JP2014205634A - External preparation for skin - Google Patents
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Abstract
Description
本発明は、皮膚外用剤に係り、特に皮膚の保湿に好適な皮膚外用剤に関する。 The present invention relates to an external preparation for skin, and particularly relates to an external preparation for skin suitable for moisturizing skin.
本発明において、穀類とは、大豆、米、麦、とうもろこしやこれらの粕等を意味し、本発明はこれらの穀類の少なくとも１種を原料基質として使用する。 In the present invention, cereal means soybeans, rice, wheat, corn, potatoes and the like, and the present invention uses at least one of these cereals as a raw material substrate.
本出願人は、すでに穀物由来の生成物について、その製造方法を特許第２６９６０５７号公報において提案している。この製造方法によれば、穀類を原料として麹菌によって製麹させて蛋白質を分解し、その後に加水分解することによって得られた生成物においては、穀類の状態を固形状にしたままで穀類中のフィチン酸が除去され、生成物中に含有されているビタミンＢ類等の活性が高く維持され、生成物中に含有されているミネラルの吸収も容易とされ、更にその吸収も促進可能とされている。このような穀類由来の生成物は人の健康によく作用するものである。 The present applicant has already proposed a method for producing a grain-derived product in Japanese Patent No. 2696057. According to this production method, in a product obtained by producing a cereal using koji mold as a raw material, decomposing protein, and then hydrolyzing it, the cereal state remains solid while the cereal is in a solid state. Phytic acid is removed, the activity of vitamin B and the like contained in the product is maintained high, the absorption of minerals contained in the product is facilitated, and the absorption can also be promoted. Yes. Such cereal-derived products work well for human health.
また、本出願人は、すでに穀類の１種である大豆由来のイソフラボンアグリコンについて、その製造方法を特許第３０１４１４５号公報において、豆類を原料として麹菌によって発酵させて蛋白質を分解し、その後に加水分解することによって得られた生成物には大豆イソフラボングリコシドが麹菌の酵素（β−グリコシダーゼ）でイソフラボンアグリコンに変化されるため、イソフラボンアグリコンがリッチに含有されており、この生成物が人間や哺乳類に対して薬効等の生体活性を促進する作用をなすことを提案している。 In addition, the applicant of the present invention disclosed a method for producing isoflavone aglycone derived from soybean, which is a kind of cereal, in Japanese Patent No. 3014145, using bean as a raw material to ferment by a koji mold, followed by hydrolysis, followed by hydrolysis. The soy isoflavone glycoside is converted into an isoflavone aglycone by the koji mold enzyme (β-glycosidase), so that the isoflavone aglycone is rich in this product. It has been proposed to promote biological activities such as medicinal effects.
近年、健康的で豊かな生活を送るために肌の健康を維持することが注目されている。そこで、シワの防止、肌のハリやツヤの改善に効果を発揮し、保湿性を向上させることを目的とした種々の皮膚外用剤が提案されている（例えば、特許文献３参照）。 In recent years, attention has been focused on maintaining the health of the skin in order to lead a healthy and prosperous life. Therefore, various skin external preparations have been proposed that are effective in preventing wrinkles, improving skin firmness and gloss, and improving moisture retention (see, for example, Patent Document 3).
しかしながら、種々の改善策が提案されているが、肌の保湿性に優れており、副作用が無く、安全で、安価な皮膚外用剤は未だに得られていない。 However, various improvement measures have been proposed, but skin external preparations having excellent skin moisture retention, no side effects, and safe and inexpensive have not been obtained yet.
本発明はこれらの点に鑑みてなされたものであり、肌の保湿性に優れており、副作用が無く、安全で、安価な皮膚外用剤を提供することを目的とする。 The present invention has been made in view of these points, and an object of the present invention is to provide a skin external preparation that is excellent in skin moisture retention, has no side effects, and is safe and inexpensive.
本発明者は鋭意研究し、本出願人が特許第２６９６０５７号および特許第３０１４１４５号において既に提案している生成物（「イムバランス」（登録商標））について、その成分と作用とを究明し、当該生成物並びに更に乳酸菌を含有している生成物内に糖セラミド類が含有されていることを発見して本発明を完成させた。 The present inventor has intensively studied, investigated the components and actions of the product ("Imbalance" (registered trademark)) that the applicant has already proposed in Japanese Patent Nos. 2696057 and 3014145, The present invention was completed by discovering that sugar ceramides were contained in the product as well as a product containing lactic acid bacteria.
このようにしてなされた本発明の皮膚外用剤は、穀類に麹菌を接種して製麹し、この製麹処理による生成物に加水することにより当該生成物中の組成要素を加水分解した固体組成物であって、糖セラミド類を含有する固体組成物を含有することを特徴とする。 The external preparation for skin of the present invention thus made is a solid composition obtained by inoculating cereals with koji molds and kneading them, and then hydrolyzing the components of the products by adding water to the products obtained by this koji making process. And a solid composition containing sugar ceramides.
また、前記固体生成物は、前記生成物に対する麹菌による代謝物を含むことを特徴とする。 Further, the solid product includes a metabolite produced by Aspergillus oryzae with respect to the product.
また、本発明の皮膚外用剤は、穀類に麹菌を接種して製麹し、この製麹処理による生成物に加水することにより当該生成物中の組成要素を加水分解した固体組成物であって、糖セラミド類および乳酸菌を含有する固体組成物を含有することを特徴とする。 Further, the external preparation for skin of the present invention is a solid composition obtained by inoculating cereals with koji mold and making a koji, and then hydrolyzing the components in the product by adding water to the product by this koji making process. And a solid composition containing sugar ceramides and lactic acid bacteria.
この糖セラミド類および乳酸菌を含有する固体組成物は、前記生成物に対する麹菌および乳酸菌による代謝物を含むものであり、当該代謝物としては、黄色ブドウ球菌および大腸菌の増殖を抑制するバクテリオシンや、Ｔｈ２サイトカインの生産を抑制する機能を備えた麹多糖類を含むことを特徴とする。 The solid composition containing the saccharide ceramides and lactic acid bacteria contains a metabolite by gonococci and lactic acid bacteria with respect to the product, and as the metabolite, bacteriocin that suppresses the growth of S. aureus and E. coli, It comprises a polysaccharide having a function of suppressing the production of Th2 cytokines.
また、皮膚外用剤が化粧品であることを特徴とする。 The skin external preparation is a cosmetic.
このようにして形成されている本発明の皮膚外用剤は、糖セラミド類による保湿作用が発揮されて、肌の保湿性に優れており、副作用が無く、安全で、安価な皮膚外用剤を得ることができる。 The external preparation for skin of the present invention thus formed exhibits a moisturizing action by sugar ceramides, has excellent skin moisturizing properties, has no side effects, and provides a safe and inexpensive external preparation for skin. be able to.
本発明の皮膚外用剤によれば、肌の保湿性に優れており、副作用が無く、安全で、安価な皮膚外用剤を提供することができるという優れた効果を奏する。 According to the external preparation for skin of the present invention, the skin moisturizing property is excellent, and there is no side effect, and there is an excellent effect that a safe and inexpensive external preparation for skin can be provided.
以下、図１および図２を用いて本発明の皮膚外用剤の実施形態について説明する。 Hereinafter, embodiments of the external preparation for skin of the present invention will be described with reference to FIGS. 1 and 2.
本実施形態においては、本出願人が特許第２６９６０５７号および特許第３０１４１４５号において既に提案している生成物（イムバランス）であって、「原料となる穀類の１種である大豆等の豆類に対して麹菌を用いて製麹処理を施すとともにその生成物に対して加水分解処理を施して、蛋白質が分解されており、イソフラボンアグリコンを多量に含有する生成物やフイチン酸が完全に分解除去された固体組成物または当該固体生成物に乳酸菌を含有するものであって糖セラミド類を含有している固体組成物含有する皮膚外用剤」を発明品とする。 In the present embodiment, the product (imbalance) already proposed by the present applicant in Patent No. 2696057 and Patent No. 3014145, which is “to bean such as soybean that is one kind of cereal as a raw material” On the other hand, the koji mold is used to make a koji and the product is hydrolyzed to decompose the protein, and the product and phytic acid containing a large amount of isoflavone aglycone are completely decomposed and removed. In addition, the “external preparation for skin containing a solid composition containing lactic acid bacteria and containing sugar ceramides” in the solid composition or the solid product is an invention.
図１は前記特許第２６９６０５７号および特許第３０１４１４５号において提案している生成物であって、豆類の１種である脱脂大豆のイソフラボン化合物の配糖体を分解して、アグリコンを多量に含むイソフラボン化合物を生成した生成物の製造方法の１実施形態および同時に脱脂大豆中のフィチン酸を除去した生成物の製造方法の１実施形態を示す工程図である。 FIG. 1 is a product proposed in the aforementioned Patent Nos. 2696057 and 3014145, which is an isoflavone containing aglycone in a large amount by decomposing a glycoside of an isoflavone compound of defatted soybean which is a kind of beans. It is process drawing which shows one Embodiment of the manufacturing method of the product which produced | generated the compound, and 1 embodiment of the manufacturing method of the product which removed the phytic acid in defatted soybean simultaneously.
図１の工程に沿って説明すると、先ず脱脂大豆を蒸煮する。この蒸煮を施すことにより、麹菌の増殖が容易となる。また、この脱脂大豆の蒸煮は製造目的等に応じてバッチ式や連続式で行うと良い。 If it demonstrates along the process of FIG. 1, first, defatted soybean will be cooked. By performing this cooking, the growth of koji molds is facilitated. The defatted soybeans may be cooked batchwise or continuously depending on the purpose of manufacture.
そして、この蒸煮が終了した脱脂大豆を一旦冷却して、脱脂大豆中の水分量を麹菌が増殖可能な量（例えば、３７重量％）とさせる。 Then, the defatted soybean that has been cooked is once cooled, and the amount of water in the defatted soybean is adjusted to an amount that allows the koji mold to grow (for example, 37% by weight).
このようにして水分量を整えられた脱脂大豆に対して、本発明方法が以下のようにして行なわれる。 Thus, the method of this invention is performed as follows with respect to the defatted soybean in which the moisture content was adjusted.
即ち、蒸煮が終了した脱脂大豆を殺菌し、冷却した後、脱脂大豆と麹菌との配合割合を、例えば脱脂大豆を４００ｋｇに対して麹菌胞子を８×１０7 個／ｇに調整した種麹（精白米にて調整）を２００ｇを混合した。更に、製麹のスタート時には３２℃に冷却した後、品温が４０℃になるまで通風しないで４０℃になった時点で通風しながら、温度上昇を抑えた。スタートから約１７時間後の最初の撹拌（盛工程）を行った。脱脂大豆の熱を冷まし、撹拌後品温が３５℃前後になるように通風しながら、温度をコントロールをした。次いで約８時間後に２回目の撹拌（仲工程）を行い、熱を冷ました。再び品温を通風で３５℃前後にコントロールし、更に約１６時間後に３回目の撹拌（仕舞工程）を前回同様に行った。その後は品温が約３８℃になるように通風しながら、温度コントロールし、スタートから４８時間後に製麹を終了させた。製麹終了後、水分含量が５０％になるように撹拌しながら、水分調整を行い、品温が約５０℃になるように加温後、４８時間以上麹菌の酵素で脱脂大豆中のイソフラボン化合物の大部分がアグリコン体になるまで加水分解した。表１に示す通り本発明による処理により脱脂大豆のイソフラボン化合物はアグリコン体のダイゼインが主体となるように多量に得られた。 That is, after defatted soybeans that have been cooked are sterilized and cooled, the blending ratio of defatted soybeans and koji molds, for example, 400 kg of defatted soybeans, and koji molds adjusted to 8 × 10 7 spores / g 200 g of the mixture was adjusted with polished rice. In addition, after the start of the iron making, after cooling to 32 ° C., the temperature rise was suppressed while ventilating when the temperature reached 40 ° C. until the product temperature reached 40 ° C. About 17 hours after the start, the first stirring (pick-up process) was performed. The heat of the defatted soybean was cooled, and the temperature was controlled while ventilating so that the product temperature was about 35 ° C. after stirring. Next, about 8 hours later, the second stirring (middle process) was performed to cool the heat. Again, the product temperature was controlled to around 35 ° C. with ventilation, and after about 16 hours, the third stirring (closing process) was performed in the same manner as the previous time. Thereafter, the temperature was controlled while ventilating so that the product temperature was about 38 ° C., and the ironmaking was finished 48 hours after the start. After completion of the koji making, the water content is adjusted while stirring so that the water content becomes 50%, and the product is heated so that the product temperature becomes about 50 ° C., and then the isoflavone compound in the defatted soybean with the koji mold enzyme for 48 hours or more. It hydrolyzed until most of it became an aglycon body. As shown in Table 1, a large amount of defatted soybean isoflavone compound was obtained by treatment according to the present invention so that aglycone daidzein was the main component.
この製麹に用いる麹菌としては、古くからの日本独特の発酵食品やテンペに用いられている麹菌であり、食品として安全なアスペルギルス・ウサミ、アスペルギルス・カワチ、アスペルギルス・アワモリ、アスペルギルス・サイトイ、アスペルギルス・オリゼー、アスペルギルス・ニガー等アスペルギルス属およびリゾープス属等からなる麹菌を用いるとよい。 As the koji mold used in this koji making, it is koji mold that has been used in traditional Japanese fermented foods and tempeh. Aspergillus oryzae or Aspergillus niger or other Aspergillus genus, Rhizopus genus or the like may be used.
この発酵時間については、使用する麹菌の種類に応じて、少なくとも２４時間以上であり、脱脂大豆中のイソフラボン化合物の配糖体を十分に分解させるに十分な発酵時間とするとよい。 The fermentation time is at least 24 hours or longer depending on the type of koji mold used, and is preferably set to a fermentation time sufficient to sufficiently decompose the glycoside of the isoflavone compound in the defatted soybean.
この蛋白質の加水分解については、使用する麹菌の種類に応じて、脱脂大豆中のイソフラボン化合物の配糖体を十分に低減させるに十分な加水分解時間ならびに加水分解温度とするとよい。 About hydrolysis of this protein, it is good to set it as hydrolysis time and hydrolysis temperature sufficient to fully reduce the glycoside of the isoflavone compound in defatted soybean according to the kind of koji mold used.
このようにすれば、発酵の初期において有機酸を生成して脱脂大豆中の雑菌の増殖を抑制し、２次汚染の心配がなくなり、脱脂大豆を原料とした固形生成物を大量生産することができる。また、低水分としなくともイソフラボン化合物の配糖体を十分に低減させる処理を施すことができる。 In this way, it is possible to produce organic acids at the initial stage of fermentation to suppress the growth of germs in the defatted soybean, eliminate the risk of secondary contamination, and mass-produce solid products made from defatted soybean. it can. Moreover, even if it does not set it as low moisture, the process which fully reduces the glycoside of an isoflavone compound can be given.
加水分解の終了後には、組成物を乾燥させ、必要に応じて粉砕若しくは滅菌することにより固形生成物を得る。 After completion of hydrolysis, the composition is dried and, if necessary, ground or sterilized to obtain a solid product.
この固形生成物には、前記の一連の生産工程を経ることにより糖セラミド類が０．５５ｍｇ／ｇの割合で含有されていることがわかった。 This solid product was found to contain sugar ceramides at a rate of 0.55 mg / g through the series of production steps described above.
更に、この固形生成物には前記の一連の生産工程を経ることにより乳酸菌が増殖されている固体生成物があることがわかった。 Furthermore, it was found that this solid product includes a solid product in which lactic acid bacteria are grown through the above-described series of production steps.
その乳酸菌の種類を各種の検出培地を用いて増殖させて検出したところ、乳酸菌はペディオコッカス属（Pediococcus sp.）とエントロコッカス属（Entrococcus sp.）とを含み、当該ペディオコッカス属およびエントロコッカス属の菌数の比が約３：１であることが判明した。 When the type of the lactic acid bacteria was grown and detected using various detection media, the lactic acid bacteria included Pediococcus sp. And Entrococcus sp., And the Pediococcus sp. It was found that the ratio of the number of Coccus bacteria was about 3: 1.
また、更に本実施形態の固形生成物には、麹菌および乳酸菌が増殖することによる固形組成物の代謝成分として、例えば、麹多糖類（アラビノース（含有量：４１．４％）、キシロース（含有量：１０．４％）、マンノース（含有量：４．０％）およびその他の成分）、ペプチド等が産生されていることがわかった。 Further, in the solid product of the present embodiment, as a metabolic component of the solid composition resulting from the growth of gonococci and lactic acid bacteria, for example, potato polysaccharides (arabinose (content: 41.4%), xylose (content) 10.4%), mannose (content: 4.0%) and other components), peptides and the like were produced.
これらの麹多糖類は、後述するように、Ｔｈ２サイトカインの生産を抑制する機能を備えており、また、ペプチドには黄色ブドウ球菌および大腸菌の増殖を抑制するバクテリオシンが含まれていることがわかった。 As will be described later, these sputum polysaccharides have a function of suppressing the production of Th2 cytokines, and the peptides contain bacteriocin that suppresses the growth of S. aureus and E. coli. It was.
本発明の皮膚外用剤は、前記のようにして生成された固形生成物を皮膚外用剤の基剤に対して所定重量（例えば、０．３〜１．０％）含有させることにより生成される。 The external preparation for skin of the present invention is produced by containing the solid product produced as described above in a predetermined weight (for example, 0.3 to 1.0%) with respect to the base of the external preparation for skin. .
この皮膚外用剤の基剤としては、例えばクリームやジェル等を挙げることができ、外用剤を化粧品とする場合には、例えば、ナノスフェアクリーム、キサンタンガム、ジェル等を挙げることができる。 Examples of the base for the external preparation for skin include creams and gels. For example, when the external preparation is used as a cosmetic, nanosphere cream, xanthan gum, gels and the like can be used.
このようにして形成されている本発明の皮膚外用剤（化粧品を含む）を、複数の被検者の顔に塗布してその効能を試験したところ、全被検者において、保湿効果の向上を検出し、肌の健康が促進されたことを確認した。 When the external preparation for skin of the present invention (including cosmetics) thus formed was applied to the faces of a plurality of subjects and tested for its effectiveness, the moisturizing effect was improved in all subjects. Detected and confirmed that skin health was promoted.
更に、皮膚外用剤を、複数の被検者の足の踵のひび割れ部分（長さ１０ｍｍ×幅０．５ｍｍ×深さ１．５ｍｍ）に塗布してその効能を試験したところ、２週間後にひび割れ部分がほぼ半分に治癒され、５週間後には全快されたことを確認した。 Furthermore, when the external preparation for skin was applied to the cracked portions (length 10 mm × width 0.5 mm × depth 1.5 mm) of the heels of a plurality of subjects and tested for its effectiveness, it cracked after 2 weeks. It was confirmed that the part was healed in half and was fully replenished after 5 weeks.
本発明の組成の特性と効果の説明
説明１（糖セラミド類の検出）
原料として脱脂大豆を用い、麹菌としてアスペルギルス・オリゼーを用いて前記の生産工程に従って本発明の皮膚外用剤に用いる固形生成物を春夏秋冬の各季節において生産して製品春、製品夏、製品秋、製品冬とした。これらの製品に対して、高速液体クロマトグラフ質量分析法により糖セラミド類（大豆糖セラミドを指標とする）の含有量を測定したところ、０．５５ｍｇ／ｇの割合で含有されていることがわかった。
Description of characteristics and effects of composition of the present invention Description 1 (Detection of sugar ceramides)
Solid products used in the topical skin preparation of the present invention according to the above production process using defatted soybeans as raw materials and Aspergillus oryzae as koji molds are produced in each season of spring, summer, autumn and winter. The product was winter. For these products, the content of sugar ceramides (using soy sugar ceramide as an index) was measured by high performance liquid chromatography / mass spectrometry and found to be contained at a rate of 0.55 mg / g. It was.
この糖セラミド類が含有されることにより、本発明の皮膚外用剤（化粧品を含む）には肌の保湿作用に優れたものとなることがわかった。 It has been found that the inclusion of these sugar ceramides gives the skin external preparation (including cosmetics) of the present invention an excellent skin moisturizing action.
説明２（固形生成物のｐＨの検出）
前記実施例１に用いた固形生成物のｐＨを測定したところ、弱酸性（ｐＨ５．０〜６．０）であった。
この固形生成物が弱酸性を呈することにより、本発明の皮膚外用剤を肌に塗ると、肌の常在菌（例えば、表皮ブドウ球菌）のバランスを整えることとなり、肌の表面のバリア機能を向上させて、肌の健全性を高めることがわかった。
Explanation 2 (Detection of pH of solid product)
When the pH of the solid product used in Example 1 was measured, it was weakly acidic (pH 5.0 to 6.0).
When this solid product exhibits weak acidity, when the external preparation for skin of the present invention is applied to the skin, the balance of normal bacteria of the skin (for example, Staphylococcus epidermidis) will be adjusted, and the barrier function of the skin surface will be improved. It has been found that it improves and improves skin health.
説明３（固形生成物の麹多糖類によるＴｈ２サイトカインへの耐性）
前記実施例１に用いた固形生成物のＴｈ２サイトカインへの耐性を検証するためにＤ１０．Ｇ４．１（Ｄ１０）細胞を利用して確認した。
このＤ１０細胞は、ＡＫＲマウス由来のコンアルブミン（ＣＡ：卵白アレルゲン）特異的Ｔｈ２細胞クローンであり、花粉症、喘息、アトピー性皮膚炎、アナフィラキシーショック等のＩｇＥ誘導性アレルギーにおけるメカニズム検討に利用されているものである。
細胞株・微生物株・遺伝子バンク（ＡＴＣＣ：米国）よりＤ１０細胞（ＴＩＢ２２４）を得て、ＡＫＲマウスＢ細胞（脾臓の細胞）と混合培養することによりＤ１０細胞の継体を得た。
５０万Ｄ１０細胞を３組用意し、前記固形生成物の最終濃度を１組：０％、２組：５０ｎｇ／ｍｌ、３組：２００ｎｇ／ｍｌを投与し、各組とも１００μｇ／ｍｌのＣＡ（卵白アレルゲン）を１回与えて刺激し、気温３７℃、５％ＣＯ２雰囲気７２時間培養後、上澄を採取して、各種のＴｈ２サイトカイン（１組：ＩＬ−４、２組：ＩＬ−５、３組：ＩＬ−１３）をＥＬＩＳＡ法によって測定した。
その測定結果は図２に示す通りとなり、前記固形生成物を投与した２組および３組が、投与しない１組に比較して、投与量が多くなるに従ってＴｈ２細胞サイトカイン（ＩＬ−４、ＩＬ−５、ＩＬ−１３）の生産をより大きく抑制することができることが確認された。
この固形生成物がＴｈ２サイトカインへの耐性を呈することにより、本発明の皮膚外用剤を肌に塗ると、固形生成物中の前記麹多糖類の機能によってアトピー性皮膚炎の軽減や治癒がなされて、肌の健全性を高めることがわかった。
Explanation 3 (Tolerance to Th2 cytokines by cocoon polysaccharide in solid product)
In order to verify the resistance of the solid product used in Example 1 to Th2 cytokine, D10. This was confirmed using G4.1 (D10) cells.
This D10 cell is a Con2 albumin (CA: egg white allergen) -specific Th2 cell clone derived from an AKR mouse and is used for studying mechanisms in IgE-induced allergies such as hay fever, asthma, atopic dermatitis, and anaphylactic shock. It is what.
D10 cells (TIB224) were obtained from cell lines, microorganism strains, and gene banks (ATCC: USA), and D10 cell subcultures were obtained by co-culturing with AKR mouse B cells (spleen cells).
Three sets of 500,000 D10 cells were prepared, and the final concentration of the solid product was administered as 1 set: 0%, 2 sets: 50 ng / ml, 3 sets: 200 ng / ml, and each set was administered with 100 μg / ml CA ( Egg white allergen) was stimulated once, cultured at a temperature of 37 ° C. and 5% CO 2 atmosphere for 72 hours, and the supernatant was collected and various Th2 cytokines (1 set: IL-4, 2 sets: IL-5). Three sets: IL-13) were measured by ELISA.
The measurement results are as shown in FIG. 2, and the Th2 cell cytokines (IL-4, IL-) were increased as the dose increased in 2 and 3 groups administered with the solid product compared to 1 group not administered. 5. It was confirmed that the production of IL-13) can be greatly suppressed.
When this solid product exhibits resistance to Th2 cytokines, when the external preparation for skin of the present invention is applied to the skin, atopic dermatitis is reduced or cured by the function of the cocoon polysaccharide in the solid product. , Found to enhance skin health.
説明４（大腸菌およびブドウ球菌への耐性の検出）
前記実施例１に用いた固形生成物に対する抗菌力試験を日本食品分析センターに依頼して下記の結果を得た。
Ａ 試験方法
１）試験菌
Escherichia coli NBRC 3972（大腸菌）
Stphylicoccus aureus subsp. aureus NBRC 12732（黄色ブドウ球菌）
２）菌数測定用培地及び培養条件
SCDLP 寒天培地（日本製薬株式会社）、混釈平板培養法、３５℃±１℃、２日間
３）試験菌液の調整
試験菌を普通ブイヨン培地（栄研化学株式会社）で３５℃±１℃、１８〜２４時間培養した後、普通ブイヨン培地を用いて、菌数が１０４〜１０５／ｍＬとなるように調整し、試験菌数とした。
４）試験操作
検体１０ｍＬに試験菌液１ｍＬを接種し、試験液とした。試験液を３５℃±１℃で保存し、２４時間後に試験液をSCDLP 寒天培地（日本製薬株式会社）で直ちに１０倍に希釈し、試験液中の生菌数を菌数測定用培地を用いて測定した。
なお、対照としてリン酸緩衝生理食塩水を用いて同様に試験し、開始時についても生菌数を測定した。
Explanation 4 (Detection of resistance to E. coli and staphylococci)
An antibacterial activity test for the solid product used in Example 1 was requested to the Japan Food Analysis Center, and the following results were obtained.
A Test method 1) Test bacteria
Escherichia coli NBRC 3972
Stphylicoccus aureus subsp. Aureus NBRC 12732 (Staphylococcus aureus)
2) Culture medium and culture conditions for bacterial count measurement
SCDLP agar medium (Nippon Pharmaceutical Co., Ltd.), pour plate culture method, 35 ° C ± 1 ° C, 2 days 3) Preparation of test bacterial solution 35 ° C ± 1 ° C in normal broth medium (Eiken Chemical Co., Ltd.) After culturing for 18 to 24 hours, the number of bacteria was adjusted to 10 4 to 10 5 / mL using a normal bouillon medium to obtain the number of test bacteria.
4) Test operation 10 mL of a sample was inoculated with 1 mL of a test bacterial solution to prepare a test solution. The test solution is stored at 35 ° C. ± 1 ° C., and after 24 hours, the test solution is immediately diluted 10-fold with SCDLP agar medium (Nippon Pharmaceutical Co., Ltd.), and the number of viable bacteria in the test solution is measured using a medium for measuring the number of bacteria Measured.
In addition, it tested similarly using the phosphate buffered saline as a control, and the viable cell count was measured also at the start.
Ｂ 試験結果
試験結果は表２に示す通りとなった。
B Test results The test results are shown in Table 2.
この抗菌性試験結果より、２４時間の大腸菌増殖は対照に対してイムバランス抽出液を与えると６／１０００に抑えられることがわかった。
この抗菌性試験結果より、２４時間の黄色ブドウ球菌増殖は対照に対してイムバランス抽出液を与えると１／２００００に抑えられることがわかった。
この固形生成物が黄色ブドウ球菌および大腸菌の増殖を抑制する機能を呈することにより、固形生成物中に含まれるペプチドにバクテリオシンが含まれることがわかった。
From this antibacterial test result, it was found that the growth of E. coli for 24 hours was suppressed to 6/1000 when the imbalance extract was given to the control.
From this antibacterial test result, it was found that the growth of S. aureus for 24 hours was suppressed to 1/20000 when the imbalance extract was given to the control.
It was found that the solid product contained a bacteriocin in the peptide contained in the solid product by exhibiting a function of suppressing the growth of Staphylococcus aureus and Escherichia coli.
このことより本発明の固形生成物は悪玉菌に対する天然抗菌物として作用することがわかり、肌の湿疹や皮膚炎症を抑えることができることがわかった。 From this, it was found that the solid product of the present invention acts as a natural antibacterial product against bad bacteria, and it was found that skin eczema and skin inflammation can be suppressed.
なお、本発明は、前述した実施の形態に限定されるものではなく、必要に応じて種々の変更が可能である。 In addition, this invention is not limited to embodiment mentioned above, A various change is possible as needed.
Claims (7)
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| JP2017193495A (en) * | 2016-04-18 | 2017-10-26 | 株式会社創研 | Balance adjusting agents for skin-indigenous bacteria |
| CN111607524A (en) * | 2019-02-22 | 2020-09-01 | 科丝美诗株式会社 | Aspergillus oryzae strain and composition containing fermentation product thereof for improving skin condition |
| WO2024090464A1 (en) * | 2022-10-25 | 2024-05-02 | 株式会社アインホールディングス | Skin improvement agent having fermented extract as main raw material |
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| JP2009114095A (en) * | 2007-11-02 | 2009-05-28 | Nichimo Co Ltd | Raw preparation for ameliorating allergic symptom |
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| JP2012228246A (en) * | 2011-04-15 | 2012-11-22 | Saga Univ | Method of manufacturing sphingolipid |
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| JP2000264832A (en) * | 1999-03-18 | 2000-09-26 | Fancl Corp | Skin cosmetic |
| JP2009114095A (en) * | 2007-11-02 | 2009-05-28 | Nichimo Co Ltd | Raw preparation for ameliorating allergic symptom |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2017193495A (en) * | 2016-04-18 | 2017-10-26 | 株式会社創研 | Balance adjusting agents for skin-indigenous bacteria |
| CN111607524A (en) * | 2019-02-22 | 2020-09-01 | 科丝美诗株式会社 | Aspergillus oryzae strain and composition containing fermentation product thereof for improving skin condition |
| CN111607524B (en) * | 2019-02-22 | 2024-03-26 | 科丝美诗株式会社 | Aspergillus oryzae strain and composition containing fermentation product thereof for improving skin state |
| WO2024090464A1 (en) * | 2022-10-25 | 2024-05-02 | 株式会社アインホールディングス | Skin improvement agent having fermented extract as main raw material |
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