JP2015159806A - Banana enzyme-treated product and its utilization - Google Patents
Banana enzyme-treated product and its utilization Download PDFInfo
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本発明は、バナナ酵素処理物およびその利用に関するものである。 The present invention relates to a banana enzyme-treated product and use thereof.
フィリピン等の原産国で栽培されたバナナは青くて硬い未熟な状態で収穫される。未熟なバナナは、果肉が硬く、未調理では食用には適さないことが知られている。収穫されたバナナは、傷や形がチェックされて選別され、傷のあるバナナや形のよくないバナナは流通から外れる。未収穫のバナナ（木に取り残されたバナナ）も流通から外れる。選別されて消費国に輸入されたバナナは、各国の店頭に並ぶ前に品質検査を受け、検査に合格しなかったバナナ（未熟なバナナ、成熟バナナを含む。）は商品価値がないものとして廃棄されている。このようにして、最終消費者に行き渡るまでの流通過程おいて、原産国および消費国で処分されているバナナが多数存在する。 Bananas grown in the country of origin such as the Philippines are harvested in a blue, hard and immature state. It is known that immature bananas have a hard pulp and are not suitable for consumption if uncooked. Harvested bananas are checked for scratches and shapes and sorted, and damaged and poorly shaped bananas are out of circulation. Unharvested bananas (bananas left on the tree) are also out of circulation. Bananas that have been sorted and imported into the consuming country undergo quality inspection before they are placed in stores in each country, and bananas that do not pass the inspection (including immature bananas and mature bananas) are discarded as having no commercial value. Has been. In this way, there are many bananas that are disposed of in the country of origin and consumer in the distribution process up to the final consumer.
本発明は、原産国において流通から外れたバナナ、消費国において廃棄されていたバナナ（未熟なバナナ、成熟バナナを含む。）を有効利用することを課題とする。 It is an object of the present invention to effectively utilize bananas that are out of circulation in the country of origin and bananas that are discarded in the consuming country (including immature bananas and mature bananas).
本発明者らは、上記課題を解決するために鋭意研究を重ねた結果、バナナの乾燥粉末を酵素処理することによって、バナナが免疫増強作用と有することを見出し、この知見に基づいてさらに研究を進め、本発明を完成するに至った。 As a result of intensive studies to solve the above problems, the present inventors have found that banana has an immunopotentiating effect by treating banana dry powder with an enzyme, and further research based on this finding. The present invention has been completed.
本発明は、上記課題を解決するために、以下の各発明を包含する。
［１］乾燥バナナを酵素処理することにより調製されることを特徴とするバナナ酵素処理物。
［２］乾燥バナナが、未熟なバナナを乾燥し粉砕したものであることを特徴とする［１］に記載のバナナ酵素処理物。
［３］酵素が、糖質分解酵素である［１］または［２］に記載のバナナ酵素処理物。
［４］酵素が、糖質分解酵素および蛋白質分解酵素である［１］または［２］に記載のバナナ酵素処理物。
［５］糖質分解酵素が、α−アミラーゼ、β−アミラーゼ、β−グルコシダーゼ、セルラーゼ、プルラナーゼ、ペクチナーゼ、ペクトリアーゼ、グルコアミラーゼ、アミログルコシダーゼ、デキストラナーゼ、ヘミセルラーゼ、グルカナーゼ、キシラナーゼおよびこれらの組み合わせからなる群より選択されることを特徴とする［３］または［４］に記載のバナナ酵素処理物。
［６］酵素処理前に、乾燥バナナをアルファ化処理することを特徴とする［１］〜［５］のいずれかに記載のバナナ酵素処理物。
［７］酵素処理後に、酵素処理物を発酵させることを特徴とする［１］〜［６］のいずれかに記載のバナナ酵素処理物。
［８］発酵が、酵母、乳酸菌、納豆菌、麹菌、糸状菌および放線菌からなる群より選択される少なくとも１種の微生物を用いて行われることを特徴とする［７］に記載のバナナ酵素処理物。
［９］水溶性粉末である［１］〜［８］のいずれかに記載のバナナ酵素処理物。
［１０］［１］〜［９］のいずれかに記載のバナナ酵素処理物を含有することを特徴とする飲食品。
［１１］［１］〜［９］のいずれかに記載のバナナ酵素処理物を含有することを特徴とするサリメント。
［１２］［１］〜［９］のいずれかに記載のバナナ酵素処理物を含有することを特徴とする免疫増強剤。
［１３］［１］〜［９］のいずれかに記載のバナナ酵素処理物を含有することを特徴とするミトコンドリア活性化促進剤。
［１４］［１］〜［９］のいずれかに記載のバナナ酵素処理物の製造方法であって、乾燥バナナを酵素処理する酵素処理工程を含むことを特徴とする製造方法。
［１５］酵素処理工程の前に未熟なバナナから乾燥バナナを調製する乾燥バナナ調製工程を含むことを特徴とする［１４］に記載の製造方法。
The present invention includes the following inventions in order to solve the above problems.
[1] A banana enzyme-treated product prepared by subjecting a dried banana to an enzyme treatment.
[2] The banana enzyme-treated product according to [1], wherein the dried banana is obtained by drying and pulverizing an immature banana.
[3] The banana enzyme-treated product according to [1] or [2], wherein the enzyme is a saccharide-degrading enzyme.
[4] The banana enzyme-treated product according to [1] or [2], wherein the enzyme is a saccharide-degrading enzyme or a proteolytic enzyme.
[5] The carbohydrase is α-amylase, β-amylase, β-glucosidase, cellulase, pullulanase, pectinase, pectinase, glucoamylase, amyloglucosidase, dextranase, hemicellulase, glucanase, xylanase and combinations thereof. The banana enzyme-treated product according to [3] or [4], which is selected from the group consisting of:
[6] The banana enzyme-treated product according to any one of [1] to [5], wherein the dried banana is pregelatinized before the enzyme treatment.
[7] The banana enzyme-treated product according to any one of [1] to [6], wherein the enzyme-treated product is fermented after the enzyme treatment.
[8] The banana enzyme according to [7], wherein the fermentation is performed using at least one microorganism selected from the group consisting of yeast, lactic acid bacteria, natto bacteria, koji molds, filamentous fungi, and actinomycetes Processed product.
[9] The banana enzyme-treated product according to any one of [1] to [8], which is a water-soluble powder.
[10] A food or drink comprising the banana enzyme-treated product according to any one of [1] to [9].
[11] A saliment comprising the banana enzyme-treated product according to any one of [1] to [9].
[12] An immune enhancer comprising the banana enzyme-treated product according to any one of [1] to [9].
[13] A mitochondria activation promoter comprising the banana enzyme-treated product according to any one of [1] to [9].
[14] A method for producing the banana enzyme-treated product according to any one of [1] to [9], comprising an enzyme treatment step of enzymatically treating a dried banana.
[15] The production method according to [14], including a dry banana preparation step of preparing a dry banana from an immature banana before the enzyme treatment step.
本発明によれば、原産国において流通から外れたバナナ、消費国において廃棄されていたバナナ（未熟なバナナ、成熟バナナを含む。）を有効利用することができる。本発明によれば、甘みを呈するバナナ酵素処理物を提供することができる。本発明のバナナ酵素処理物は、水溶性を有する。さらに、本発明のバナナ酵素処理物は、免疫増強剤、ミトコンドリア活性化促進剤等として利用することができる。本発明のバナナ酵素処理物は、飲食品、サプリメント、飼料等の成分として利用することができる。本発明のバナナ酵素処理物は、酵素処理後に発酵させることによりさらに高い免疫増強作用を有するという、当業者に予想外の、本発明特有の特に顕著な効果を有する。 According to the present invention, it is possible to effectively utilize bananas that are out of circulation in the country of origin and bananas that are discarded in the consuming country (including immature bananas and mature bananas). ADVANTAGE OF THE INVENTION According to this invention, the banana enzyme processed material which exhibits sweetness can be provided. The banana enzyme-treated product of the present invention has water solubility. Furthermore, the banana enzyme-treated product of the present invention can be used as an immune enhancer, a mitochondrial activation promoter and the like. The banana enzyme-treated product of the present invention can be used as a component for foods and drinks, supplements, feeds and the like. The banana enzyme-treated product of the present invention has a particularly remarkable effect peculiar to the present invention, which is unexpected to those skilled in the art, and has a higher immune enhancing effect by being fermented after the enzyme treatment.
本発明は、乾燥バナナを酵素処理することにより調製されるバナナ酵素処理物（以下、「本発明のバナナ酵素処理物」という。）を提供する。 The present invention provides a banana enzyme-treated product (hereinafter referred to as “the banana enzyme-treated product of the present invention”) prepared by enzymatic treatment of dried banana.
〔乾燥バナナ〕
乾燥バナナの原料であるバナナ（学名：Musa spp.）はバショウ科バショウ属の草本植物の果実の総称である。バナナは殆どが食品用に栽培されているが、繊維用または観賞用にも栽培されている。バナナの種類は特に限定されないが、例えばグロスミッチェル、キャベンディッシュ、プランテイン等が挙げられる。
[Dried banana]
Banana (scientific name: Musa spp.), The raw material of dried bananas, is a generic term for the fruits of herbaceous plants belonging to the genus Baxaceae. Bananas are mostly cultivated for food, but are also cultivated for fiber or ornamental purposes. Although the kind of banana is not specifically limited, For example, gross mitchell, cavendish, plantain, etc. are mentioned.
乾燥バナナの原料に用いるバナナの部位は特に限定されないが、全果実（果実の食用部分と皮を含む）が好ましく、皮を除いた果実の食用部分がより好ましい。 Although the part of the banana used for the raw material of the dried banana is not particularly limited, the whole fruit (including the edible part of the fruit and the skin) is preferable, and the edible part of the fruit excluding the skin is more preferable.
バナナは、まだ青くて硬いうちに収穫され、消費国に輸出される。例えば、日本に輸入されたバナナは、室（むろ）と呼ばれるところで追熟され、黄色くなる。追熟は、バナナの場合、温度、湿度をコントロールされた室で、少量のエチレンを与えながら行われる。乾燥バナナの原料に用いるバナナの成熟度合いは特に限定されず、未熟な青バナナおよび成熟した黄バナナのいずれも好適に用いることができる。乾燥のしやすさ、コスト等の点で未熟な青バナナが好ましい。ここで、未熟な青バナナとは追熟していないバナナを意味し、収穫時期に至っていないバナナ、収穫時期のバナナ、輸入直後のバナナ等の未調理では食用に適さないバナナが含まれる。 Bananas are harvested while still blue and hard and are exported to the consuming country. For example, a banana imported to Japan is ripened in a place called Muro and turns yellow. In the case of bananas, ripening is performed in a room where the temperature and humidity are controlled while supplying a small amount of ethylene. The maturity degree of the banana used for the raw material of the dried banana is not particularly limited, and any of immature green banana and mature yellow banana can be suitably used. An immature green banana is preferable in terms of ease of drying and cost. Here, an immature green banana means a banana that has not yet been ripened, and includes bananas that are not edible for cooking, such as bananas that have not yet been harvested, bananas that have been harvested, and bananas that have just been imported.
乾燥バナナの調製方法は、例えばバナナを熱風乾燥、自然乾燥、乾熱乾燥等する方法が挙げられる。乾燥には公知の乾燥装置を使用することができる。乾燥後さらに粉砕等してもよい。粉砕には公知の粉砕装置を使用することができる。乾燥バナナは、乾燥バナナ全量に対して水分量が２０重量％以下であることが好ましい。乾燥バナナの形態は特に限定されず、バナナの形状そのまま、チップ状、粒状、粉末状等が挙げられる。乾燥バナナは市販品を入手することも可能である。市販品として例えば、バナナチップ、Ｔｏｏｋｅ（ウガンダ産）プランテイン、バナナ粉（Ｄｏｌｅ製）等が挙げられる。 Examples of the method for preparing the dried banana include a method of drying the banana with hot air, natural drying, and dry heat drying. A known drying apparatus can be used for drying. You may grind | pulverize further after drying. For the pulverization, a known pulverizer can be used. The dried banana preferably has a water content of 20% by weight or less based on the total amount of the dried banana. The form of the dried banana is not particularly limited, and examples of the shape of the banana include a chip shape, a granular shape, and a powder shape. Commercially available dried bananas can also be obtained. Examples of commercially available products include banana chips, Tooké (Uganda) plantain, banana powder (manufactured by Dole), and the like.
〔酵素処理〕
酵素は糖質を分解可能な酵素を含むことが好ましい。糖質分解酵素として、例えばα−アミラーゼ、β−アミラーゼ、β−グルコシダーゼ、セルラーゼ、プルラナーゼ、ペクチナーゼ、ペクトリアーゼ、グルコアミラーゼ、アミログルコシダーゼ、デキストラナーゼ、ヘミセルラーゼ、グルカナーゼ、キシラナーゼ等が挙げられ、これらのうちの１種を単独でまたは２種以上を組み合わせて使用することができる。中でもα−アミラーゼが好ましい。酵素として、糖質分解酵素の他に、蛋白質分解酵素、植物組織崩壊酵素、リン脂質分解酵素を含むことが好ましい。蛋白質分解酵素（プロテアーゼ）として、例えば、ペプシン、パパイン、カスパーゼ等が挙げられる。植物組織崩壊酵素として、例えばセルラーゼ、ペクチナーゼ等が挙げられる。リン脂質分解酵素として例えばホスホリパーゼ等が挙げられる。これらのうちの１種を単独でまたは２種以上を組み合わせて使用することができる。
[Enzyme treatment]
The enzyme preferably contains an enzyme capable of degrading carbohydrates. Examples of the carbohydrase include α-amylase, β-amylase, β-glucosidase, cellulase, pullulanase, pectinase, pectinase, glucoamylase, amyloglucosidase, dextranase, hemicellulase, glucanase, xylanase, and the like. One of them can be used alone or in combination of two or more. Of these, α-amylase is preferred. In addition to the saccharide-degrading enzyme, the enzyme preferably contains a proteolytic enzyme, a plant tissue-degrading enzyme, and a phospholipid degrading enzyme. Examples of proteolytic enzymes (proteases) include pepsin, papain, caspase and the like. Examples of plant tissue disrupting enzymes include cellulase and pectinase. Examples of the phospholipid degrading enzyme include phospholipase. One of these can be used alone or in combination of two or more.
これらの酵素は、例えば液化酵素６Ｔ（エイチビィアイ製）、液化酵素Ｔ（エイチビィアイ製）、ビオザイムＡ（天野エンザイム）、ペクチナーゼ“ナガセ”（ナガセケムテックス）、ヘミセルラーゼ「アマノ」90（天野エンザイム）、β-アミラーゼＦ（天野エンザム）の市販品を用いることができる。 These enzymes include, for example, liquefaction enzyme 6T (manufactured by HIBI), liquefaction enzyme T (manufactured by HIBI), biozyme A (Amano Enzyme), pectinase “Nagase” (Nagase ChemteX), hemicellulase “Amano” 90 (Amano Enzyme), A commercially available product of β-amylase F (Amano Enzam) can be used.
酵素処理の方法は、例えば乾燥バナナに水を加えた液を攪拌し、使用する酵素の至適温度、至適ｐＨに調整し、酵素を添加する方法等が挙げられる。水の量は特に限定されないが、例えば乾燥バナナの重量に対して５〜１５倍の重量が好ましく、７〜１３倍の重量がより好ましく、９〜１０倍の重量がさらに好ましい。ｐＨは水酸化ナトリウム、塩酸等を使用して調整しうる。 Examples of the enzyme treatment method include a method in which a solution obtained by adding water to dried banana is stirred, adjusted to the optimum temperature and pH of the enzyme to be used, and the enzyme is added. The amount of water is not particularly limited, but is preferably 5 to 15 times the weight of the dried banana, more preferably 7 to 13 times the weight, and even more preferably 9 to 10 times the weight. The pH can be adjusted using sodium hydroxide, hydrochloric acid or the like.
酵素の添加量は乾燥バナナを可溶化できる量であればよく、用いる乾燥バナナおよび用いる酵素によって至適添加量が異なるため、その都度設定することが好ましい。通常、乾燥バナナを含有する水の全量に対して０．０１〜５．０重量％が用いられる。
酵素処理工程により、本発明のバナナ酵素処理物が水溶性を獲得し、また甘味を呈するという効果が得られる。
The amount of the enzyme added is not particularly limited as long as it can solubilize the dried banana, and the optimum amount of addition varies depending on the dried banana to be used and the enzyme to be used. Usually, 0.01 to 5.0% by weight is used with respect to the total amount of water containing dried bananas.
By the enzyme treatment step, an effect that the banana enzyme-treated product of the present invention acquires water solubility and exhibits sweetness can be obtained.
〔アルファ化処理〕
本発明のバナナ酵素処理物は、乾燥バナナをアルファ化処理して製造することが好ましい。アルファ化処理は酵素処理と同時に行ってもよく酵素処理の前に行ってもよいが、酵素処理の前に行うことが好ましい。
[Alpha processing]
The banana enzyme-treated product of the present invention is preferably produced by subjecting a dried banana to pregelatinization. The pregelatinization treatment may be performed simultaneously with the enzyme treatment or may be performed before the enzyme treatment, but is preferably performed before the enzyme treatment.
アルファ化処理の方法は特に限定されないが、例えば乾燥バナナに水を加えて加熱する方法、水を加えて加圧する方法等が挙げられる。水の量は特に限定されないが、例えば乾燥バナナの重量に対して５〜１５倍の重量が好ましく、７〜１３倍の重量がより好ましく、９〜１０倍の重量がさらに好ましい。加熱によりアルファ化処理を行う場合、加熱温度は特に限定されないが、例えば５０〜１５０℃であることが好ましく、６０〜１３０℃であることがより好ましく、７５〜１２５℃であることがさらに好ましい。加熱時間はアルファ化処理の効果が妨げられない限り特に限定されず、加熱温度に応じて設定することも可能である。加熱時間として例えば１〜１８０分間が好ましく、５〜１２０分間がより好ましい。加熱温度および加熱時間の好ましい組み合わせとして例えば７５℃で２０〜１８０分間、８０℃で２０〜１２０分間、１２１℃で５〜３０分等が挙げられる。 The method of the pregelatinization treatment is not particularly limited, and examples thereof include a method in which water is added to a dried banana and heated, and a method in which water is added and pressurized. The amount of water is not particularly limited, but is preferably 5 to 15 times the weight of the dried banana, more preferably 7 to 13 times the weight, and even more preferably 9 to 10 times the weight. When the pregelatinization treatment is carried out by heating, the heating temperature is not particularly limited, but is preferably, for example, 50 to 150 ° C, more preferably 60 to 130 ° C, and further preferably 75 to 125 ° C. The heating time is not particularly limited as long as the effect of the pregelatinization treatment is not hindered, and can be set according to the heating temperature. For example, the heating time is preferably 1 to 180 minutes, more preferably 5 to 120 minutes. Preferred combinations of heating temperature and heating time include, for example, 75 ° C. for 20 to 180 minutes, 80 ° C. for 20 to 120 minutes, 121 ° C. for 5 to 30 minutes, and the like.
上記酵素処理、または上記アルファ化処理および上記酵素処理により、目的の酵素処理物が得られる。酵素処理後に、得られた酵素処理物をそのまま酵素処理物として使用してもよく、一旦保存して必要に応じて使用してもよい。また、得られた酵素処理物は後述の発酵処理の材料として使用してもよい。 The target enzyme-treated product is obtained by the enzyme treatment, or the pregelatinization treatment and the enzyme treatment. After the enzyme treatment, the obtained enzyme-treated product may be used as it is as an enzyme-treated product, or may be stored once and used as necessary. Moreover, you may use the obtained enzyme processed material as a material of the below-mentioned fermentation process.
〔発酵処理〕
本発明のバナナ酵素処理物は、乾燥バナナを発酵処理して製造することが好ましい。発酵処理は、酵素処理の後に行うことが好ましい。
[Fermentation treatment]
The banana enzyme-treated product of the present invention is preferably produced by fermenting a dried banana. The fermentation treatment is preferably performed after the enzyme treatment.
発酵処理に用いる微生物は特に限定されないが、例えば酵母、乳酸菌、納豆菌、麹菌、糸状菌、放線菌等が挙げられ、これらのうちの１種を単独でまたは２種以上を組み合わせて使用することができる。酵母としては、例えば、サッカロミセス属、シゾサッカロミセス属、カンジダ属、クルイベロミセス属等が挙げられる。これらの中でも、飲食品用途の観点から、サッカロミセス属の酵母が好ましい。乳酸菌としては、例えば、ラクトバチラス属、ラクトコッカス属、ペディオコッカス属、ストレプトコッカス属、エンテロコッカス属、ロイコノストック属、テトラゲノコッカス属、エノコッカス属、およびワイセラ属等が挙げられる。納豆菌としては、バチラス属等が挙げられる。麹菌としては黄麹菌、白麹菌等が挙げられる。糸状菌としては、例えば、アクレモニウム属、アスペルギルス属、セリポリオプシス属、ケトミウム属、コプリナス属、ジオトリチューム属、ヒュミコラ属、モナスカス属、ムコール属、ペニシリウム属、ファネロケート属、リゾムコール属、リゾプス属、トラメテス属、およびトリコデルマ属が挙げられる。放線菌としては、例えば、アクチノプラネス属、アミコラトプシス属、ストレプトマイセス属、ストレプトバーチシリウム属、およびサーモモノスポラ属が挙げられる。 The microorganism used for the fermentation treatment is not particularly limited, and examples thereof include yeast, lactic acid bacteria, natto bacteria, koji molds, filamentous fungi, actinomycetes, etc., and one of these may be used alone or in combination of two or more. Can do. Examples of yeasts include Saccharomyces, Schizosaccharomyces, Candida, Kluyveromyces and the like. Among these, yeast of the genus Saccharomyces is preferable from the viewpoint of food and drink use. Examples of the lactic acid bacteria include Lactobacillus, Lactococcus, Pediococcus, Streptococcus, Enterococcus, Leuconostoc, Tetragenococcus, Enococcus, and Weicella. Examples of Bacillus natto include Bacillus. Examples of the koji mold include yellow koji mold, white koji mold, and the like. Examples of the filamentous fungi include, for example, Acremonium, Aspergillus, Seripoliopsis, Ketomium, Coprinas, Geotritum, Humicola, Monascus, Mucor, Penicillium, Phanerocate, Rhizomucor, Rhizopus , Trametes, and Trichoderma. Examples of actinomycetes include the genus Actinoplanes, the genus Amycolatopsis, the genus Streptomyces, the genus Streptomyces and the genus Thermomonospora.
上記発酵は、酵素処理物と、微生物とを接触させることにより行われる。接触方法については特に制限されない。例えば、酵素処理物を含む液中に、微生物の種培養液を添加する方法が挙げられる。発酵条件についても特に制限されず、発酵温度および発酵時間は、添加する種菌の量、微生物の種類に応じて適宜設定される。例えば、微生物として酵母を用いる場合、発酵ｐＨは４〜９であることが好ましく、５〜８であることがより好ましく、６〜７であることがさらに好ましい。微生物として酵母を用いる場合、発酵温度は２０〜５０℃であることが好ましく、２５〜４５℃であることがより好ましく、３０〜４０℃であることがさらに好ましい。微生物として酵母を用いる場合、発酵時間は例えば１〜１２時間であることが好ましく、２〜６時間であることがより好ましく、２〜５時間がさらに好ましい。
なお、本発明は、酵素処理を行わずに乾燥バナナをアルファ化処理後発酵させることによっても得られる乾燥バナナ処理物も含む。
The fermentation is performed by bringing the enzyme-treated product into contact with a microorganism. The contact method is not particularly limited. For example, a method of adding a seed culture solution of a microorganism to a solution containing an enzyme-treated product can be mentioned. The fermentation conditions are not particularly limited, and the fermentation temperature and fermentation time are appropriately set according to the amount of inoculum to be added and the type of microorganism. For example, when yeast is used as the microorganism, the fermentation pH is preferably 4 to 9, more preferably 5 to 8, and still more preferably 6 to 7. When yeast is used as the microorganism, the fermentation temperature is preferably 20 to 50 ° C, more preferably 25 to 45 ° C, and further preferably 30 to 40 ° C. When using yeast as a microorganism, the fermentation time is preferably 1 to 12 hours, more preferably 2 to 6 hours, and even more preferably 2 to 5 hours.
In addition, this invention also includes the dried banana processed material obtained also by fermenting a dried banana after pregelatinization without performing an enzyme treatment.
〔固液分離、粉末化〕
本発明のバナナ酵素処理物は、酵素処理により得られる状態のまま使用してもよいし、本発明の効果を奏する限り、例えば酵素処理物を固液分離し残渣を除去する固液分離後の状態で使用してもよいし、さらに固液分離工程で得られた酵素処理物を粉末化する粉末化した状態で使用してもよい。
固液分離は、酵素処理の後に行うことが好ましい。
[Solid-liquid separation, powderization]
The banana enzyme-treated product of the present invention may be used in the state obtained by the enzyme treatment, and as long as the effects of the present invention are exhibited, for example, the enzyme-treated product is subjected to solid-liquid separation and the residue is removed after solid-liquid separation. The enzyme-treated product obtained in the solid-liquid separation step may be used in a powdered state.
Solid-liquid separation is preferably performed after the enzyme treatment.
固液分離は、ろ過、遠心分離、遠心ろ過、サイクロン、フィルタープレス、スクリュープレス、デカンター等を使用して行うことができる。 Solid-liquid separation can be performed using filtration, centrifugation, centrifugal filtration, cyclone, filter press, screw press, decanter, and the like.
粉末化は、熱風乾燥、真空乾燥、凍結乾燥、噴霧乾燥等により行うことができる。 The powderization can be performed by hot air drying, vacuum drying, freeze drying, spray drying, or the like.
〔製造方法〕
本発明は、本発明のバナナ酵素処理物の製造方法（以下、「本発明の製造方法」という。）を提供する。本発明の製造方法において使用される原料（バナナ、酵素、発酵に用いる微生物等）は、本発明のバナナ酵素処理物において使用されるものが好ましい。
〔Production method〕
The present invention provides a method for producing the banana enzyme-treated product of the present invention (hereinafter referred to as “the production method of the present invention”). The raw materials (bananas, enzymes, microorganisms used for fermentation, etc.) used in the production method of the present invention are preferably those used in the banana enzyme-treated product of the present invention.
本発明の製造方法は「乾燥バナナを酵素処理する酵素処理工程」を含んでいればよく、本発明のバナナ酵素処理物を製造することができる限り他の工程を含んでいてもよい。酵素処理工程では、上記本発明のバナナ酵素処理物の説明中の〔酵素処理〕において記載した内容に従って乾燥バナナを酵素処理すればよい。本発明の製造方法は、酵素処理工程の前に、「未熟なバナナから乾燥バナナを調製する乾燥バナナ調製工程」を含むことが好ましい。乾燥バナナ調製工程では、上記本発明のバナナ酵素処理物の説明中の〔乾燥バナナ〕において記載した内容に従って乾燥バナナを調製すればよい。 The production method of the present invention only needs to include an “enzyme treatment step for enzymatic treatment of dried banana”, and may include other steps as long as the banana enzyme-treated product of the present invention can be produced. In the enzyme treatment step, the dried banana may be enzymatically treated according to the contents described in [Enzyme treatment] in the explanation of the banana enzyme-treated product of the present invention. The production method of the present invention preferably includes a “dried banana preparation step of preparing a dried banana from an immature banana” before the enzyme treatment step. In the dried banana preparation step, the dried banana may be prepared according to the contents described in [Dried Banana] in the explanation of the banana enzyme-treated product of the present invention.
本発明の製造方法はさらに、「乾燥バナナをアルファ化処理するアルファ化処理工程」、「酵素処理物を発酵させる発酵処理工程」、「酵素処理物を固液分離し残渣を除去する固液分離工程」、「固液分離工程で得られた酵素処理物を粉末化する粉末化工程」等を含むことが好ましい。これらの工程は、それぞれ、上記本発明のバナナ酵素処理物の説明中の〔アルファ化処理〕、〔発酵処理〕、〔固液分離、粉末化〕において記載した内容に従って行うことができる。アルファ化処理工程は、酵素処理工程の前にまたは乾燥バナナ調製工程と酵素処理工程との間であることが好ましい。固液分離工程は酵素処理工程の後であることが好ましい。発酵処理工程は酵素処理工程の後に、または酵素処理工程と固液分離工程との間であることが好ましい。粉末化工程は固液分離工程の後であることが好ましい。 The production method of the present invention further includes “an alpha-treatment process for alpha-treating dried banana”, “a fermentation treatment process for fermenting an enzyme-treated product”, “solid-liquid separation for solid-liquid separation of the enzyme-treated product and removal of residues It is preferable to include a “step”, a “powdering step of powdering the enzyme-treated product obtained in the solid-liquid separation step” and the like. Each of these steps can be performed according to the contents described in [Alphalation treatment], [Fermentation treatment], and [Solid-liquid separation, pulverization] in the explanation of the banana enzyme-treated product of the present invention. The pregelatinization treatment step is preferably performed before the enzyme treatment step or between the dry banana preparation step and the enzyme treatment step. The solid-liquid separation step is preferably after the enzyme treatment step. The fermentation treatment step is preferably after the enzyme treatment step or between the enzyme treatment step and the solid-liquid separation step. The powdering step is preferably after the solid-liquid separation step.
〔免疫増強剤〕
本発明は、本発明のバナナ酵素処理物を含有する免疫増強剤（以下、本発明の免疫増強剤という。）を提供する。特に、発酵処理工程等を経て製造されたバナナ酵素処理物は強力な免疫増強作用を有する。本発明の免疫増強剤は免疫賦活剤と換言することができる。
[Immune enhancer]
The present invention provides an immunopotentiator containing the banana enzyme-treated product of the present invention (hereinafter referred to as the immunopotentiator of the present invention). In particular, a banana enzyme-treated product produced through a fermentation treatment process or the like has a strong immune enhancing action. The immunopotentiator of the present invention can be restated as an immunostimulator.
〔ミトコンドリア活性化促進剤〕
本発明は、本発明のバナナ酵素処理物を含有するミトコンドリア活性化促進剤（以下、本発明のミトコンドリア活性化促進剤という。）を提供する。特に、発酵処理工程等を経て製造されたバナナ酵素処理物は強力なミトコンドリア活性化促進作用を有する。本発明のミトコンドリア活性化促進剤は活力増強剤と換言することができる。
[Mitochondrial activation promoter]
The present invention provides a mitochondrial activation promoter (hereinafter referred to as the mitochondrial activation promoter of the present invention) containing the banana enzyme-treated product of the present invention. In particular, a banana enzyme-treated product produced through a fermentation treatment process or the like has a strong mitochondrial activation promoting action. The mitochondrial activation promoter of the present invention can be restated as a vitality enhancer.
〔飲食品〕
本発明は、本発明のバナナ酵素処理物を含有する飲食品（以下、本発明の飲食品という。）を提供する。本発明の飲食品は、免疫増強用飲食品、ミトコンドリア活性化促進用飲食品として好適である。本発明の飲食品は、食品でない未熟なバナナを飲食用として利用することができる点で有用である。
[Food and Drink]
This invention provides the food / beverage products (henceforth food / beverage products of this invention) containing the banana enzyme processed material of this invention. The food / beverage products of the present invention are suitable as a food / beverage product for enhancing immunity and a food / beverage product for promoting mitochondrial activation. The food / beverage products of this invention are useful at the point which can utilize the immature banana which is not a foodstuff for food-drinks.
飲食品には、健康食品、機能性食品、特定保健用食品、病者用食品が含まれる。飲食品の形態は特に限定されない。例えば茶飲料、清涼飲料、炭酸飲料、栄養飲料、果実飲料、乳酸飲料等の飲料、そば、うどん、中華麺、即席麺等の麺類、飴、キャンディー、ガム、チョコレート、スナック菓子、ビスケット、ゼリー、ジャム、クリーム、焼き菓子、パン等の菓子およびパン類、かまぼこ、ハム、ソーセージ等の水産・畜産加工食品、加工乳、発酵乳等の乳製品、サラダ油、てんぷら油、マーガリン、マヨネーズ、ショートニング、ホイップクリーム、ドレッシング等の油脂および油脂加工食品、ソース、たれ等の調味料、カレー、シチュー、丼、お粥、雑炊等のレトルトパウチ食品、アイスクリーム、シャーベット、かき氷等の冷菓などを挙げることができる。 The food and drink includes health food, functional food, food for specified health use, and food for the sick. The form of the food or drink is not particularly limited. For example, beverages such as tea beverages, soft drinks, carbonated beverages, nutritive beverages, fruit beverages, lactic acid beverages, buckwheat noodles, Chinese noodles, instant noodles and other noodles, rice cakes, candy, gum, chocolate, snacks, biscuits, jelly, jam , Cream, baked confectionery, confectionery such as bread, and fishery products such as kamaboko, ham, sausage, processed food, dairy products such as processed milk, fermented milk, salad oil, tempura oil, margarine, mayonnaise, shortening, whipped cream And fats and fat processed foods such as dressings, seasonings such as sauces and sauces, retort pouch foods such as curry, stew, rice cakes, rice cakes and miscellaneous foods, and frozen desserts such as ice cream, sherbet and shaved ice.
本発明の飲食品は、本発明のバナナ酵素処理物を妨げないような任意の飲食品に配合したものであってもよいし、本発明のバナナ酵素処理物を主成分とする飲食品であってもよい。 The food / beverage product of the present invention may be blended with any food / beverage product that does not interfere with the banana enzyme-treated product of the present invention, or a food / beverage product containing the banana enzyme-treated product of the present invention as a main component. May be.
〔サプリメント〕
本発明は、本発明のバナナ酵素処理物含有するサプリメント（以下、本発明のサプリメントという。）を含有する。本発明のサプリメントは、免疫増強用サプリメント、ミトコンドリア活性化促進用サプリメントとして好適である。サプリメントは、例えば錠剤、顆粒剤、散剤、ドリンク剤等の形態で提供することができる。
〔supplement〕
The present invention contains a supplement containing the banana enzyme-treated product of the present invention (hereinafter referred to as the supplement of the present invention). The supplement of the present invention is suitable as a supplement for enhancing immunity and a supplement for promoting mitochondrial activation. Supplements can be provided in the form of tablets, granules, powders, drinks, and the like.
本発明のサプリメントを製造する際には、例えば、デキストリン、デンプン等の糖類；ゼラチン、大豆タンパク、トウモロコシタンパク等のタンパク質；アラニン、グルタミン、イソロイシン等のアミノ酸類；セルロース、アラビアゴム等の多糖類；大豆油、中鎖脂肪酸トリグリセリド等の油脂類等の任意の助剤、安定化剤、保存剤、酸化防止剤を添加することができる。 When producing the supplement of the present invention, for example, sugars such as dextrin and starch; proteins such as gelatin, soybean protein and corn protein; amino acids such as alanine, glutamine and isoleucine; polysaccharides such as cellulose and gum arabic; Arbitrary auxiliaries such as soybean oil and fats and oils such as medium-chain fatty acid triglycerides, stabilizers, preservatives and antioxidants can be added.
〔飼料〕
本発明の酵素処理物は、飼料の形態で実施することができる。飼料としては、例えば、ウシ、ウマ、ブタ等の家畜用飼料、ニワトリ等の家禽用飼料、イヌ、ネコ等のペット用飼料などが挙げられる。本発明の飼料は、飼料中に本発明の酵素処理物を添加する以外、一般的な飼料の製造方法を用いて加工製造することができる。
〔feed〕
The enzyme-treated product of the present invention can be implemented in the form of feed. Examples of the feed include feed for livestock such as cattle, horses and pigs, feed for poultry such as chickens, and feed for pets such as dogs and cats. The feed of the present invention can be processed and produced using a general feed production method, except that the enzyme-treated product of the present invention is added to the feed.
本発明には、バナナ酵素処理物の有効量をヒトまたは非ヒト動物に投与する免疫増強方法が含まれる。本発明には、バナナ酵素処理物の有効量をヒトまたは非ヒト動物に投与するミトコンドリア活性化促進方法が含まれる。本発明には、バナナ酵素処理物の有効量をヒトまたは非ヒト動物に投与する非治療的な免疫増強方法も含まれる。本発明には、バナナ酵素処理物の有効量をヒトまたは非ヒト動物に投与する非治療的なミトコンドリア活性化促進方法も含まれる。なお、「非治療的」とは、医療行為、すなわち治療による人体または動物の体への処置行為を含まない概念である。 The present invention includes an immune enhancement method in which an effective amount of a banana enzyme-treated product is administered to a human or non-human animal. The present invention includes a method for promoting mitochondrial activation, in which an effective amount of a banana enzyme-treated product is administered to a human or non-human animal. The present invention also includes a non-therapeutic immune enhancement method in which an effective amount of a banana enzyme-treated product is administered to a human or non-human animal. The present invention also includes a non-therapeutic method for promoting mitochondrial activation in which an effective amount of a banana enzyme-treated product is administered to a human or non-human animal. The term “non-therapeutic” is a concept that does not include a medical act, that is, a treatment act on the human or animal body by treatment.
本発明には、免疫増強剤を製造するためのバナナ酵素処理物の使用が含まれる。本発明には、ミトコンドリア活性化剤を製造するためのバナナ酵素処理物の使用が含まれる。 The present invention includes the use of a banana enzyme-treated product for producing an immune enhancer. The present invention includes the use of a banana enzyme-treated product for producing a mitochondrial activator.
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.
〔１：製造例〕
（１）乾燥バナナ粉末
バナナ粉（ＤＯＬＥ製）を使用した。
（２）加水処理
乾燥バナナ粉末の重量に対して９倍の水を加えて攪拌したが乾燥バナナ粉末は水に溶けなかった。
（３）加水・加熱処理
乾燥バナナ粉末の重量に対して９倍の水を加えて攪拌後、７０℃で１時間加熱したが乾燥バナナ粉末は水に溶けなかった。
[1: Production example]
(1) Dried banana powder Banana powder (made by DOLE) was used.
(2) Hydrolysis treatment Although 9 times as much water as the weight of the dried banana powder was added and stirred, the dried banana powder did not dissolve in water.
(3) Hydrolysis and heat treatment After adding 9 times the water to the weight of the dried banana powder and stirring, the mixture was heated at 70 ° C for 1 hour, but the dried banana powder did not dissolve in water.
（４−１）酵素処理（１）
乾燥バナナ粉末重量に対して９倍の水を加えた後、加熱してα化（糊化）した。その後至適ｐＨ６．３に調整し、乾燥バナナ粉末重量に対して１％の液化酵素６Ｔ（エイチビィアイ製）を添加した。６０℃にて３時間反応させた。
(4-1) Enzyme treatment (1)
After adding 9 times as much water as the dry banana powder weight, it was heated and gelatinized (gelatinized). Thereafter, the pH was adjusted to an optimum pH of 6.3, and 1% of liquefied enzyme 6T (manufactured by HBI) was added to the weight of the dried banana powder. The reaction was carried out at 60 ° C. for 3 hours.
（４−２）酵素処理（２）
（４−１）において、加熱条件を８０℃、３０分間に代えた以外は（４−１）と同様に行った。
(4-2) Enzyme treatment (2)
In (4-1), the procedure was the same as (4-1) except that the heating conditions were changed to 80 ° C. for 30 minutes.
（４−３）酵素処理（３）
（４−１）において、加熱条件を８０℃、１２０分間に代えた以外は（４−１）と同様に行った。
(4-3) Enzyme treatment (3)
In (4-1), the same procedure as in (4-1) was performed except that the heating condition was changed to 80 ° C. for 120 minutes.
（４−４）酵素処理（４）
（４−１）において、加熱条件を１２１℃、１５分間に代えた以外は（４−１）と同様に行った。
(4-4) Enzyme treatment (4)
(4-1) was carried out in the same manner as (4-1) except that the heating conditions were changed to 121 ° C. for 15 minutes.
各処理による可溶化の判断は目視によって行った。酵素処理した乾燥バナナ粉末（（４−１）〜（４−４））は可溶化することに成功した。以下、（４−１）の反応後の溶液を２５℃、８０００ｒｐｍ（１９０×ｇ）で２０分間遠心分離し、遠心上清をスプレードライヤー（入口温度１７０度、出口温度９０度）を用いて粉末化することで得られた粉末を「酵素処理物」と称する。 The judgment of solubilization by each treatment was performed visually. The enzyme-treated dry banana powder ((4-1) to (4-4)) was successfully solubilized. Hereinafter, the solution after the reaction of (4-1) is centrifuged at 25 ° C. and 8000 rpm (190 × g) for 20 minutes, and the centrifuged supernatant is powdered using a spray dryer (inlet temperature 170 °, outlet temperature 90 °). The powder obtained by converting is referred to as “enzyme-treated product”.
〔２：発酵処理〕
（４−１）の反応後の溶液をスーパーカメリア（商品名、日清製粉グループ製）の至適温度３７℃に調整後、溶液全量に対して０．２％のスーパーカメリア加えて、攪拌しながら４時間反応させた。その後０．４５ｎｍフィルターで可溶化成分と不溶成分を分離し、濾液を回収した。濾液を９０℃で２０分間にて酵素・酵母失活処理し、２５℃、８０００ｒｐｍ（１９０×ｇ）で２０分間遠心分離した。遠心上清をスプレードライヤー（入口温度１７０度、出口温度９０度）を用いて熱風乾燥にて粉末化させた。以下、発酵処理後に得られた粉末を「発酵処理物」と称する。
[2: Fermentation treatment]
After adjusting the solution after the reaction of (4-1) to an optimum temperature of 37 ° C. of Super Camellia (trade name, manufactured by Nisshin Flour Milling Group), 0.2% of Super Camellia is added to the total amount of the solution and stirred. The reaction was continued for 4 hours. Thereafter, the solubilized component and the insoluble component were separated by a 0.45 nm filter, and the filtrate was recovered. The filtrate was subjected to enzyme / yeast inactivation treatment at 90 ° C. for 20 minutes, and centrifuged at 25 ° C. and 8000 rpm (190 × g) for 20 minutes. The centrifugal supernatant was pulverized by hot air drying using a spray dryer (inlet temperature 170 °, outlet temperature 90 °). Hereinafter, the powder obtained after the fermentation treatment is referred to as “fermented product”.
〔３：官能試験〕
健康な成人男女１０人をパネラーとして、口腔内に乾燥バナナ粉末、酵素処理物、発酵処理物を含ませ味覚試験を行った。パネラーは、全員乾燥バナナ粉末は苦くてしびれを感じ、酵素処理物及び発酵処理物は甘いと感じた。
[3: Sensory test]
Using 10 healthy adult men and women as panelists, a taste test was conducted with dry banana powder, enzyme-treated products, and fermented products in the mouth. The panelists all felt that the dried banana powder was bitter and numb, and the enzyme-treated product and the fermented product were sweet.
〔４：免疫増強作用（１）〕
本発明の発酵処理物による、マクロファージ様細胞のポリスチレンビーズ貪食能増強効果を検討した。
[4: Immunity enhancing action (1)]
The effect of enhancing the phagocytic activity of polystyrene beads in macrophage-like cells by the fermented product of the present invention was examined.
（１）前培養
マウス由来マクロファージ様細胞株Ｊ７７４．１細胞（ＪＣＲＢ細胞バンク、細胞番号ＪＣＲＢ００１８）は、１０％ＦＢＳ、１００μｇ／ｍＬペニシリンおよび１００μｇ／ｍＬストレプトマイシン含有ＤＭＥＭ培地（以下、培養液Ａという。）にて継代培養したものを用いた。培養にはＴ２５培養フラスコを用い、３または４日毎に１〜２×１０５ｃｅｌｌｓ／ｍＬで植え継いだ。３７℃に設定した５％ＣＯ２インキュベーター内で培養した。
(1) Preculture The mouse-derived macrophage-like cell line J774.1 cells (JCRB cell bank, cell number JCRB0018) is a DMEM medium containing 10% FBS, 100 μg / mL penicillin and 100 μg / mL streptomycin (hereinafter referred to as culture medium A). ) Was used for subculture. T25 culture flasks were used for culture, and the cells were subcultured at 1-2 × 10 5 cells / mL every 3 or 4 days. The cells were cultured in a 5% CO 2 incubator set at 37 ° C.
（２）試験操作
以下の試験操作はクリーンベンチ内で行った。
Ｔ２５培養フラスコにて前培養したＪ７７４．１細胞をピペッティグにより壁から剥がし、得られた細胞の懸濁液を１５ｍＬコニカルチューブに移した。チューブを室温で、１０００ｒｐｍ（１９０×ｇ）８分間、遠心を行い、上清をデカンテーションで捨て、細胞をペレットとして回収した。タッピングにより細胞をほぐした後、培養液を１０ｍＬ加え、室温で、１０００ｒｐｍ（１９０×ｇ）８分間、遠心を行い、上清をデカンテーションで捨て、細胞をペレットとして回収した。細胞ペレットに培養液Ａ５ｍＬを加え、ピペッティングにより細胞を均一に懸濁した。細胞懸濁液１００μＬを１．５ｍＬマイクロチューブに移し、細胞数と生存率を測定した。生存率が９８．３％（９０％以上）であったので、残液を試験に用いた。
(2) Test operation The following test operation was performed in a clean bench.
J774.1 cells pre-cultured in a T25 culture flask were peeled off from the wall by pipetting, and the resulting cell suspension was transferred to a 15 mL conical tube. The tube was centrifuged at 1000 rpm (190 × g) for 8 minutes at room temperature, the supernatant was discarded by decantation, and the cells were collected as a pellet. After loosening the cells by tapping, 10 mL of the culture solution was added, centrifuged at 1000 rpm (190 × g) for 8 minutes at room temperature, the supernatant was discarded by decantation, and the cells were collected as a pellet. 5 mL of the culture medium A was added to the cell pellet, and the cells were uniformly suspended by pipetting. 100 μL of the cell suspension was transferred to a 1.5 mL microtube, and the cell number and viability were measured. Since the survival rate was 98.3% (90% or more), the remaining liquid was used for the test.
測定した細胞数に基づいて、５×１０５ｃｅｌｌｓ／ｍＬの細胞懸濁液を調製した。得られた細胞懸濁液を２００μＬずつカルチャースライドの各ウェルに加えた。３７℃の５％ＣＯ２インキュベーターに移して、細胞がウェルの底に接着して伸展するまで２時間前培養を行った。 Based on the measured cell number, a cell suspension of 5 × 10 5 cells / mL was prepared. 200 μL of the obtained cell suspension was added to each well of the culture slide. The cells were transferred to a 37 ° C. 5% CO 2 incubator and precultured for 2 hours until the cells adhered to the bottom of the well and spread.
各濃度（１０、１００、１０００μｇ／ｍＬ）の発酵処理物を含む培養液Ａを調製した。調製した各試験液２００μＬと各ウェルの培養液Ａを置換した。置換後、カルチャースライドを１０秒間攪拌し、３７℃に設定した５％ＣＯ２インキュベーターに移し、１時間培養を行った。なお、陰性対照群として、発酵処理物を含まない培養液Ａを使用した。
２×１０６個のポリスチレンビーズを含む培養液Ａ５．０μＬを加えた後、３７℃、５％ＣＯ２インキュベーター内で２時間培養した。
A culture solution A containing the fermented product at each concentration (10, 100, 1000 μg / mL) was prepared. The prepared test solution (200 μL) and the culture solution A in each well were replaced. After the replacement, the culture slide was stirred for 10 seconds, transferred to a 5% CO 2 incubator set at 37 ° C., and cultured for 1 hour. In addition, the culture solution A which does not contain a fermentation processed material was used as a negative control group.
After adding 5.0 μL of a culture solution A containing 2 × 10 6 polystyrene beads, the cells were cultured for 2 hours in a 37 ° C., 5% CO 2 incubator.
カルチャースライドを振盪した後、液をアスピレータで除き、ウェルに３７℃に加温したＰＢＳ（−）を４００μL加えて、再度アスピレータで液を除く洗浄操作を計２回行った。４％パラホルムアルデヒド含有ＰＢＳ（−）溶液を２００μＬ加え、１時間室温で固定し、４℃で保存した。 After shaking the culture slide, the liquid was removed with an aspirator, 400 μL of PBS (−) heated to 37 ° C. was added to the well, and the washing operation for removing the liquid with the aspirator was performed twice in total. 200 μL of 4% paraformaldehyde-containing PBS (−) solution was added, fixed at room temperature for 1 hour, and stored at 4 ° C.
保存していたカルチャースライドのウェル中の液をアスピレータで除き、蒸留水を４００μＬ加え、再度アスピレータで液を除く洗浄操作を計３回行った後、カルチャースライドから仕切りを外し風乾させ、キシレン処理（キシレンを細胞にかけて、細胞に接着しているが、取り込まれていないビーズを除く操作）を行い、その後、風乾させた。ディフ・クイックで染色し、顕微鏡下（４００倍の倍率）で細胞に含まれるビーズを計測した。ビーズ数が細胞当たり１０個まではカウントした数値をそのまま記録し、細胞当たり１１個以上の場合は正確なカウントが困難な場合があるので、１０個以上と記載し、１０個として処理した。細胞数の合計が１００個になるまでカウントした。 Remove the liquid in the wells of the stored culture slide with an aspirator, add 400 μL of distilled water, and again perform the washing operation to remove the liquid with an aspirator three times. Remove the partition from the culture slide, air dry, and treat with xylene ( Xylene was applied to the cells to remove the beads that were adhered to the cells but not taken in, and then air-dried. The cells were stained with Diff-Quick and the beads contained in the cells were counted under a microscope (400 times magnification). When the number of beads was up to 10 per cell, the counted value was recorded as it was, and when it was 11 or more per cell, accurate counting might be difficult, so it was described as 10 or more and processed as 10. Counting was performed until the total number of cells reached 100.
カウントした値に基づいて、貪食率（一個以上のビーズを取り込んだ細胞数の全体の細胞数に対する割合）と貪食係数(細胞一個当たりの平均取り込み数)を算出した。 Based on the counted values, the phagocytosis rate (the ratio of the number of cells that took in one or more beads to the total number of cells) and the phagocytosis coefficient (the average number of cells taken up per cell) were calculated.
（３）結果
貪食率の比較では、陰性対照群に対して１０μｇ／ｍＬ発酵処理物添加群が１９．３％と約２．１倍、１００μｇ／ｍＬ発酵処理物添加群が１８．８％と約２．１倍、１０００μｇ／ｍＬ発酵処理物添加群が１９．０％と約２．１倍、高い値を示した（図１）。
貪食係数の比較においては、１０μｇ／ｍＬ発酵処理物添加群が０．４６（陰性対照群に対して約２．７倍）、１００μｇ／ｍＬ発酵処理物添加群が０．４３（陰性対照群に対して約２．５倍）、１０００μｇ／ｍＬ発酵処理物添加群が０．３３（陰性対照群に対して約２．０倍）と高い値を示した（図２）。なお、データを示していないが、本試験では、陰性対照群と比較して陽性対照群（１μｇ／ｍＬのＬＰＳを含む培養液Ａ）が有意に高い貪食率および貪食係数が認められたので、試験の成立条件を満たしていると判断した。
(3) Results In the comparison of the phagocytosis rate, the 10 μg / mL fermented product added group was 19.3% and about 2.1 times the negative control group, and the 100 μg / mL fermented product added group was 18.8%. About 2.1 times, the 1000 μg / mL fermented product addition group showed a high value, about 2.1 times, 19.0% (FIG. 1).
In comparison of the phagocytic coefficient, the 10 μg / mL fermented product added group was 0.46 (about 2.7 times the negative control group), and the 100 μg / mL fermented product added group was 0.43 (negative control group). In contrast, the 1000 μg / mL fermented product addition group showed a high value of 0.33 (about 2.0 times that of the negative control group) (FIG. 2). Although data is not shown, in this test, the phagocytosis rate and phagocytic coefficient were significantly higher in the positive control group (culture solution A containing 1 μg / mL LPS) than in the negative control group. It was judged that the test conditions were met.
〔５：免疫増強作用（２）〕
本発明の酵素処理物によるマクロファージ様細胞のＴＮＦ−α産生量増加効果を検討した。
[5: Immunity enhancing action (2)]
The effect of increasing the TNF-α production of macrophage-like cells by the enzyme-treated product of the present invention was examined.
（１）前培養
マウス由来マクロファージ様細胞株ＲＡＷ２６４．７細胞（ＥＣＡＣＣ ９１０６２０７２）は、４５００ｍｇ／Ｌグルコースおよび１０％ＦＢＳ含有ＤＭＥＭ培地（以下、培養液Ｂという。）にて継代培養したものを用いた。培養には１０ｃｍシャーレを用い、２または３日毎に１〜２×１０６ｃｅｌｌｓ／ｐｌｅｔｅで植え継いだ。３７℃に設定した５％ＣＯ２インキュベーター内で培養した。
(1) Preculture The mouse-derived macrophage-like cell line RAW264.7 cell (ECACC 91062072) was subcultured in DMEM medium (hereinafter referred to as culture medium B) containing 4500 mg / L glucose and 10% FBS. It was. A 10 cm petri dish was used for the culture, and the cells were planted at 1-2 × 10 6 cells / plete every 2 or 3 days. The cells were cultured in a 5% CO 2 incubator set at 37 ° C.
（２）試験操作
以下、全ての試験操作はクリーンベンチ内で行った。
１０ｃｍシャーレにて前培養したＲＡＷ２６４．７細胞をピペッティグと０．２５％トリプシンにより剥がし、培養液Ｂと合わせて１０ｍＬに調整して得られた細胞の懸濁液を５０ｍＬ遠沈管に移した。懸濁液から５０μＬを取り出し、同量のトリパンブルーと混ぜ合わせた後、細胞数をカウントした。遠沈管を室温で、１０００ｒｐｍ（１９０×ｇ）５分間遠心を行い、上清を捨て、細胞をペレットとして回収した。
(2) Test operation Hereinafter, all test operations were performed in a clean bench.
The RAW264.7 cells pre-cultured in a 10 cm petri dish were peeled off with pipetting and 0.25% trypsin, and the suspension of cells obtained by adjusting to 10 mL with the culture solution B was transferred to a 50 mL centrifuge tube. After removing 50 μL from the suspension and mixing with the same amount of trypan blue, the number of cells was counted. The centrifuge tube was centrifuged at room temperature at 1000 rpm (190 × g) for 5 minutes, the supernatant was discarded, and the cells were collected as a pellet.
測定した細胞数に基づいて培養液Ｂを加え、ピペッティングで均一に懸濁して１×１０５ｃｅｌｌ／ｍＬの懸濁液を調整した。得られた細胞懸濁液を１ｍＬずつ２４ｗｅｌｌ ｐｌａｔｅの各ウェルに加えた。３７℃の５％ＣＯ２インキュベーターに移して、細胞がウェルの底に接着して伸展するまで２４時間前培養を行った。 Culture medium B was added based on the measured number of cells, and the suspension was uniformly suspended by pipetting to prepare a suspension of 1 × 10 5 cells / mL. 1 mL of the obtained cell suspension was added to each well of a 24 well plate. The cells were transferred to a 37 ° C. 5% CO 2 incubator and pre-cultured for 24 hours until the cells adhered to the bottom of the well and spread.
各濃度（１、２．５、５ｍｇ／ｍＬ）の酵素処理物を含む培養液Ｂおよび各濃度（０．１、０．５、１、２．５、５ｍｇ／ｍＬ）の発酵処理物を含む培養液Ｂを調製した。調製した各試験液１ｍＬと各ウェルの培養液を置換した。置換後、３７℃に設定した５％ＣＯ２インキュベーターに移し、２４時間培養を行った。なお、陰性対照群として、発酵処理物を含まない培養液Ｂを使用した。 Includes culture solution B containing enzyme-treated product at each concentration (1, 2.5, 5 mg / mL) and fermentation-treated product at each concentration (0.1, 0.5, 1, 2.5, 5 mg / mL) Culture solution B was prepared. 1 mL of each prepared test solution and the culture solution in each well were replaced. After the replacement, it was transferred to a 5% CO 2 incubator set at 37 ° C. and cultured for 24 hours. In addition, the culture solution B which does not contain a fermentation processed material was used as a negative control group.
各試験液を各ウェルから１５０μＬ回収した。回収した各試験液それぞれに関して、専用のキット（Ｒ＆Ｄｓｙｓｔｅｍｓ Ｑｕａｎｔｉｋｉｎｅ Ｍｏｕｓｅ ＥＬＩＳＡｋｉｔ）を用いて評価を行った。 150 μL of each test solution was collected from each well. Each collected test solution was evaluated using a dedicated kit (R & D systems Quantikine Mouse ELISA kit).
（３）結果
酵素処理物はＲＡＷ２６４．７細胞のＴＮＦ−α産生量を増加させることが確認された（図３）。発酵処理物は濃度依存的（０．１ｍｇ／ｍＬで２４．９ｐｇ／ｍＬ、０．５ｍｇ／ｍＬで３５．１ｐｇ／ｍＬ、１ｍｇ／ｍＬで３６．４ｐｇ／ｍＬ、２．５ｍｇ／ｍＬで５６．９ｐｇ／ｍＬ、５ｍｇ／ｍＬで１１５．７ｐｇ／ｍＬ）にＲＡＷ２６４．７細胞のＴＮＦ−α産生量を増加させることが確認された（図４）。
また、発酵処理物と酵素処理物との同濃度における効果を比較すると、発酵処理物は酵素処理物よりもＲＡＷ２６４．７細胞のＴＮＦ−α産生量を増加させる効果が高いことが確認された（図３、４）。なお、データを示していないが、本試験では、陰性対照群と比較して陽性対照群（１μｇ／ｍＬのＬＰＳを含む培養液Ｂ）は高いＴＮＦ−α産生値を示したので、試験の成立条件を満たしていると判断した。
(3) Results It was confirmed that the enzyme-treated product increased the amount of TNF-α produced by RAW264.7 cells (FIG. 3). The fermented product was concentration-dependent (24.9 pg / mL at 0.1 mg / mL, 35.1 pg / mL at 0.5 mg / mL, 36.4 pg / mL at 1 mg / mL, 56. 9 pg / mL, 55.7 mg / mL (115.7 pg / mL) was confirmed to increase TNF-α production in RAW264.7 cells (FIG. 4).
Moreover, when the effect in the same density | concentration with a fermented processed material and an enzyme processed material was compared, it was confirmed that the fermented processed material has the effect of increasing the TNF- (alpha) production amount of RAW264.7 cell rather than an enzyme processed material ( 3 and 4). Although data is not shown, in this test, the positive control group (culture solution B containing 1 μg / mL LPS) showed a higher TNF-α production value than the negative control group. Judged that the conditions were met.
〔６：ＡＴＰ産生増強作用〕
本発明の酵素処理物による、マウス骨格筋由来細胞のＡＴＰ産生量増加効果を検討した。
[6: ATP production enhancing action]
The effect of increasing the amount of ATP produced in mouse skeletal muscle-derived cells by the enzyme-treated product of the present invention was examined.
（１）前培養
マウス骨格筋由来細胞株Ｃ２Ｃ１２細胞（ＥＣＡＣＣ ＲＣＢ０９８７）は、１０％ＦＢＳ含有ＤＭＥＭ培地（以下、培養液Ｃという。）にて継代培養したものを用いた。培養には７５ｃｍ２培養フラスコを用い、３または４日毎に２〜４×１０４ｃｅｌｌｓ／ｍＬで植え継いだ。３７℃に設定した５％ＣＯ２インキュベーター内で培養した。
(1) Preculture The mouse skeletal muscle cell line C2C12 cells (ECACC RCB0987) were subcultured in 10% FBS-containing DMEM medium (hereinafter referred to as culture medium C). A 75 cm 2 culture flask was used for the culture, and the cells were subcultured at 2 to 4 × 10 4 cells / mL every 3 or 4 days. The cells were cultured in a 5% CO 2 incubator set at 37 ° C.
（２）試験操作
以下、全ての試験操作はクリーンベンチ内で行った。
７５ｃｍ２培養フラスコにて前培養したＣ２Ｃ１２細胞に０．２５％トリプシンを含有するＰＢＳ（−)溶液を加え、ピペッティングにより剥がし、得られた細胞の懸濁液を５０ｍＬ遠沈管に移した。遠沈管を室温で、１０００ｒｐｍ、３分間遠心を行い、上清を捨て、培地を１０ｍＬ添加し懸濁した。細胞懸濁液から５０μＬを取り、同量のトリパンブルーと混ぜ合わせた後、細胞数をカウントした。遠沈管を室温で、１０００ｒｐｍ、３分間遠心を行い、上清を捨て、細胞をペレットとして回収した。
(2) Test operation Hereinafter, all test operations were performed in a clean bench.
A PBS (−) solution containing 0.25% trypsin was added to C2C12 cells pre-cultured in a 75 cm 2 culture flask, peeled off by pipetting, and the resulting cell suspension was transferred to a 50 mL centrifuge tube. The centrifuge tube was centrifuged at room temperature at 1000 rpm for 3 minutes, the supernatant was discarded, and 10 mL of the medium was added and suspended. After taking 50 μL from the cell suspension and mixing with the same amount of trypan blue, the number of cells was counted. The centrifuge tube was centrifuged at 1000 rpm for 3 minutes at room temperature, the supernatant was discarded, and the cells were collected as a pellet.
測定した細胞数に基づいて培養液Ｃを加え、ピペッティングで均一に懸濁して１×１０４ｃｅｌｌｓ／ｍＬの細胞懸濁液に調製した。ＡＴＰ産生量測定用には９６ウェルホワイトプレートに、タンパク質定量用には９６ウェルプレートに、細胞懸濁液をそれぞれ０．１ｍＬずつ播種した。３７℃、５％ＣＯ２インキュベーターに移して、細胞がウェルの底に接着して伸展するまで２４時間前培養を行った。 Culture medium C was added based on the measured number of cells, and the suspension was uniformly suspended by pipetting to prepare a cell suspension of 1 × 10 4 cells / mL. 0.1 mL each of the cell suspension was seeded in a 96-well white plate for measuring ATP production and in a 96-well plate for protein quantification. The cells were transferred to a 37 ° C., 5% CO 2 incubator and pre-cultured for 24 hours until the cells adhered to the bottom of the well and spread.
各濃度（０．１、０．５、１．０ｍｇ／ｍＬ）の発酵処理物を含む培養液Ｃを調製した。調製した各試験液０．１ｍＬと各ウェルの培養液を置換した。置換後、３７℃、５％ＣＯ２インキュベーターに移し、２４時間培養を行った。なお、陰性対照群として、発酵処理物を含まない培養液Ｃを使用した。 The culture solution C containing the fermented product at each concentration (0.1, 0.5, 1.0 mg / mL) was prepared. The prepared test solution (0.1 mL) and the culture solution in each well were replaced. After the replacement, the cells were transferred to a 37 ° C., 5% CO 2 incubator and cultured for 24 hours. In addition, the culture solution C which does not contain a fermentation processed material was used as a negative control group.
ＡＴＰ産生量測定用プレートの各ウェルに、細胞のＡＴＰ測定試薬（東洋ビーネット製）を０．１ｍＬずつ添加し、ルミノメーター（ＡＲＶＯ ＭＸ、ＰｅｒｋｉｎＥｌｍｅｒ製）を用いて発光量を測定した。タンパク質定量用プレートはＢＣＡ ｐｒｏｔｅｉｎ Ａｓｓａｙ ｋｉｔ（サーモサイエンティフィック製）を用いて、ウェル中の総タンパク質定量を行った。 0.1 mL of cell ATP measurement reagent (Toyo Benet) was added to each well of the ATP production amount measurement plate, and the amount of luminescence was measured using a luminometer (ARVO MX, manufactured by PerkinElmer). The protein quantification plate used was BCA protein Assay kit (manufactured by Thermo Scientific) to quantitate the total protein in the wells.
（３）結果
細胞あたりのＡＴＰ産生量の比較では、０．１および０．５ｍｇ／ｍＬ発酵処理物添加群が陰性対照群に対して約１．５倍であり、１．０ｍｇ／ｍＬ発酵処理物添加群が陰性対照群に対して約１．６倍であった（図５）。なお、データを示していないが、本試験では、陰性対照群と比較して陽性対照群（２ｎＭピルビン酸を含む培養液Ｃ）が有意に高いＡＴＰ産生値を示したので、試験の成立条件を満たしていると判断した。
(3) Results In the comparison of the amount of ATP produced per cell, the 0.1 and 0.5 mg / mL fermented product addition groups were about 1.5 times the negative control group, and the 1.0 mg / mL fermented treatment The product addition group was about 1.6 times the negative control group (FIG. 5). Although the data is not shown, in this test, the positive control group (culture medium C containing 2 nM pyruvic acid) showed significantly higher ATP production value than the negative control group. Judged that it met.
〔７：ミトコンドリア関連遺伝子（ＵＣＰ３遺伝子、ＰＧＣ１α遺伝子）の発現解析〕
本発明の酵素処理物刺激による、ミトコンドリア関連遺伝子（ＵＣＰ３遺伝子、ＰＧＣ１α遺伝子）の発現への影響を調べた。
[7: Expression analysis of mitochondria-related genes (UCP3 gene, PGC1α gene)]
The influence on the expression of mitochondria-related genes (UCP3 gene, PGC1α gene) by stimulation with the enzyme-treated product of the present invention was examined.
（１）前培養
Ｃ２Ｃ１２細胞は、培養液Ｃにて継代培養したものを用いた。培養には１０ｃｍ２細胞培養用シャーレを用い、３または４日毎に１〜２×１０５ｃｅｌｌｓ／ｍＬで植え継いだ。３７℃に設定した５％ＣＯ２インキュベーター内で培養した。
(1) Pre-culture C2C12 cells subcultured in culture medium C were used. A 10 cm 2 cell culture petri dish was used for the culture, and the cells were planted at 1-2 × 10 5 cells / mL every 3 or 4 days. The cells were cultured in a 5% CO 2 incubator set at 37 ° C.
（２）試験操作
以下の試験操作はクリーンベンチ内で行った。
１０ｃｍ２細胞培養用シャーレにて前培養したＣ２Ｃ１２細胞に０．２５%トリプシンを含有したＰＢＳ（−）溶液を加え反応後、ピペッティングにより細胞を回収した。得られた細胞の懸濁液を５０ｍＬ遠沈管に移し、室温で１０００ｒｐｍ（１９０×ｇ）、３分間遠心を行った。上清を捨て、細胞をペレットとして回収した。
細胞数を計測し、５×１０４ｃｅｌｌｓ／ｍＬの濃度に調整後、２４ウェルプレートの各ウェルに５００μＬずつ加えた。３７℃、５％ＣＯ２インキュベーターに移して、細胞がウェルの底に接着して伸展するまで２日間前培養を行った。分化誘導培地（２％ＦＢＳ含有ＤＭＥＭ培地）に交換し、さらに４日間培養した。
(2) Test operation The following test operation was performed in a clean bench.
A PBS (−) solution containing 0.25% trypsin was added to C2C12 cells pre-cultured in a 10 cm 2 cell culture dish, and then the cells were collected by pipetting. The obtained cell suspension was transferred to a 50 mL centrifuge tube and centrifuged at 1000 rpm (190 × g) for 3 minutes at room temperature. The supernatant was discarded and the cells were collected as a pellet.
The number of cells was counted, adjusted to a concentration of 5 × 10 4 cells / mL, and 500 μL was added to each well of a 24-well plate. The cells were transferred to a 37 ° C., 5% CO 2 incubator and precultured for 2 days until the cells adhered to the well bottom and spread. The medium was replaced with a differentiation induction medium (DMEM medium containing 2% FBS), and further cultured for 4 days.
各濃度（０．１、０．５、１．０ｍｇ／ｍＬ）の酵素処理物を含む培養液Ｃおよび各濃度（０．１、０．５、１．０ｍｇ／ｍＬ）の発酵処理物を含む培養液Ｃを調製した。調製した各試験液５００μＬと各ウェルの培養液Ｃを置換し、３７℃、５％ＣＯ２インキュベーターに移して、６時間培養した。培養液Ｃを除き、細胞を３００μＬのＩＳＯＧＥＮに（ニッポンジーン製）に溶解した。添付のプロトコルに準じて、ｔｏｔａｌ ＲＮＡを抽出した。なお、陰性対照群として、発酵処理物を含まない培養液Ｃを使用した。 Includes culture medium C containing enzyme-treated product at each concentration (0.1, 0.5, 1.0 mg / mL) and fermentation-treated product at each concentration (0.1, 0.5, 1.0 mg / mL) Culture solution C was prepared. The prepared 500 μL of each test solution and the culture solution C in each well were replaced, transferred to a 37 ° C., 5% CO 2 incubator, and cultured for 6 hours. The culture medium C was removed, and the cells were dissolved in 300 μL of ISOGEN (manufactured by Nippon Gene). Total RNA was extracted according to the attached protocol. In addition, the culture solution C which does not contain a fermentation processed material was used as a negative control group.
得られたｔｏｔａｌ ＲＮＡは分光光度計を用いて濃度を測定し、５００ｎｇの ｔｏｔａｌ ＲＮＡを鋳型とし、ＰｒｉｍｅＳｃｒｉｐｔ ＲＴ ｒｅａｇｅｎｔ Ｋｉｔ（タカラバイオ製）を用いてｃＤＮＡを合成した。得られたｃＤＮＡを鋳型とし、ＳＹＢＲ Ｐｒｅｍｉｘ ＥＸ Ｔａｑ に（タカラバイオ製）およびＬｉｇｈｔＣｙｃｌｅｒ ４８０（ロシュ・ダイアグノスティックス製）を用いてリアルタイムＰＣＲを行い、ＵＣＰ３、ＰＧＣ１α遺伝子の発現量を測定した。また、内部標準遺伝子としてはＧＡＰＤＨの発現量を定量した。定量に用いたプライマーはＢｉｏｓｃｉ． Ｒｅｐ．３２，４６５−４７８（２０１２）に記載された配列をもとに作製した（表１参照）。
ＬｉｇｈｔＣｙｃｌｅｒ ４８０専用ソフトウェア（ロシュ・ダイアグノスティックス製）を用いて解析した。
The concentration of the obtained total RNA was measured using a spectrophotometer, and 500 ng of total RNA was used as a template, and cDNA was synthesized using PrimeScript RT reagent Kit (manufactured by Takara Bio Inc.). Using the obtained cDNA as a template, real-time PCR was performed using SYBR Premix EX Taq (manufactured by Takara Bio) and LightCycler 480 (manufactured by Roche Diagnostics), and the expression levels of UCP3 and PGC1α genes were measured. Moreover, the expression level of GAPDH was quantified as an internal standard gene. Primers used for quantification were Biosci. Rep. 32, 465-478 (2012), based on the arrangement | sequence (refer Table 1).
Analysis was performed using LightCycler 480 dedicated software (Roche Diagnostics).
（３）結果
酵素処理物
ミトコンドリア関連遺伝子のうち、ＵＣＰ３遺伝子の発現量の比較では、０．１ｍｇ／ｍＬ酵素処理物添加群が陰性対照群に対して約１．１倍、０．５ｍｇ／ｍＬ酵素処理物添加群が陰性対照群に対して約１．３倍、１．０ｍｇ／ｍＬ酵素処理物添加群が陰性対照群に対して約１．５倍であり、増加傾向（Ｐ＝０．０６）が認められた（図６）。
(3) Results
Among the enzyme-treated mitochondrial-related genes, in the comparison of the expression level of the UCP3 gene, the 0.1 mg / mL enzyme-treated product addition group was about 1.1 times the negative control group, and the 0.5 mg / mL enzyme-treated product addition The group was about 1.3 times that of the negative control group, and the 1.0 mg / mL enzyme-treated product-added group was about 1.5 times that of the negative control group, showing an increasing trend (P = 0.06). (FIG. 6).
発酵処理物
ミトコンドリア関連遺伝子のうち、ＵＣＰ３遺伝子の発現量の比較では、０．１ｍｇ／ｍＬ発酵処理物添加群が陰性対照群に対して約１．４倍、０．５ｍｇ／ｍＬ添加群が陰性対照群に対して約１．５倍、１．０ｍｇ／ｍＬ添加群が陰性対照群に対して約１．５倍であった（図７）。
ＰＧＣ１α遺伝子の発現量の比較では、０．１ｍｇ／ｍＬ発酵処理物添加群が陰性対照群に対して約１．１倍、０．５ｍｇ／ｍＬ発酵処理物添加群が陰性対照群に対して約１．２倍、１．０ｍｇ／ｍＬ発酵処理物添加群が陰性対照群に対して約１．３倍であり、増加傾向が認められた（図８）。
Among the fermented processed mitochondrial-related genes, in the comparison of UCP3 gene expression levels, the 0.1 mg / mL fermented product added group was about 1.4 times the negative control group, and the 0.5 mg / mL added group was negative About 1.5 times the control group and 1.0 mg / mL addition group were about 1.5 times the negative control group (FIG. 7).
In the comparison of the expression level of the PGC1α gene, the 0.1 mg / mL fermented product added group was about 1.1 times the negative control group, and the 0.5 mg / mL fermented product added group was about the negative control group. The 1.2 times, 1.0 mg / mL fermented product addition group was about 1.3 times the negative control group, and an increasing tendency was observed (FIG. 8).
なお本発明は上述した各実施形態および実施例に限定されるものではなく、請求項に示した範囲で種々の変更が可能であり、異なる実施形態にそれぞれ開示された技術的手段を適宜組み合わせて得られる実施形態についても本発明の技術的範囲に含まれる。また、本明細書中に記載された学術文献および特許文献の全てが、本明細書中において参考として援用される。 The present invention is not limited to the above-described embodiments and examples, and various modifications are possible within the scope shown in the claims, and technical means disclosed in different embodiments are appropriately combined. The obtained embodiment is also included in the technical scope of the present invention. Moreover, all the academic literatures and patent literatures described in this specification are incorporated herein by reference.
本発明のバナナ酵素処理物は、免疫増強剤、ミトコンドリア活性化促進剤として有用である。本発明のバナナ酵素処理物は飲食品、サプリメント等の成分として有用である。 The banana enzyme-treated product of the present invention is useful as an immune enhancer and a mitochondrial activation promoter. The banana enzyme-treated product of the present invention is useful as a component for foods and drinks, supplements and the like.
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