JP6712819B2 - Bioactive substance containing composition derived from banana - Google Patents

Description

The present invention relates to a method for producing a banana-derived composition and a physiologically active substance containing the banana-derived composition.

Banana is a plant belonging to the family Musaceae that is widely cultivated all over the world. In recent years, it has been found that each part (flower, root) of banana exerts various physiological activities, and such physiological activities have been attracting attention. Examples of such physiological activity include a blood sugar level elevation suppressing effect. In recent years, the number of adult diabetic patients has reached hundreds of millions worldwide, and insulin administration, islet transplantation, and exercise are effective methods for suppressing the rise in blood glucose levels and improving diabetes. .. On the other hand, it has been reported that an extract obtained from a banana flower or a root contributes to the improvement of hyperglycemia (see, for example, Non-Patent Documents 1 and 2). Specifically, Non-Patent Document 1 reports that an extract obtained from banana root has a diabetes improving effect. In addition, Non-Patent Document 2 reports that an extract obtained from a banana flower has an effect of lowering blood glucose level.

Salau B.A et al. Asian J. Exp. Biol. Sci., vol 1 (1): 30-35, 2010. L. Pari and J. Umamaheswari Phytother. Res 14: 136-138, 2000.

However, banana roots and flowers are difficult to obtain, and there has been a demand for cheaper and more stable supply. In addition, the mechanism by which the extracts of banana root and flowers give physiological activities such as the inhibitory effect on blood sugar level elevation was also unknown.

The present invention has been made in view of such circumstances, and an object of the present invention is to provide a method for producing a banana-derived composition from which the composition exhibiting excellent physiological activity can be obtained from the pulp part of the banana. And
Another object of the present invention is to provide a physiologically active substance containing a banana-derived composition obtained from the pulp portion of a banana, which exhibits excellent physiological activity.

The present inventor has paid attention to the flesh portion of a banana, which has already established a distribution route in the market and can secure a considerable amount at a low price, and has conducted earnest studies to achieve the above object. Then, the present inventor has found that a composition obtained by enzymatically decomposing the protein in the pulp portion of banana exhibits excellent physiological activity, and completed the present invention.

That is, the present invention has an object to advantageously solve the above problems, and a method for producing a banana-derived composition of the present invention comprises a step of subdividing a flesh part of a banana to obtain a subdivided flesh (1 ), and a step (2) of adding a proteolytic enzyme to the pulp fragmentation product to decompose the protein in the pulp fragmentation product by an enzymatic reaction to obtain an enzyme reaction product, and the protein degradation in the enzyme reaction product. A step (3) of deactivating the enzyme to obtain the composition A. When the banana-derived composition is produced by the production method including such steps, the obtained banana-derived composition can have excellent effects of suppressing an increase in blood sugar level and promoting collagen production.

Here, in the method for producing a banana-derived composition of the present invention, it is preferable that the pulp portion of the banana is a pulp portion of an unripe banana. This is because if the pulp portion of an immature banana is used as the pulp portion of the banana, the resulting banana-derived composition can have excellent physiological activities such as a blood sugar level increase suppressing action and a collagen production promoting action. In addition, the production efficiency can be significantly improved especially when an alcohol having 1 to 4 carbon atoms is used for deactivating the protease.

Then, in the method for producing a banana-derived composition of the present invention, in the step (3), it is preferable to deactivate the proteolytic enzyme by heating. This is because the deactivation of the proteolytic enzyme by heating can be performed with equipment that is easy and simple to operate, and the production efficiency can be improved.

Further, in the method for producing a banana-derived composition of the present invention, in the step (3), the proteolytic enzyme is deactivated by an alcohol having 1 to 4 carbon atoms, and the step (3) is performed by the enzyme. A step [I] in which the alcohol having 1 to 4 carbon atoms is added to the reaction product, and then the obtained alcohol additive is subjected to solid-liquid separation to take out a liquid phase, and the carbon number in the liquid phase is 1 It is preferable to include the step [II] of removing the alcohol of ~4. This is because if the proteolytic enzyme is inactivated with an alcohol having 1 to 4 carbon atoms, the active ingredient can be extracted with the alcohol and the yield of the obtained active ingredient can be improved. ..

In addition, in the method for producing a banana-derived composition of the present invention, it is preferable that the alcohol having 1 to 4 carbon atoms is at least one of methanol and ethanol. Methanol and ethanol having a lower boiling point than other alcohols can be easily removed by evaporation, and if methanol and ethanol having such properties are used, the heat history of the resulting banana-derived composition is reduced and physiological activity is reduced. This is because it can be excellent. In addition, methanol and ethanol are excellent in the extraction efficiency of the amino acid and tryptophan derivative in the pulp of banana.

Here, in the method for producing a banana-derived composition of the present invention, the step [II] is preferably a step of removing the alcohol having 1 to 4 carbon atoms in the liquid phase by freeze-drying. This is because if lyophilization is adopted as a method for removing alcohol, the heat history of the obtained banana-derived composition can be reduced and the physiological activity can be made excellent.

Then, the method for producing a banana-derived composition of the present invention preferably further comprises a step (4) of desaccharifying and desalting the composition A to obtain a composition B. If this step is provided, in addition to making the obtained banana-derived composition excellent in blood sugar level increase suppressing action and collagen production promoting action, it makes the obtained banana-derived composition excellent in antioxidant action. Because you can.

The banana-derived composition obtained by the method for producing a banana-derived composition of the present invention has physiological activity.

Further, the present invention is intended to advantageously solve the above problems, the physiologically active substance of the present invention is characterized in that it contains a banana-derived composition obtained from the flesh of banana as an active ingredient. And Such a physiologically active substance has excellent physiological activity.

Here, as for the physiologically active substance of the present invention, the banana-derived composition is preferably obtained from the pulp portion of an unripe banana. This is because if the banana-derived composition is obtained from the pulp portion of an unripe banana, the physiological activity of the physiologically active substance of the present invention can be made excellent.

Furthermore, the physiologically active substance of the present invention, the composition derived from banana, by adding a proteolytic enzyme to the pulp portion of the banana, to decompose the protein in the pulp portion of the banana by an enzymatic reaction, then the proteolysis It is preferably obtained by heating or deactivating the enzyme with alcohol. This is because if the banana-derived composition is produced through an enzymatic reaction and inactivation by heating or alcohol, the physiologically active substance of the present invention can have excellent physiological activity.

In addition, in the physiologically active substance of the present invention, the total of the amino acid content and the tryptophan derivative content in the banana-derived composition is preferably 7% by mass or more. This is because if the total of the amino acid content and the tryptophan derivative content in the banana-derived composition is 7% by mass or more, the physiological activity of the physiologically active substance of the present invention can be made excellent.

In the physiologically active substance of the present invention, it is preferable that the amino acid contains phenylalanine and tryptophan, and the tryptophan derivative contains serotonin. This is because when the physiologically active substance contains phenylalanine, tryptophan, and serotonin, the physiologically active substance of the present invention can have excellent physiological activity.

In addition, the physiologically active substance of the present invention exhibits at least one of a blood glucose level increase suppressing action and a collagen production promoting action.

ADVANTAGE OF THE INVENTION According to this invention, the manufacturing method of the composition derived from a banana which can obtain the composition which shows the outstanding physiological activity from the pulp part of a banana can be provided.
Further, according to the present invention, it is possible to provide a physiologically active substance containing a banana-derived composition obtained from the pulp portion of a banana, which exhibits excellent physiological activity.

It is a figure which shows the measurement result (rat) of a blood sugar level increase suppression effect. It is a figure which shows the measurement result (human) of a blood glucose level increase inhibitory effect. It is another figure which shows the measurement result (human) of a blood sugar level increase suppression effect. It is a figure which shows the measurement result of a collagen production promotion effect.

Hereinafter, embodiments of the present invention will be described in detail.
The method for producing a banana-derived composition of the present invention is a method for producing a composition having excellent physiological activity using the pulp portion of banana as a raw material. Further, the physiologically active substance of the present invention contains a banana-derived composition obtained from the flesh of banana as an active ingredient. The banana-derived composition in the physiologically active substance of the present invention can be produced using, for example, the method for producing a banana-derived composition of the present invention.

<Method for producing composition derived from banana>
The method for producing a banana-derived composition of the present invention comprises a step (1) of subdividing a flesh part of a banana to obtain a subdivided flesh, and a proteolytic enzyme is added to the subdivided flesh to enzymatically react the flesh into pieces. Characterized in that it comprises a step (2) of degrading a protein in the compound to obtain an enzymatic reaction product, and a step (3) of deactivating the proteolytic enzyme in the enzymatic reaction product to obtain a composition A. And
In addition to the steps (1) to (3), the method for producing a banana-derived composition of the present invention further comprises a step (4) of desaccharifying and desalting the composition A to obtain a composition B. May be.
The banana-derived composition obtained by such a production method exhibits excellent physiological activity. Although the mechanism by which the banana-derived composition obtained by the method for producing a banana-derived composition of the present invention has excellent physiological activity is not clear, amino acids (particularly phenylalanine and tryptophan) and tryptophan derivatives (particularly phenylalanine and tryptophan) mainly obtained by enzymatic reaction are not known. It is speculated that serotonin) contributes to physiological activities (suppressing effect on blood sugar level elevation, promoting collagen production, and antioxidative effect). Then, according to the production method of the present invention, such amino acids and tryptophan derivatives can be efficiently collected, and a banana-derived composition containing them at a high concentration can be produced.
The amino acids that can be contained in the resulting banana-derived composition include arginine, lysine, histidine, phenylalanine, tyrosine, leucine, isoleucine, methionine, valine, alanine, glycine, proline, glutamic acid, serine, threonine, aspartic acid, tryptophan. Are listed.
The tryptophan derivative refers to a compound (or substance) other than an amino acid that can be derived from tryptophan, preferably a compound (or substance) other than an amino acid that can be derived from tryptophan in a biological reaction of the human body, and examples thereof include serotonin and melatonin.

In the present specification, "step (1) of subdividing the flesh portion of banana to obtain a flesh segmentation product" is referred to as "flesh flesh segmentation step (1)", and "proteolytic enzyme is added to the flesh segmentation product". The step (2) of adding and decomposing the protein in the pulp fragment by enzymatic reaction to obtain the enzyme reaction product is referred to as "enzyme reaction process (2)" and "deactivating the proteolytic enzyme in the enzyme reaction product". , Step (3) of obtaining composition A is referred to as "enzyme deactivation step (3)" and "step (4) of decomposing and desalting composition A to obtain composition B" It is abbreviated as "salt step (4)".

[Pulp segmentation step (1)]
(Subdivision)
In the method for producing a banana-derived composition of the present invention, first, the pulp portion of the banana is subdivided to obtain a subdivided pulp. By subdividing the flesh portion of the banana into a subdivided flesh product, the proteolytic enzyme can be efficiently reacted with the protein in the enzyme reaction step (2) described below. In the present invention, "subdivision" means that after removing the pericarp, the pulp portion, which is in a state without a missing portion, is crushed, cut, or the like to be divided into fragments having a smaller mass than that state, or It means to make a paste by crushing, crushing, cutting and the like. The shape of the pulp fragment obtained by subdivision is not particularly limited and may be granular or paste, but from the viewpoint of sufficiently ensuring the efficiency of the enzymatic reaction, the paste is particularly preferable. preferable.
When the obtained pulp fragmentation is a granular fragment, the average mass of the fragment is preferably 20 g or less, more preferably 15 g or less, and further preferably 10 g or less. When the average mass of the fragments is 20 g or less, the efficiency of the enzymatic reaction can be sufficiently ensured. The average mass of the fragments is not particularly limited, but is usually 0.1 g or more. The “average mass of fragments” can be calculated as the average value of the masses of 10 fragments arbitrarily selected when the number of fragments after fragmentation exceeds 10, and the number of fragments after fragmentation is When it is 10 or less, it can be determined as the average value of the masses of all the fragments.
The method for subdividing the pulp portion of the banana is not particularly limited, and examples thereof include a method using a cutter mill crusher and a hammer mill crusher.

(Banana flesh)
In the present invention, the flesh portion of a banana refers to a portion obtained by removing the skin of a banana fruit (when the banana is an edible banana, it corresponds to a so-called edible portion). The varieties of bananas used are not particularly limited, and examples include varieties such as giant cavendish and lacatan. These may be used alone or in combination of two or more. Among these, Giant Cavendish is preferable. The flesh portion of the banana can be obtained by removing the skin of the banana fruit using a known method.

In the present invention, the pulp portion of banana is preferably the pulp portion of unripe banana. Here, in the present invention, the term "immature banana" means that the fruit juice obtained by squeezing and filtering a paste obtained by crushing and cutting the pulp portion has a Brix degree of 24° when measured by a sugar refractometer. It is less than Bx. By using the pulp portion of the immature banana having a relatively low sugar content as the pulp portion of the banana, the sugar content of the obtained banana-derived composition (primary refined product) can be reduced, and as a result, The content (content ratio) of amino acids and tryptophan derivatives (particularly phenylalanine, tryptophan, and serotonin) in the composition derived from banana increases. That is, when the pulp portion of the unripe banana is used, the total amino acid content and tryptophan derivative content in the banana-derived composition can be increased as compared with the case where the pulp portion of the ripe banana is used. It is possible to improve physiological activity (suppressive action of blood sugar level increase, action of promoting collagen production). Further, the pulp portion of the immature banana is less sticky, and by using the pulp portion of the immature banana as the pulp portion of the banana, it becomes possible to improve the filtration efficiency in the manufacturing process (for example, the solid-liquid separation step described later ( It becomes easy to take out the liquid phase in I), and the production efficiency can be significantly improved.
And from the viewpoints of greatly improving the production efficiency, facilitating desugarization, and improving the physiological activity as described above, the brix degree of the flesh portion of the banana is preferably 10°Bx or less, more preferably 8°. Bx or less, even more preferably 5° Bx or less, particularly preferably 2° Bx or less. The lower limit of the Brix degree of the pulp portion of the banana is not particularly limited, but the Brix degree is usually 0.1°Bx or more.

[Enzyme reaction step (2)]
Next, a proteolytic enzyme is added to the obtained pulp fragmentation product to decompose the protein in the pulp fragmentation product by an enzymatic reaction to obtain an enzyme reaction product. By this enzymatic reaction, the amounts of amino acids and tryptophan derivatives (particularly phenylalanine, tryptophan, and serotonin) in the resulting banana-derived composition can be significantly increased, and physiological activity can be secured.

(Proteolytic enzyme)
The proteolytic enzyme used in the enzymatic reaction step (2) is not particularly limited as long as it can decompose the protein contained in the pulp portion of banana, and either endo type or exo type may be used. Examples of proteolytic enzyme agents containing endo-type proteolytic enzymes include protin SD-AY10 and protin SD-NY10 (both manufactured by Amano Enzyme Inc.). Examples of proteolytic enzyme agents containing exo-type proteolytic enzymes include Proteax (registered trademark), Protease M "Amano" SD, and Protease P "Amano" SD (all manufactured by Amano Enzyme Inc.).
The proteolytic enzyme to be used is preferably an exo-type proteolytic enzyme, and more preferably protease M "Amano" SD, from the viewpoint of being able to more efficiently decompose the protein in the pulp-digested product to generate an amino acid.

(Enzyme reaction)
In the enzymatic reaction step (2), the amount of the proteolytic enzyme added is preferably 0.5 part by mass or more, more preferably 1 part by mass or more, and particularly preferably 3 parts by mass or more, per 100 parts by mass of the pulp fragmentation product. , Preferably 20 parts by mass or less, more preferably 15 parts by mass or less, and particularly 10 parts by mass or less. When the amount of the proteolytic enzyme added is 0.5 parts by mass or more per 100 parts by mass of the pulp fragment, the enzymatic reaction can be favorably advanced, and when it is 20 parts by mass or less, the active ingredient that can be obtained It is superior in terms of both quantity and cost.

In addition, it is preferable to add water to the pulp fragmentation product prior to the addition of the proteolytic enzyme. The amount of water added to the pulp fragment is preferably 50 parts by mass or more, more preferably 100 parts by mass or more, particularly preferably 150 parts by mass or more, and preferably 500 parts by mass or less, per 100 parts by mass of the pulp fragmented product. , More preferably 300 parts by mass or less, particularly preferably 250 parts by mass or less. The amount of water added to the pulp fragment is 50 parts by mass or more per 100 parts by mass of the pulp fragment, so that the proteolytic enzyme can be uniformly diffused in the pulp fragment and the enzymatic reaction can be favorably progressed. You can On the other hand, when the amount of water added to the pulp fragment is 500 parts by mass or less per 100 parts by mass of the pulp fragment, the concentration of the proteolytic enzyme in the mixture becomes appropriate, and the enzymatic reaction can proceed satisfactorily. ..

The pH of the pulp fragmentation product (or the mixture of pulp fragmentation product and water) before the addition of the protease is preferably 4.5 to 6.0. When the pH of the pulp fragmentation product (or the mixture of the pulp fragmentation product and water) is within the above range, the enzymatic reaction can proceed well.

The reaction temperature of the enzyme reaction is preferably 40° C. or higher, more preferably 45° C. or higher, and preferably 65° C. or lower. When the reaction temperature of the enzymatic reaction is within the above range, the enzymatic reaction can proceed well.

The reaction time of the enzyme reaction is preferably 1 hour or longer, more preferably 3 hours or longer, particularly preferably 4 hours or longer, and preferably 20 hours or shorter. By setting the reaction time of the enzymatic reaction to 20 hours or less, the decomposition of serotonin in the obtained banana-derived composition can be sufficiently suppressed.

Then, the enzyme reaction step (2) comprises a first enzyme reaction step having a reaction temperature of 45° C. or higher and 55° C. or lower and a reaction time of 3 to 5 hours, and a reaction temperature of more than 55° C. and 65° C. or lower after the first enzyme reaction step. It is preferable to include the second enzymatic reaction step for 1 to 17 hours. As described above, the enzyme reaction step includes the first enzyme reaction step and the second enzyme reaction step, so that the enzyme reaction can proceed well.

[Enzyme deactivation step (3)]
Then, the proteolytic enzyme in the enzymatic reaction product obtained after the above-mentioned enzymatic reaction is deactivated. The method of deactivating the proteolytic enzyme in the enzyme reaction product is not particularly limited, and examples thereof include a method of deactivating by heating and a method of deactivating with an alcohol having 1 to 4 carbon atoms. These deactivating methods may be used in an appropriate combination.

(Deactivation due to heating)
First, a method for inactivating the protease to obtain the composition A by heating will be described in detail. Deactivation by heating can be performed with equipment that is easy and simple to operate, and the composition A is excellent in production efficiency.
Specifically, the enzyme reaction product obtained after the above-mentioned enzyme reaction is filtered, if necessary, and then heated. The temperature at the time of heating (enzyme reaction product temperature) is not particularly limited as long as it is a temperature at which the proteolytic enzyme can be deactivated, but is preferably 85°C or higher and 95°C or lower. When the temperature during heating is 85° C. or higher, deactivation can be sufficiently advanced, and when the temperature is 95° C. or lower, adverse effects due to thermal decomposition of the active ingredient can be suppressed.
The heating time is not particularly limited, but is preferably 10 minutes or more and 60 minutes or less. When the heating time is within the above range, deactivation can be completed sufficiently.

After deactivation by heating, water is removed if necessary to obtain a composition A (primary purified product). Here, when water is removed until the composition A becomes a dry solid (for example, the solid content concentration is 90% by mass or more), the composition A can be a banana-derived composition having the effect of the present invention. .. This composition A has an excellent effect of suppressing an increase in blood glucose level and an effect of promoting collagen production. Further, when water is removed until the composition A is in the state of a concentrated liquid (for example, the solid content concentration is 10% by mass or more and less than 90% by mass), the composition A has a desalting and desalting treatment step (4) described below. ) Can also be used as is.
The method for removing water is not particularly limited, and examples thereof include concentration, freeze drying, and spray drying. Among these, when the composition A is a dried product, freeze-drying is preferable from the viewpoint of reducing the thermal history of the obtained banana-derived composition and making the physiological activity excellent. Further, as the method for removing these waters, only one method may be used, or a plurality of methods may be combined. For example, it may be freeze-dried after being inactivated by heating and then concentrated, or may be inactivated by heating and spray-dried. These drying conditions can be adjusted appropriately. Then, an excipient may be added prior to this drying. The amount of the excipient added can be adjusted appropriately.

(Deactivation due to alcohol)
Next, a method for inactivating the proteolytic enzyme with an alcohol having 1 to 4 carbon atoms to obtain the composition A will be described in detail. The inactivation by the alcohol is excellent in the yield of the active ingredient in the obtained composition A. When deactivating with an alcohol having 1 to 4 carbon atoms, in the step (3), an alcohol having 1 to 4 carbon atoms is added to the enzymatic reaction product obtained after the enzymatic reaction, and then the obtained alcohol is obtained. A step [I] (hereinafter, appropriately abbreviated as "solid-liquid separation step [I]") in which the additive is solid-liquid separated and a liquid phase is taken out, and an alcohol having 1 to 4 carbon atoms in the liquid phase is removed. It is preferable to include a step [II] (hereinafter, appropriately abbreviated as “solvent removal step [II]”).

-Solid-liquid separation step [I]-
First, the proteolytic enzyme is deactivated by adding an alcohol having 1 to 4 carbon atoms to the enzyme reaction product. Further, the active ingredient can be extracted into the liquid phase containing the alcohol. In addition, by adding an alcohol having 1 to 4 carbon atoms to the enzyme reaction product, solid-liquid separation, particularly filtration described below can be easily performed.
In order to sufficiently extract the active ingredient in the enzymatic reaction product obtained by the enzymatic reaction, after addition of an alcohol having 1 to 4 carbon atoms, the mixture is allowed to stand at a temperature of 10 to 30° C. for 2 to 20 hours, and then a solid-liquid reaction is performed. It is preferable to separate them.

The alcohol having 1 to 4 carbon atoms used in the solid-liquid separation step [I] is an alcohol having 1 to 4 carbon atoms in its molecular structure, such as methanol, ethanol, 1-propanol, 2-propanol, 1- Butanol and the like can be mentioned. These may be used alone or in combination of two or more. Among these, at least one of methanol and ethanol is preferable. Methanol or ethanol having a lower boiling point than other alcohols can be easily removed by evaporation. Therefore, the heat history of the obtained banana-derived composition can be reduced, and the physiological activity such as the blood sugar level elevation suppressing effect can be made excellent. In addition, methanol and ethanol are superior to other alcohols in extraction efficiency of amino acid and tryptophan derivative in the pulp of banana. From the viewpoint of using the resulting banana-derived composition for food, it is more preferable to use ethanol.

In the solid-liquid separation step [I], the addition amount of the alcohol having 1 to 4 carbon atoms is not particularly limited, but for example, 100 parts by mass or more, and more preferably 200 parts by mass or more per 100 parts by mass of the pulp fragmentation product. It is particularly preferably 250 parts by mass or more, preferably 1500 parts by mass or less, more preferably 1200 parts by mass or less, even more preferably 400 parts by mass or less, and particularly preferably 350 parts by mass or less. When the amount of alcohol added is 100 parts by mass or more per 100 parts by mass of the pulp fragment, the solid-liquid separation efficiency of the alcohol additive is improved, and when it is 1500 parts by mass or less, the subsequent solvent removal step [II ], the removal of alcohol can be facilitated.

As a method for solid-liquid separation (separation into solid phase and liquid phase) of an alcohol additive obtained by adding an alcohol having 1 to 4 carbon atoms to an enzyme reaction product, the alcohol additive is solid phase and liquid phase. The method is not particularly limited as long as it can be separated into the liquid phase and the liquid phase can be taken out, and examples thereof include methods such as filtration and centrifugation. Of these, filtration is preferable from the viewpoint that solid-liquid separation can be reliably performed. Examples of the method for filtering the alcohol additive include natural filtration, suction filtration, pressure filtration and the like.

-Solvent removal step [II]-
Part or all of the alcohol having 1 to 4 carbon atoms in the liquid phase taken out in the solid-liquid separation step [I] is removed to obtain the composition A (primary purified product). When water is contained in the liquid phase, the water in the liquid phase may be partially or wholly removed together. Here, when the solvent (alcohol having 1 to 4 carbon atoms and arbitrary water) is removed until the composition A becomes a dry solid (for example, the solid content concentration is 90% by mass or more), the composition A is A banana-derived composition that exhibits the effects of the invention can be obtained. This composition A has an excellent effect of suppressing an increase in blood glucose level and an effect of promoting collagen production. Further, when the solvent is removed until the composition A is in the state of a concentrated liquid (for example, the solid content concentration is 10% by mass or more and less than 90% by mass), the composition A has a desalting and desalting treatment step (4) described below. ) Can also be used as is.
The method for removing the alcohol having 1 to 4 carbon atoms (and any water) in the liquid phase is not particularly limited, and examples thereof include concentration, freeze drying, and spray drying. Among these, when the composition A is a dried product, freeze-drying is preferable from the viewpoint of reducing the thermal history of the obtained banana-derived composition and making the physiological activity excellent. Further, as for the method of removing the alcohol having 1 to 4 carbon atoms (and any water), only one method may be used, or a plurality of methods may be combined. For example, the obtained liquid phase may be concentrated and then freeze-dried, or the obtained liquid phase may be concentrated and then spray-dried. Then, an excipient may be added prior to this drying. The amount of the excipient added can be adjusted appropriately.

The conditions for freeze-drying are not particularly limited, but for example, the obtained liquid phase (or the liquid phase after concentration, etc.) is first reserved for about 1 to 3 hours until the product temperature reaches -30°C to -25°C. After freezing, the product was controlled at a product temperature of -30 to -25°C under reduced pressure to a degree of vacuum of 40 kPa or less, then the shelf temperature was gradually raised to about 40°C, and then the obtained liquid phase (or after concentration) It is preferable that the drying is completed after the temperature of the liquid phase (1) and the like) have reached the same level as the shelf temperature for 3 hours or more.

[Desaturation and desalting step (4)]
Next, the method for producing a banana-derived composition of the present invention may include a step of desugarizing and desalting the composition A obtained above to obtain a composition B. By de-saccharifying and desalting the composition A, a sugar content in the composition A, a salt (inorganic salt, organic salt) and a part or all of free metal ions can be removed, and the resulting composition B ( The secondary purified product) exhibits an excellent anti-oxidation effect in addition to an excellent blood sugar level increase suppressing effect and collagen production promoting effect.

In the present invention, desaccharification and desalting may be carried out by independent and independent operations, or may be performed simultaneously by one operation.
As a method for desalting and desalting the composition A (a method for simultaneously performing desiccation and desalting in one operation), for example, the components contained in the composition A are adsorbed on an adsorption resin packed in a column, Examples include a method in which the adsorption resin is washed with water and then the components adsorbed to the adsorption resin are eluted from the adsorption resin with an eluent and taken out (hereinafter, appropriately abbreviated as “adsorption resin method”).
From the viewpoint of simplification of the process, it is preferable to carry out desaccharification and desalting simultaneously in one operation, and the above adsorption resin method is more preferable.
Hereinafter, the case where the above adsorption resin method is adopted in the desugarization and desalting step (4) will be described as an example.

In the adsorption resin method, a step of adding water to the composition A (water addition step), a composition A (or a composition in which water is added to the composition A (water additive)) is used as an adsorption resin method. Of adsorbing components (amino acids, etc.) contained in the composition A onto the adsorption resin (adsorption step), and passing water through the adsorption resin-filled column for purification. (Washing step), a step of passing the eluent through the column filled with the adsorption resin to elute the components adsorbed on the adsorption resin (elution step), and the elution step It is preferable to carry out the step of removing the eluent from the fraction (eluent removing step). Hereinafter, these steps will be specifically described.

(Water addition process)
Water is added to the composition A as needed. Specifically, when the composition A is a dry solid, it is necessary to add water to form an aqueous solution, and when the composition A is a concentrated solution containing a solvent, the concentrated solution is used as it is. However, water may be further added to adjust the solid content concentration. The solid content concentration of the composition A (or its water additive) before passing through the column filled with the adsorption resin is not particularly limited, but is preferably 3 to 30% by mass.

(Adsorption process)
The composition A (or its water additive) is passed through a column packed with an adsorption resin to adsorb components (amino acids, etc.) contained in the composition A to the adsorption resin.
Examples of the adsorbent resin include a synthetic adsorbent that adsorbs an organic substance from a solution by a physical interaction between pore surfaces in the resin and a non-adsorbent substance, and a cation exchange resin having a sulfonic acid group. Among these, a cation exchange resin is preferable, and a strongly acidic cation exchange resin is more preferable, because amino acids such as phenylalanine and tryptophan and tryptophan derivatives such as serotonin can be efficiently adsorbed. Examples of the strongly acidic cation exchange resin include Diaion (registered trademark) SK1B (manufactured by Mitsubishi Chemical Corporation) and Amberlite (registered trademark) IR120B (manufactured by Dow Chemical Company).

The flow rate of the composition A (or its water additive) that is passed through the column filled with the adsorption resin depends on the properties of the composition A (or its water additive) to be passed and the adsorption resin used. It may be appropriately determined, and although not particularly limited, the liquid is passed at a flow rate in the range of 0.5 to 5 SV, for example. The SV represents the amount of the solution passed through the column per unit time with respect to the adsorbent resin (volume basis at 25° C.), and the flow rate when the same volume of solution as the adsorbent resin is passed in 1 hour is 1 SV. And
The amount of the composition A passed through the column filled with the adsorption resin is not particularly limited, but is, for example, 1 to 30 RV. The RV represents the amount of the solution passed through the column with respect to the adsorption resin (volume basis at 25° C.), and the passing amount when the same volume of solution as the adsorption resin is passed is 1 RV.

(Washing process)
Next, water is passed through the column filled with the adsorption resin. By passing water through the column after the adsorption step, it is possible to remove non-adsorbed components and weakly adsorbed components (sugars, organic salts, inorganic salts, free metal ions, etc.), that is, desaccharification and desalting. It will be possible.
The flow rate of water to be passed through the column after the adsorption step may be appropriately determined according to the adsorption resin used, and is not particularly limited, but the flow rate is, for example, in the range of 0.5 to 5 SV. By allowing water to pass through at such a flow rate, it is possible to effectively perform desugaring and desalting.
The amount of water passing through the column after the adsorption step may be appropriately determined according to the adsorption resin used and is not particularly limited, but is, for example, 5 to 15 RV. By allowing such an amount of water to pass therethrough, desaturation and desalting can be sufficiently performed.

(Elution process)
Next, the eluent is passed through a column filled with the adsorption resin to elute the components adsorbed on the adsorption resin, and an elution fraction due to the dissolved and separated liquid is obtained. The eluent (also referred to as an eluent) is a liquid for eluting the components adsorbed on the adsorption resin from the adsorption resin. The eluent used is not particularly limited as long as it has the above function. For example, when a synthetic adsorbent such as SepaBeads (registered trademark) SP207 is used as the adsorption resin, the eluent may be a mixed solvent of water and alcohol (methanol and/or ethanol). The concentration of methanol and/or ethanol in the mixed solvent is not particularly limited, but is preferably 3 to 10% by mass. When the concentration of methanol and/or ethanol in the mixed solvent is within the above range, the components adsorbed on the adsorption resin can be efficiently eluted. Then, for example, when a strongly acidic cation exchange resin such as Diaion (registered trademark) SK1B is used as the adsorption resin, ammonia water can be used as the eluent. Although the ammonia concentration in the ammoni water is not particularly limited, it is preferably 0.1 to 5 mol/L. When the ammonia concentration in the ammonia water is within the above range, the components adsorbed on the adsorbent resin can be efficiently eluted.

The flow rate of the eluent that passes through the column in the elution step may be appropriately determined according to the adsorbent resin used, and is not particularly limited, but the eluent is passed at a flow rate in the range of 0.5 to 5 SV, for example. By passing water at such a flow rate, the components adsorbed on the adsorption resin can be efficiently eluted.
The amount of the eluent passed through the column in the elution step may be appropriately determined according to the adsorbent resin used and is not particularly limited, but is, for example, 5 to 60 RV. By passing such an amount of the eluent, the components adsorbed on the adsorbent resin can be more surely eluted.

(Eluent removal process)
The eluent is removed from the eluate fraction obtained in the elution step to obtain the composition B (dry purified product secondary purified product). The method of removing the eluent to obtain the dried composition B is not particularly limited, and examples thereof include the method described above in the section “solvent removal step [II]”. Among these, freeze-drying is preferable from the viewpoint of reducing the heat history of the obtained banana-derived composition and making the physiological activity excellent. As for the method of removing these eluents, only one method may be used as in the “solvent removal step [II]”, but a plurality of methods may be combined. For example, the obtained elution fraction may be concentrated and then freeze-dried, or the obtained elution fraction may be concentrated and then spray-dried.
The conditions for freeze-drying are not particularly limited, and the conditions described above in the section “solvent removal step [II]” are suitable.
The composition B thus obtained can be used as a banana-derived composition, and the composition B has an excellent blood glucose level suppressing action, a collagen production promoting action, and an antioxidant action.

<Bioactive substance>
The physiologically active substance of the present invention is characterized by containing a banana-derived composition obtained from the pulp part of banana as an active ingredient. This physiologically active substance exhibits excellent physiological activities such as an action of suppressing an increase in blood glucose level and an action of promoting collagen production.

[Banana-derived composition]
The banana-derived composition containing the physiologically active substance of the present invention as an active ingredient is not particularly limited as long as it is obtained from the banana pulp portion. Then, this banana-derived composition is preferably a composition obtained from the pulp portion of an unripe banana for the same reason as described above in “Method for producing banana-derived composition”.

The method for obtaining the banana-derived composition containing the physiologically active substance of the present invention as an active ingredient is not particularly limited. For example, when the banana-derived composition is obtained by extraction, hot water extraction, liquefied carbon dioxide extraction, alcohol extraction, or the like can be used as the extraction method.
Then, the banana-derived composition is obtained by adding a proteolytic enzyme to the pulp portion of banana to decompose the protein in the pulp portion of the banana by an enzymatic reaction, and then deactivating the proteolytic enzyme by heating or alcohol. It is preferably one. As the alcohol, an alcohol having 1 to 4 carbon atoms is preferable, and as the alcohol having 1 to 4 carbon atoms, those described above in "Production method of banana-derived composition" can be used, and preferable carbon number is The alcohols of 1 to 4 are the same as those described above in the "method for producing a composition derived from banana".

Furthermore, the banana-derived composition containing the physiologically active substance of the present invention as an active ingredient is preferably produced by the method for producing a banana-derived composition of the present invention. For example, when the banana-derived composition is produced by the above-described method for producing a banana-derived composition of the present invention, the physiologically active substance containing the composition A (primary purified product) has an excellent increase in blood glucose level. A physiologically active substance that has an inhibitory action and a collagen production promoting action, and further contains a composition B (secondary refined product) that has been subjected to a desalting and desalting treatment, has an excellent inhibitory action on blood sugar level elevation, a collagen production promoting action, and an anti-bacterial action. Has an oxidizing effect.

And in the physiologically active substance of the present invention, the total of the amino acid content and the tryptophan derivative in the banana-derived composition is preferably 7% by mass or more, more preferably 8% by mass or more, and particularly preferably 10% by mass or more. .. When the total of the amino acid content and the tryptophan derivative in the banana-derived composition is 7% by mass or more, the physiologically active substance of the present invention can have excellent physiological activity. When the method for producing a banana-derived composition of the present invention is used as the banana-derived composition, for example, the amino acid content and the tryptophan derivative content are adjusted by adjusting the conditions of the enzymatic reaction and subjected to the desugar desalting step (4). It can be improved. The upper limit of the total of the amino acid content and the tryptophan derivative in the banana-derived composition is not particularly limited, but is usually 95% by mass or less.
Furthermore, in the composition derived from banana, it is preferable that the amino acid contains phenylalanine and tryptophan, and the tryptophan derivative contains serotonin. When the banana-derived composition contains phenylalanine, tryptophan, and serotonin, the physiologically active substance of the present invention can have excellent physiological activity.
In the present invention, the contents of the amino acid, tryptophan derivative, phenylalanine, tryptophan, and serotonin of the banana-derived composition can be measured using the measuring method described in the examples of the present specification.

The physiologically active substance of the present invention may contain a component other than the banana-derived composition such as an excipient depending on its purpose and the like, and its use is not particularly limited, and it is used by blending in various foods and drinks. It is also possible to use it alone or as a supplement by mixing it with other components.

Hereinafter, the present invention will be specifically described based on Examples, but the present invention is not limited to these Examples. In the following description, "%" and "parts" representing amounts are based on mass unless otherwise specified.
The following methods were used to measure the Brix degree and analyze the components of the banana-derived composition.
<Brix degree measurement>
It measured at the measurement temperature of 25 degreeC using the refractometer for sugars (Koshimizu company make, sugar content meter WZ-113).
<Component analysis of banana-derived composition>
The content and type of amino acids were measured using an automatic amino acid analyzer (manufactured by JEOL Ltd., Hitachi High-Technologies Corp.). The content and type of tryptophan derivative (serotonin) were measured by an absolute calibration curve method using a reagent of the corresponding compound using high performance liquid chromatography (manufactured by Shimadzu Corporation). Then, Shodex RI-71 (manufactured by Showa Denko KK) was used to measure the content and type of sugar by an absolute calibration curve method using a putative sugar reagent.

(Example 1) Production of composition 1 derived from banana (composition A primary refined product deactivated by alcohol) Immature banana fruit (variety: Giant Cavendish Brix degree: 1.4°Bx) 196.5 parts The peel was removed to obtain 104.2 parts of the flesh. The resulting pulp portion was subdivided using a cutter mill grinder to obtain 100 parts of a paste-like pulp subdivided product.
190 parts of ion-exchanged water was added to 100 parts of the resulting pulp fragment, and the mixture was further stirred, and 5 parts of protease M "Amano" SD as a proteolytic enzyme was added to obtain a mixture. The mixture was heated to 50° C. and maintained at the temperature for 4 hours, and then further heated to 58 to 60° C. and maintained at the temperature for 2 hours to allow the enzymatic reaction to proceed. The resulting enzyme reaction product (295 parts) was left to cool to 25 to 30°C, 291.1 parts of ethanol was added thereto, and the mixture was stirred at 25 to 30°C for 2 hours to prepare an alcohol (ethanol) additive. Got The obtained alcohol additive was filtered at room temperature using a single plate filter. At this time, 29.5 parts of ethanol was further used to wash the extracted components remaining on the solid phase. By this filtration, the alcohol additive was separated into a solid phase (42.8 parts light brown) and a liquid phase (572.8 parts yellow transparent liquid). The liquid phase was concentrated under reduced pressure under vacuum while maintaining the product temperature at 40°C or lower to obtain 50 parts of concentrated liquid. This concentrated solution was pre-frozen for about 2 hours until the product temperature reached -30°C to -25°C. After that, at a product temperature of -30 to -25°C, the degree of vacuum is controlled to a pressure of 40 kPa or less under reduced pressure, and then the shelf temperature is gradually heated to 40°C. After a lapse of time, freeze-drying was terminated to obtain 11 parts of banana-derived composition 1 (yield 11% based on the pulp fragment). In this composition 1 derived from banana, fructose was 5.4%, glucose was 45.4%, sucrose was 0%, amino acid was 11.0% (phenylalanine 0.89%, tryptophan 0.34%), serotonin (tryptophan). Derivative) was 0.004%.

(Example 2) Production of banana-derived composition 2 (composition A primary purified product, alcohol-inactivated) of immature banana fruit (variety: Giant Cavendish Brix degree: 1.4°Bx) of 196.5 parts The pericarp was removed to obtain 103.2 parts of the flesh. The resulting pulp portion was subdivided using a cutter mill grinder to obtain 100 parts of a paste-like subdivided pulp.
To 100 parts of the obtained pulp fragmentation product, 200 parts of ion-exchanged water was added and stirred, and further 5 parts of protease M "Amano" SD as a proteolytic enzyme was added to obtain a mixture. The mixture was heated to 50° C. and maintained at the temperature for 4 hours, and then further heated to 60° C. and maintained at the temperature for 2 hours to allow the enzymatic reaction to proceed. The obtained enzyme reaction product (305 parts) was left to cool to 23° C., 1087.7 parts of methanol was added, and the mixture was stirred at 13 to 23° C. for 2 hours to obtain an alcohol (methanol) additive. The obtained alcohol additive was filtered at room temperature using a single plate filter. At this time, the solid phase was further washed with 14 parts of methanol to wash the extracted components remaining on the solid phase. By this filtration, the alcohol additive was separated into a solid phase (42.1 parts) and a liquid phase (after filtration, 70.2 parts of methanol was further added and 1434.4 parts of orange transparent liquid). This liquid phase was concentrated under reduced pressure under vacuum for 11 hours while maintaining the product temperature at 40° C., and further dried at 40° C. for 24 hours in a tray vacuum dryer to obtain 8.2 parts of banana-derived composition 2. Obtained (yield 8.2% based on the pulp fragment). In this banana-derived composition 2, 4.70% fructose, 43.41% glucose, 1.34% sucrose, 13.64% amino acids (phenylalanine 1.13%, tryptophan 0.42%), serotonin ( Tryptophan derivative) was 0.0009%.

(Example 3) Production of banana-derived composition 3 (composition B secondary refined product deactivated by alcohol) As described in "(Example 1) Production of banana-derived composition 1", the pulp portion of an unripe banana was prepared. It was subdivided, enzymatically reacted, filtered, and concentrated to obtain 50 kg of concentrated liquid (containing 11.0 kg of solid content). To this concentrated liquid, 150 kg of ion-exchanged water was added to obtain 200 kg of a diluted liquid (solid content concentration 5.5%).
200 kg (200 L, 20 RV) of this diluted solution was passed through a column filled with 10 L (1 RV) of a synthetic adsorbent (Separ beads (registered trademark) SP207 manufactured by Mitsubishi Chemical Corporation) at a flow rate of 15 L/h (1.5 SV). Then, 80 L (8 RV) of ion-exchanged water was passed through at the same flow rate and washed with water. The Brix degree of the ion-exchanged water after passing through the column was measured immediately before the completion of passing the whole amount of the ion-exchanged water, and it was confirmed that the Brix degree was 0.
Then, 180 L (18 RV) of 5 mass% ethanol aqueous solution was passed at the same flow rate, and then 180 L (18 RV) of 7 mass% ethanol aqueous solution was passed at the same flow rate, and the liquid (360 L) after passing was recovered. .. The concentration of tryptophan contained in the liquid after passing through the column was measured by high-performance liquid chromatography manufactured by Shimadzu Corporation immediately before the completion of passing the whole amount of the 7 mass% ethanol aqueous solution, and it was confirmed that the concentration was 0.
After passing through the liquid, 360 L of the liquid was concentrated under reduced pressure under vacuum while maintaining the product temperature at 40°C or lower to obtain 0.83 kg of concentrated liquid. This concentrated solution was pre-frozen for about 2 hours until the product temperature reached -30°C to -25°C. After that, at a product temperature of -30 to -25°C, the degree of vacuum is controlled to a pressure of 40 kPa or less under reduced pressure, and then gradually heated to a shelf temperature of 40°C, and the product temperature becomes about the same as the shelf temperature, and 3 hours or more has passed After that, freeze-drying was terminated to obtain 0.17 kg of banana-derived composition 3 (the yield was 1.5% based on 11.0 kg of the solid content of the initially concentrated liquid). In this banana-derived composition 3, fructose was 0%, glucose was 0%, sucrose was 0%, phenylalanine was 3.7%, tryptophan was 11.8%, and serotonin (tryptophan derivative) was 0.15%. ..

(Example 4) Production of banana-derived composition 4 (composition B secondary refined product deactivated by alcohol) As described above in "(Example 2) Production of banana-derived composition", the pulp portion of the unripe banana was subdivided. It was lysed, subjected to enzymatic reaction, filtered, and concentrated to obtain 3.5 kg of concentrated liquid (containing 0.8 kg of solid content and 23% solid content concentration).
3.5 L (3.5 L, 3.5 RV) of this concentrated liquid was charged in a column packed with 1 L (1 RV) of a strongly acidic cation exchange resin (DIAION (registered trademark) SK1B manufactured by Mitsubishi Chemical Co., Ltd.) at 1.5 L/ The liquid was passed through at a flow rate of h (1.5 SV), and then 15.4 L (15.4 RV) of ion-exchanged water was passed through at the same flow rate. The Brix degree of the ion-exchanged water after passing through the column was measured immediately before the completion of passing the whole amount of the ion-exchanged water, and it was confirmed that the Brix degree was 0.
Then, 10 L (10 RV) of 1 mol/L ammonia water was passed at the same flow rate, and the liquid (10 L) after passing was recovered. The concentration of tryptophan contained in the eluent after passing through the column was measured by high performance liquid chromatography (manufactured by Shimadzu Corporation) immediately before the completion of passing the whole amount of 1 mol/L ammonia water, and it was confirmed that the concentration was 0.
After passing through the liquid, 15.4 L of liquid was concentrated under reduced pressure under vacuum while maintaining the product temperature at 40° C. or lower to obtain 0.70 kg of concentrated liquid. This concentrate is pre-frozen for about 2 hours until the product temperature reaches -30°C to -25°C, and then the product temperature is controlled at -30 to -25°C under a reduced pressure to a vacuum degree of 40 kPa or less, and then gradually. After heating to a shelf temperature of 40° C., and after the product temperature reached the shelf temperature and became equal to the shelf temperature for 3 hours or more, lyophilization was terminated to obtain 0.16 kg of banana-derived composition 4 (initial concentration The yield based on the solid content of the liquid of 0.8 kg is 20%). In this banana-derived composition 4, fructose was 0%, glucose was 0%, sucrose was 0%, amino acids were 55.2% (phenylalanine 4.47%, tryptophan 1.69%), and serotonin (tryptophan derivative) was 0. It was 0.015%. In addition, when the potassium content in this banana-derived composition 4 was measured, it was found to be 7 mg/100 g, which was extremely low, indicating that desalination was successful.

(Comparative Example 1) Production of banana-derived composition 5 1.63 parts of banana-derived composition 5 was obtained in the same manner as in Example 1 except that enzymatic decomposition was not performed. This yield is extremely lower than that of the banana-derived composition 1 of Example 1, and it is understood that the yield of the banana-derived composition can be dramatically improved by enzymatic decomposition.
In this banana-derived composition 5, fructose was 16.3%, glucose was 25.6%, sucrose was 0%, amino acids were 6.6% (phenylalanine 0.3%, tryptophan 0.102%), serotonin. (Tryptophan derivative) was 0.009%. Thus, the content of the active ingredient is extremely lower than that of the banana-derived composition 1 of Example 1, and even with the banana-derived composition 5, sufficient blood glucose elevation suppression and collagen production promoting physiological activity can be obtained. I can't.

Example 5 Production of Banana-Derived Composition 6 (Composition A Primary Purified Product Heat Inactivated) Immature banana fruit (variety: Giant Cavendish Brix degree: 1.4° Bx) of 196.5 parts The peel was removed to obtain 104.2 parts of the flesh. The obtained pulp portion was subdivided using a cutter mill grinder to obtain 100 parts of a paste-like subdivided pulp portion.
190 parts of ion-exchanged water was added to 100 parts of the resulting pulp fragment, and the mixture was further stirred, and 5 parts of protease M "Amano" SD as a proteolytic enzyme was added to obtain a mixture. The mixture was heated to 50° C. and maintained at the temperature for 4 hours, and then further heated to 60° C. and maintained at the temperature for 15 hours to allow the enzymatic reaction to proceed. The obtained enzyme reaction product (295 parts) was filtered through Celite, and further filtered using a filter having a pore size of 0.45 μm. The proteolytic enzyme was inactivated by heating the obtained filtrate at 90°C for 30 minutes, and then left standing and cooled to 25 to 30°C to obtain an enzyme reaction product. The enzyme reaction product was concentrated under reduced pressure under vacuum while maintaining the product temperature at 40° C. or lower to give a solid content concentration of about 10 mass %. To the enzyme reaction product after concentration under reduced pressure, as an excipient, Paindex (registered trademark) #100 in the same amount as the solid content mass of the enzyme reaction product was added, and the mixture was sterilized with stirring at 90° C. for 1 hour. After stirring and sterilization, the mixture was allowed to stand and cooled to 25 to 30° C., and then freeze-dried to obtain 16.8 parts of banana-derived composition 6.

<Confirmation of blood glucose elevation inhibitory effect (rat)>
A 7-week-old male Wistar rat was fasted for 24 hours, and thereafter, 10 mg of a banana-derived composition 2 (primary purified product) as a physiologically active substance and 2 mL of water were added to the stomach of the rat (N=5) per 1 g of the rat body weight. The mixture was directly administered using a gastric tube. Before, 15 minutes, 30 minutes, 45 minutes, and 60 minutes after the administration of the banana-derived composition 2, blood was collected from the tail vein of the rat, and the blood glucose level was measured using Terumo Medisafe Fit. All the operations were performed under anesthesia (note that it was separately confirmed that there was no effect of anesthesia on blood glucose level).
Then, after 1 week, the same rat (N=5) used was replaced with the banana-derived composition 2 and the same amount of sugars (fructose and glucose) contained in the banana-derived composition 2 was dissolved in water. , Was similarly administered as a placebo. Before, 15 minutes, 30 minutes, 45 minutes, and 60 minutes after the administration of the placebo, blood was collected from the rat tail vein and the blood glucose level was measured. All operations were performed under anesthesia.
Further, another 7-week-old male Wistar rat (N=5) was first subjected to a test using the same placebo as above, and the blood glucose level was measured in the same manner as above. Then, one week later, a test using the composition 2 derived from banana was conducted in the same manner as above, and the blood glucose level was measured.
When the banana-derived composition 2 was administered and the placebo was administered, changes in blood glucose level with time are shown in FIG. 1 (each average value of N=10). From FIG. 1, it can be seen that the banana-derived composition 2 (primary purified product) as a physiologically active substance exerts an excellent effect of suppressing an increase in blood glucose level.

<Confirmation of blood sugar elevation inhibitory effect (human)>
The test subjects (healthy Japanese males aged 20 to 70) completed the dinner the day before the test by 21:00, and then fasted (only water was ingestible) and tested (placebo, banana-derived composition 6). ) Was taken 1 hour before the time (about 9 am) to complete fasting until all blood collection on the day was completed.
First, a test was conducted using a placebo. The test subject separately mixed 12.8 g of placebo (15 g of banana-derived composition 6 with glucose, fructose, and maltitol as the excipient in the same amount, separately mixed with glucose, fructose, and maltitol). Oral ingestion of the mixture prepared in step 2) with 200 mL of water, and then the load meal (300 g of cooked rice, Kukukure curry (registered trademark) medium spiciness, 180 mL of water) is divided into 3 parts, and each is ingested for 5 minutes as a guide. Then, the whole amount was ingested over 15 minutes. Blood was collected after the placebo was ingested, before the loading meal was ingested, and 30, 60, and 90 minutes after the completion of the loading meal, and the blood glucose level and insulin were measured.
Next, a test was conducted using the banana-derived composition 6 four days after the test date of the test using the placebo. The subject orally ingested 15 g of the banana-derived composition 6 together with 200 mL of water, then divided the above-mentioned loaded meal into 3 parts, ingested each for 5 minutes as a standard, and ingested the entire amount over 15 minutes. .. Blood was collected after ingestion of the banana-derived composition 6 before ingestion of the loaded diet, and at 30, 60, and 90 minutes after the end of ingestion of the loaded diet, and the blood glucose level and insulin were measured. Two days later, the same operation as above was performed except that the amount of the banana-derived composition 6 was 7.5 g, and two days later, except that the amount of the banana-derived composition 6 was 3.75 g. The same operation as described above was performed.
When the banana-derived composition 6 was administered, changes in blood glucose level and insulin with the passage of time when a placebo was administered were calculated using the index values with the value before ingestion of the loaded diet as 100, respectively, and FIG. As shown in FIG. From FIG. 2 and FIG. 3, it can be seen that the banana-derived composition 6 (primary purified product) as a physiologically active substance exerts an excellent effect of suppressing an increase in blood glucose level.

<Confirmation of collagen production promoting action>
Normal human dermal fibroblasts were seeded in a 24-well culture plate at a concentration of 2×10 ⁴ cells/ml and cultured in a CO ₂ incubator at 37° C. for 72 hours. As the culture medium, DMEM medium (manufactured by SIGMA) containing 5% fetal bovine serum (manufactured by Thermo Trace) was used.
After removing the pre-culture medium, 1 mL of full-scale culture medium was prepared per well on a 24-well culture plate. As the main culture medium, DMEM medium (manufactured by SIGMA) containing 0.25% fetal bovine serum (manufactured by Thermo Trace) was used. Twenty-four hours after the replacement with the main culture medium, the same main culture medium was replaced again and the sample was added. As samples, control (distilled water), banana-derived composition 2 (addition concentration 100 μg/ml), banana-derived composition 2 (addition concentration 200 μg/ml) were used (test samples were prepared using distilled water, Filter sterilized with 0.22 μm filter).
After each sample was added, it was cultured in a CO ₂ incubator at 37° C. for 72 hours, and the production amount of procollagen after the culture was measured. Specifically, using the Procollagen Type I C-Peptide (RIP) EIA Kit (manufactured by Takara Bio Inc.) capable of quantifying Procollagen Type I C-Peptide (PIP), the production amount of type I procollagen in the culture supernatant was measured. It was measured. The cell number was determined using Cell counting Kit-8 (manufactured by Dojindo Laboratories).
As a sample, the amount of procollagen (Procollagen) produced when the control was added and when the composition 2 derived from banana was added is shown in FIG. 4 (each average value of N=4, error bars represent actual measurement results). The range is shown and the control is shown as 100%). From FIG. 4, it can be seen that the banana-derived composition 2 (primary purified product) as the physiologically active substance has an excellent collagen production promoting action.

<Confirmation of antioxidant effect>
Banana-derived composition 4 (secondary purified product) as a physiologically active substance and vitamin C are dissolved in ultrapure water to prepare a solution having a concentration of 10 mg/ml, and the antioxidant ability of each of these solutions is measured twice. did. Specifically, the antioxidative ability against the oxidation of hypozinc acid (HClO) was measured by an OXY adsorption test using a free radical analyzer FREE (manufactured by Wismer). As a result of the measurement, the erased HClO concentration is obtained in the unit of μmol HClO/mL. The higher this number is, the more HClO is eliminated, that is, the higher antioxidative effect is. The results are shown in Table 1.

From Table 1, it can be seen that the banana-derived composition 4 (secondary purified product) as a physiologically active substance has a higher μmol HClO/mL value than vitamin C, that is, exhibits a high antioxidant effect.

<Confirmation of filtration (solid-liquid separation)>
Unripe banana pulp (variety: Giant Cavendish Brix degree: 4.5°Bx), ripe banana pulp part (variety: Giant Cavendish Brix degree: 24.8°Bx), raw materials In order to confirm the filterability when used for, a comparative test was conducted in the following manner.
In the same manner as in the method described in "(Example 1) Production of banana-derived composition 1", the pulp portion of the above-mentioned unripe banana was subdivided, and the obtained subdivided pulp (150 g) was subjected to an enzymatic reaction to obtain an enzymatic decomposition product. Got
This enzymatic degradation product was used as a filtration test sample 1, and ethanol was added to the filtration test sample 1 so that the ethanol concentrations were 10%, 30%, and 50%, respectively. And
On the other hand, in the same manner as the above-mentioned unripe banana, the pulp portion of the above-mentioned ripe banana was subdivided, and the obtained subdivided pulp (150 g) was subjected to an enzymatic reaction to obtain an enzymatic decomposition product.
This enzymatic degradation product was used as sample 5 for filtration test, and ethanol added to sample 5 for filtration test to have ethanol concentrations of 10% and 50%, respectively, were samples 6 and 7 for filterability test.
Samples 1 to 7 for filtration test were suction-filtered using qualitative filter paper No. 2 (manufactured by ADVANTEC), and the time required for complete filtration was visually confirmed, and the time required for total filtration (filtration time ) Was measured. The results are shown in Table 2.

From Table 2, it can be seen that the case where the pulp portion of the unripe banana is used has extremely excellent filterability as compared with the case where the pulp portion of the ripe banana is used. In addition, when the pulp part of an unripe banana is used, it is understood that the filterability can be improved as the ethanol concentration is increased.

<Confirmation of extraction efficiency due to difference in alcohol used for deactivation>
In order to confirm the superiority or inferiority of the extraction efficiency of the amino acid and tryptophan derivative due to the difference in the alcohol having 1 to 4 carbon atoms used for deactivating the protease, a comparative test was conducted in the following manner.
Banana-derived composition 1 of Example 1 using ethanol, Example 1 and three kinds of bananas obtained in the same manner except that methanol was used instead of ethanol, 2-propanol, and 1-butanol, respectively. The amounts of phenylalanine, tryptophan, and serotonin contained in the derived composition were measured. The results are shown in Table 3.

From Table 3, it can be seen that ethanol and methanol can efficiently extract the above-mentioned components in the banana pulp part as a whole more efficiently than 2-propanol and 1-butanol.

ADVANTAGE OF THE INVENTION According to this invention, the manufacturing method of the composition derived from a banana which can obtain the composition which shows the outstanding physiological activity from the pulp part of a banana can be provided.
Further, according to the present invention, it is possible to provide a physiologically active substance containing a banana-derived composition derived from the pulp portion of banana, which exhibits excellent physiological activity.

Claims (8)

A bioactive substance containing a banana-derived composition obtained from the pulp part of a banana as an active ingredient ,
The sum of the amino acid content and the tryptophan derivative content in the banana-derived composition is 10% by mass or more and 95% by mass or less,
The physiologically active substance, wherein the amino acid contains phenylalanine and tryptophan, and the tryptophan derivative contains serotonin. The physiologically active substance according to claim 1, wherein the banana-derived composition has an amino acid content of 11.0% by mass or more and 55.2% by mass or less. The physiologically active substance according to claim 1 or 2, wherein the content of the phenylalanine in the banana-derived composition is 0.89% by mass or more and 4.47% by mass or less. The physiologically active substance according to any one of claims 1 to 3, wherein a content of the tryptophan in the banana-derived composition is 0.34% by mass or more and 11.8% by mass or less. The physiologically active substance according to any one of claims 1 to 4, wherein the content of the serotonin in the banana-derived composition is 0.0009% by mass or more and 0.15% by mass or less. The content of the phenylalanine in the banana-derived composition is 0.89% by mass or more and 4.47% by mass or less, the content of the tryptophan is 0.34% by mass or more and 11.8% by mass or less, and The physiologically active substance according to claim 1 or 2, wherein the content of serotonin is 0.0009% by mass or more and 0.15% by mass or less. The physiologically active substance according to any one of claims 1 to 6 , which exerts an action of suppressing an increase in blood glucose level. The physiologically active substance according to any one of claims 1 to 7 , which exhibits a collagen production promoting action.

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