present invention, the weight of the medicinal carrot is preferably converted into medicinal carrots dried at an internal temperature of about 100 to 180 ° C. for about 1 to 6 hours. The medium may contain components and additives other than the medicinal carrot described above.
前記培地のｐＨは、例えば約３〜７とすることが好ましく、約５〜６．５とすることがより好ましい。ｐＨを制御してもよく、酸又はアルカリを用いてｐＨの調整を行うことができる。 The pH of the medium is preferably about 3 to 7, for example, and more preferably about 5 to 6.5. The pH may be controlled, and the pH can be adjusted using an acid or an alkali.
前記培地に前記薬用ニンジンを添加することにより薬用ニンジン含有培地が得られる。該培地を常法により滅菌した後、該培地に微生物を接種して発酵することにより、薬用ニンジンの発酵物が得られる。滅菌方法としては、例えば、加熱滅菌、高圧蒸気滅菌、ろ過滅菌等が挙げられ、これらに限定されることなく従来公知の方法を使用することができる。 A medicinal carrot-containing medium is obtained by adding the medicinal carrot to the medium. After the medium is sterilized by a conventional method, a fermented medicinal carrot is obtained by inoculating the medium with a microorganism and fermenting the medium. Examples of the sterilization method include heat sterilization, high-pressure steam sterilization, filter sterilization, and the like, and a conventionally known method can be used without being limited thereto.
本発明で用いられる微生物は、ラクトバチルス・パラカゼイ ＤＮＢＬ１８８７株（受託番号ＮＩＴＥ Ｐ−０２６６２）及び／又はラクトバチルス・パラカゼイ ＤＮＢＬ１８８８株（受託番号ＮＩＴＥ Ｐ−０２６６３）である。なお、本発明においては、本発明の効果を損なわない範囲内であれば、その他の公知の乳酸菌を用いてもよい。 The microorganism used in the present invention is Lactobacillus paracasei DNBL1887 strain (Accession No. NITE P-02662) and / or Lactobacillus paracasei DNBL1888 strain (Accession No. NITE P-02663). In the present invention, other known lactic acid bacteria may be used as long as the effects of the present invention are not impaired.
ラクトバチルス属菌の使用量は、前記の特定菌体を用いるのであれば特に限定はなく、当業者によって適宜設定され得る。 The amount of Lactobacillus genus used is not particularly limited as long as the specific cells are used, and can be appropriately set by those skilled in the art.
かかるラクトバチルス属菌を用いて薬用ニンジンを発酵させる。 The medicinal carrot is fermented using the Lactobacillus sp.
具体的には、例えば、前記した薬用ニンジン含有培地に、ラクトバチルス・パラカゼイ ＤＮＢＬ１８８７株（受託番号ＮＩＴＥ Ｐ−０２６６２）及び／又はラクトバチルス・パラカゼイ ＤＮＢＬ１８８８株（受託番号ＮＩＴＥ Ｐ−０２６６３）を添加し、好ましくは２５〜３７℃、より好ましくは２８〜３３℃で発酵させる。発酵時間は、仕込み液の組成、植菌量、発酵温度などに応じて適切に設定することが可能であり、例えば、２〜２１日、７〜１４日などが例示される。なお、前記菌体を双方使用する場合は、別々に培地に添加して発酵させても、同一の培地に組み合わせて添加して発酵させてもよい。 Specifically, for example, Lactobacillus paracasei DNBL1887 strain (Accession number NITE P-02662) and / or Lactobacillus paracasei DNBL1888 strain (Accession number NITE P-02663) is added to the above-described medicinal carrot-containing medium, Preferably it is made to ferment at 25-37 degreeC, More preferably, it is 28-33 degreeC. The fermentation time can be appropriately set according to the composition of the preparation liquid, the amount of inoculation, the fermentation temperature, and the like, and examples thereof include 2 to 21 days and 7 to 14 days. In addition, when using both the said microbial cells, it may be separately added to a culture medium and fermented, or may be added and fermented in combination with the same culture medium.
以上のようにして得られた発酵物には、ジンセノシドＦ２（Ｆ２）が多く含まれている。このような発酵物は、ジンセノシドＦ２（Ｆ２）含有発酵物と称することができる。
そのため、本発明には、ジンセノシドＦ２を含有する発酵物（発酵組成物）も包含する。
このような発酵物は、その製造方法において特に限定されないが、上記の特定の微生物を用いることにより、効率良く製造しうる。
The fermented product obtained as described above contains a large amount of ginsenoside F2 (F2). Such a fermented product can be referred to as a ginsenoside F2 (F2) -containing fermented product.
Therefore, the present invention also includes a fermented product (fermented composition) containing ginsenoside F2.
Such a fermented product is not particularly limited in its production method, but can be efficiently produced by using the above-mentioned specific microorganism.
発酵物中、ジンセノシドＦ２（Ｆ２）とジンセノシドＲｅ（Ｒｅ）の含有量比（Ｆ２／Ｒｅ）は、好ましくは０．００１以上、より好ましくは０．０１以上、更に好ましくは０．１以上、更に好ましくは１以上である。 In the fermented product, the content ratio (F2 / Re) of ginsenoside F2 (F2) and ginsenoside Re (Re) is preferably 0.001 or more, more preferably 0.01 or more, still more preferably 0.1 or more, Preferably it is 1 or more.
得られた発酵物に対しては、所望により、ろ過、遠心分離、濃縮、限外ろ過、凍結乾燥、粉末化、分画等の処理を公知の方法に従って行ってもよく、目的物（例えば、Ｆ２を含む画分）そのものを単離して又は培地を含むものをそのまま製剤化したり、飲食品（健康食品）等の原材料に用いることが可能な形態に調製したりしてもよい。よって、本発明の発酵物の一態様として前記処理物が含まれる。 If desired, the obtained fermented product may be subjected to treatments such as filtration, centrifugation, concentration, ultrafiltration, lyophilization, pulverization, fractionation and the like according to a known method, for example, a target product (for example, The F2 containing fraction) itself may be isolated or the medium containing the medium may be formulated as it is, or may be prepared in a form that can be used for raw materials such as foods and drinks (health foods). Therefore, the said processed material is contained as one aspect | mode of the fermented material of this invention.
本発明はまた、前記発酵物を含有してなる組成物を提供する。かかる発酵物又は組成物は、例えば、抗腫瘍、抗肥満、育毛、抗健忘症、肝保護作用、抗動脈硬化、老化防止、抗高脂血症、抗血栓、血圧降下作用、免疫機能改善、血糖値低下作用、鎮痛作用、強心作用、抗炎症作用、末梢血流改善作用、止血作用、抗酸化作用、抗筋委縮作用、骨量増加作用及び抗ストレス用から選ばれる少なくとも１種の作用の発揮を期待して好適に使用することができる。 The present invention also provides a composition comprising the fermented product. Such fermented products or compositions are, for example, anti-tumor, anti-obesity, hair growth, anti-amnesia, hepatoprotective action, anti-arteriosclerosis, anti-aging, anti-hyperlipidemic, anti-thrombotic, anti-thrombotic action, immune function improvement, At least one action selected from a blood glucose lowering action, an analgesic action, a cardiotonic action, an anti-inflammatory action, a peripheral blood flow improving action, a hemostatic action, an antioxidant action, an antimuscular atrophy action, a bone mass increasing action and an anti-stress effect It can be suitably used with the expectation of exhibiting.
本発明の発酵物又は組成物は、ジンセノシドＦ２（Ｆ２）やジンセノシドＲｅ（Ｒｅ）以外に、ジンセノシドＲｂ１、ジンセノシドＲｂ２、ジンセノシドＲｃ、ジンセノシドＲｄ、ジンセノシドＲｇ１、ジぺノシドＸＶＩＩなどを含有してもよい。それらの含有量は、本発明の効果を損なわない範囲内であれば、特に限定されない。 The fermented product or composition of the present invention may contain ginsenoside Rb1, ginsenoside Rb2, ginsenoside Rc, ginsenoside Rd, ginsenoside Rg1, dipenoside XVII, etc. in addition to ginsenoside F2 (F2) and ginsenoside Re (Re). . Their content is not particularly limited as long as it does not impair the effects of the present invention.
本発明の組成物は、本発明の発酵物を体内に摂取できるのであれば、その形態は限定されない。例えば、錠剤、散剤、顆粒剤、カプセル剤、液剤、乳剤、エリキシル剤、懸濁剤、シロップ剤、トローチ剤、吸入剤、坐剤、注射剤、軟膏剤、眼軟膏剤、点眼剤、点鼻剤、点耳剤、パップ剤、ローション剤等の経口製剤又は非経口製剤として製剤化することができる。 The form of the composition of the present invention is not limited as long as the fermented product of the present invention can be taken into the body. For example, tablets, powders, granules, capsules, solutions, emulsions, elixirs, suspensions, syrups, troches, inhalants, suppositories, injections, ointments, eye ointments, eye drops, nose drops It can be formulated as an oral preparation or a parenteral preparation such as an agent, ear drops, cataplasm, lotion and the like.
本発明の発酵物又は組成物の投与条件は、その形態や投与目的、当該組成物の投与対象の種類、年齢、体重、症状によって適宜設定され一定ではない。例えば、Ｆ２の有効ヒト投与量として、一日当たり、好ましくは０．０１〜１００ｍｇ／ｋｇ程度、より好ましくは１〜５０ｍｇ／ｋｇ程度となるよう設定することができる。投与は、所望の投与量範囲内において、１日内において単回で又は数回に分けて行ってもよい。投与期間も任意である。 The administration conditions of the fermented product or composition of the present invention are appropriately set depending on the form and purpose of administration, the type of administration target of the composition, age, weight, and symptoms, and are not constant. For example, the effective human dose of F2 can be set to be about 0.01 to 100 mg / kg, more preferably about 1 to 50 mg / kg per day. Administration may be performed once or divided into several times within one day within a desired dose range. The administration period is also arbitrary.
本発明の発酵物又は組成物の投与対象としては、好ましくは抗腫瘍、抗肥満、育毛、抗健忘症、肝保護作用、抗動脈硬化、老化防止、抗高脂血症、抗血栓、血圧降下作用、免疫機能改善、血糖値低下作用、鎮痛作用、強心作用、抗炎症作用、末梢血流改善作用、止血作用、抗酸化作用、抗筋委縮作用、骨量増加作用及び抗ストレス用から選ばれる少なくとも１種の作用の発揮を必要とするヒトであるが、ウシ、ウマ、ヤギ等の家畜動物、イヌ、ネコ、ウサギ等のペット動物、又は、マウス、ラット、モルモット、サル等の実験動物であってもよい。 The fermented product or composition of the present invention is preferably administered with antitumor, anti-obesity, hair growth, anti-amnesia, hepatoprotective action, anti-arteriosclerosis, anti-aging, anti-hyperlipidemia, anti-thrombosis, blood pressure lowering Action, immune function improvement, blood sugar level lowering action, analgesic action, cardiotonic action, anti-inflammatory action, peripheral blood flow improving action, hemostasis action, antioxidant action, antimuscular atrophy action, bone mass increasing action and anti-stress use It is a human who needs to exhibit at least one kind of action, but it is a domestic animal such as a cow, horse or goat, a pet animal such as a dog, cat or rabbit, or a laboratory animal such as a mouse, rat, guinea pig or monkey. There may be.
上記形態を有する本発明の組成物は、本発明の発酵物を含有するものであれば、常法に従って、製剤分野等において通常使用される担体、基剤、及び／又は添加剤等を本発明の目的を達成する範囲内で適宜配合して調製することができる。具体的には、例えば、賦形剤、結合剤、滑沢剤、着色剤、矯味矯臭剤や、及び必要により安定化剤、乳化剤、吸収促進剤、界面活性剤、pH調製剤、防腐剤、抗酸化剤等を使用することができ、一般に医薬品製剤の原料として用いられる成分を、本発明の効果を損なわない範囲内で適量配合して常法により製剤化される。本発明の発酵物の含有量は、剤型、投与方法、担体等により異なるが、本発明の組成物中、通常０．０１〜１００重量％、好ましくは０．１〜９５重量％である。 As long as the composition of the present invention having the above-described form contains the fermented product of the present invention, the carrier, base, and / or additive or the like usually used in the pharmaceutical field and the like according to the conventional method are used in the present invention. In the range which achieves the objective, it can mix | blend suitably and can prepare. Specifically, for example, excipients, binders, lubricants, colorants, flavoring agents, and if necessary stabilizers, emulsifiers, absorption promoters, surfactants, pH adjusters, preservatives, Antioxidants and the like can be used, and the components generally used as raw materials for pharmaceutical preparations are blended in appropriate amounts within a range that does not impair the effects of the present invention, and are formulated by conventional methods. The content of the fermented product of the present invention varies depending on the dosage form, administration method, carrier and the like, but is usually 0.01 to 100% by weight, preferably 0.1 to 95% by weight in the composition of the present invention.
また、本発明は、ウコギ科薬用ニンジンを、ラクトバチルス・パラカゼイ ＤＮＢＬ１８８７株（受託番号ＮＩＴＥ Ｐ−０２６６２）及び／又はラクトバチルス・パラカゼイ ＤＮＢＬ１８８８株（受託番号ＮＩＴＥ Ｐ−０２６６３）により発酵させることを特徴とする、発酵物の製造方法を提供する。ここで、原料の薬用ニンジンや使用する微生物、その発酵方法としては、本発明の発酵物を参照して設定することができる。このような方法によれば、前記のように、ジンセノシドＦ２を含有する発酵物を効率よく得やすい。ジンセノシドＦ２の含有割合等もまた、前記と同様であってもよい。 In addition, the present invention is characterized by fermenting a carrot medicinal carrot with Lactobacillus paracasei DNBL 1887 strain (Accession No. NITE P-02662) and / or Lactobacillus paracasei DNBL 1888 strain (Accession No. NITE P-02663). A method for producing a fermented product is provided. Here, the raw material medicinal carrot, the microorganism to be used, and the fermentation method can be set with reference to the fermented product of the present invention. According to such a method, as described above, it is easy to efficiently obtain a fermented product containing ginsenoside F2. The content ratio of ginsenoside F2 and the like may also be the same as described above.
以下、実施例により本発明を詳細に説明するが、本発明はこれらに限定されるものではない。 EXAMPLES Hereinafter, although an Example demonstrates this invention in detail, this invention is not limited to these.
試験例１
（１） １ｇのオタネニンジン末に０．２ｇの炭酸カルシウムを加え、先ずミリＱ１０ｍＬに懸濁して１２１℃１５分加熱滅菌を実施した。
（２） その後滅菌ミリＱ１０ｍＬを更に加え、計２０ｍＬとした。
（３） そこへ乳酸菌を予め一般乳酸菌培地（日水）５ｍＬで３日間培養した菌体を５００μＬ加えて、良く混ぜ、３７℃にて静置して発酵を開始した。
（４） 開始後９日目と１９日目の時点でピペットでよく懸濁したもの５００μＬを取り、等量のエタノールを加えてボルテクスを５秒行い１５０００ｒｐｍで５分遠心を実施した。
（５） その上清より７μＬとり、ＴＬＣプレートにスポットして、展開を実施した。展開液は酢酸エチル：メタノール：水＝１４：５：４（ｖ／ｖ／ｖ）を使用した。その後検出液を噴霧して各成分の検出を実施した。バニリン３ｇをエタノール３０ｍＬに溶解したものに３ｍｏｌ／Ｌの硫酸を１００ｍＬ添加したものを検出液として使用した。なお、ジンセノシド標品の発酵処理は市販されているジンセノシドＲｂ１標品を発酵基質として各種菌体ペレットに加えて同様の操作を行った。
（６） また、発酵開始後１９日目の時点でＴＬＣへ供した上清をＨＰＬＣへ供し定量分析を実施した。分析条件としては、カラム（ＹＭＣ−Ｐａｃｋ ＯＤＳ−Ａ ２５０×４．６ｍｍ）、カラムオーブン４０℃、Ａ液：水、Ｂ液：アセトニトリル、流速：１ｍＬ／分、検出：２０３ｎｍ、で実施し、ジンセノシドＦ２の分析ではＢ液５０％、ジンセノシドＲｅの分析ではＢ液２０％のイソクラチック条件下で実施した。
Test example 1
(1) 0.2 g of calcium carbonate was added to 1 g of ginseng powder, and the suspension was first suspended in 10 mL of milli Q and sterilized by heating at 121 ° C. for 15 minutes.
(2) Thereafter, 10 mL of sterile MilliQ was further added to make a total of 20 mL.
(3) 500 μL of lactic acid bacteria previously cultured in 5 mL of general lactic acid bacteria medium (day water) for 3 days was added thereto, mixed well, and allowed to stand at 37 ° C. to initiate fermentation.
(4) On the 9th and 19th day after the start, 500 μL of the well suspended solution was taken, and an equal volume of ethanol was added, vortexed for 5 seconds, and centrifuged at 15000 rpm for 5 minutes.
(5) 7 μL was taken from the supernatant and spotted on a TLC plate for development. The developing solution used was ethyl acetate: methanol: water = 14: 5: 4 (v / v / v). Thereafter, the detection liquid was sprayed to detect each component. A solution obtained by adding 100 mL of 3 mol / L sulfuric acid to 3 g of vanillin dissolved in 30 mL of ethanol was used as a detection solution. In addition, the fermentation process of the ginsenoside sample performed the same operation, adding the commercially available ginsenoside Rb1 sample as a fermentation substrate to various cell pellets.
(6) Moreover, the supernatant used for TLC at the time of the 19th day after fermentation start was used for HPLC, and the quantitative analysis was implemented. The analysis conditions were as follows: column (YMC-Pack ODS-A 250 × 4.6 mm), column oven 40 ° C., liquid A: water, liquid B: acetonitrile, flow rate: 1 mL / min, detection: 203 nm, ginsenoside The analysis of F2 was performed under isocratic conditions of solution B 50%, and ginsenoside Re was analyzed with solution B 20%.
図１に示すように、ラクトバチルス・パラカゼイ ＤＮＢＬ１８８７株（受託番号ＮＩＴＥ Ｐ−０２６６２）及びラクトバチルス・パラカゼイ ＤＮＢＬ１８８８株（受託番号ＮＩＴＥ Ｐ−０２６６３）を用いて発酵することで、Ｆ２の生成量が飛躍的に増加した。 As shown in FIG. 1, the fermentation amount of Lactobacillus paracasei DNBL 1887 (accession number NITE P-02662) and Lactobacillus paracasei DNBL1888 strain (accession number NITE P-02663) is greatly increased. Increased.
表１〜２に示すように、ラクトバチルス・パラカゼイ ＤＮＢＬ１８８７株（受託番号ＮＩＴＥ Ｐ−０２６６２）及びラクトバチルス・パラカゼイ ＤＮＢＬ１８８８株（受託番号ＮＩＴＥ Ｐ−０２６６３）は効率よくジンセノシドＦ２への代謝が進んでおり、それぞれ原末乾燥固形物重量で１．６８％、１．３６％の含有量となった。一方、比較で用いたラクトバチルス・プランタラムは全くジンセノシドＦ２を生産しなかった。 As shown in Tables 1-2, Lactobacillus paracasei DNBL 1887 strain (Accession number NITE P-02662) and Lactobacillus paracasei DNBL1888 strain (Accession number NITE P-02663) are efficiently metabolized to ginsenoside F2. And 1.68% and 1.36%, respectively, in terms of the weight of the solid powder dry solids. On the other hand, Lactobacillus plantarum used for comparison did not produce ginsenoside F2.
本発明では新規な発酵物を提供できる。このような発酵物は、通常、ジンセノシドＦ２（Ｆ２）が多く含まれており、例えば、抗腫瘍、抗肥満、育毛、抗健忘症、肝保護作用、抗動脈硬化、老化防止、抗高脂血症、抗血栓、血圧降下作用、免疫機能改善、血糖値低下作用、鎮痛作用、強心作用、抗炎症作用、末梢血流改善作用、止血作用、抗酸化作用、抗筋委縮作用、骨量増加作用及び抗ストレス用から選ばれる少なくとも１種の作用の発揮を期待する疾患に好適に用いることができる。 In the present invention, a novel fermented product can be provided. Such fermented products usually contain a large amount of ginsenoside F2 (F2). For example, anti-tumor, anti-obesity, hair growth, anti-amnesia, hepatoprotective action, anti-arteriosclerosis, anti-aging, anti-hyperlipidemia Disease, antithrombosis, blood pressure lowering effect, immune function improvement, blood sugar level lowering effect, analgesic effect, cardiotonic effect, anti-inflammatory effect, peripheral blood flow improving effect, hemostatic effect, antioxidant effect, antimuscular atrophy, bone mass increasing effect And can be suitably used for diseases that are expected to exhibit at least one action selected from anti-stress use.
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