JP6055874B2 - Method of using citric acid, food / beverage product composition for enzyme activation, composition for enzyme activation, and agent for enzyme activation - Google Patents

Description

The present invention is the use of click-ene acids, enzyme activation for food or beverage composition, the enzyme activating compositions and enzyme activating agent.

  In general, citric acid is abundant in citrus fruits such as lemon and tangerine (for example, 1350 mg / one lemon), and is known as a substance that forms one component of the citric acid circuit (TCA circuit, Krebs circuit) in vivo. Yes. In addition, it has been reported that citric acid has various effects on living bodies. For example, Patent Documents 1 to 3 disclose that ingesting citric acid provides a preventive effect and an improvement / recovery promoting effect on physical and mental fatigue.

JP-A-9-059161 Japanese Patent Laid-Open No. 10-175856 JP 2000-333651 A

The purpose of this invention is the use of new citric acids, enzyme activation for food or beverage composition, the enzyme activating composition, and to provide an enzyme activating agent.

The method of using citric acid according to claim 1 is characterized in that citrate synthetase and succinate dehydrogenase are added to the composition by incorporating at least one citric acid of citric acid and citrate into the composition. A function of increasing the maximum value of the production amount per unit time of the product produced by at least one kind is provided.

The food / beverage composition composition for enzyme activation according to claim 2 , comprising at least one of citric acid and citrate as an active ingredient, and per unit time of a product produced by at least one of citrate synthetase and succinate dehydrogenase. The maximum value of the generation amount is increased.
The composition for enzyme activation according to claim 3 , comprising at least one of citric acid and citrate as an active ingredient, and a production amount of a product produced by at least one of citrate synthetase and succinate dehydrogenase per unit time. The maximum value of is increased.
The enzyme activation agent according to claim 4 comprises at least one of citrate and citrate as an active ingredient, and the amount of product produced per unit time of a product produced by at least one of citrate synthetase and succinate dehydrogenase. It is characterized by increasing the maximum value.

According to the present invention, at least one enzyme of citrate synthetase and succinate dehydrogenase can be activated. Also provided are methods for using novel citric acids .

The graph which shows the result of having measured the activity of the citrate synthetase, the succinate dehydrogenase, and the malate dehydrogenase when a citrate cycle activator is administered. The graph which shows the result of having measured the swimming duration in the case of administering a citrate circuit activator.

Hereinafter, embodiments of a citric acid cycle activator embodying the present invention will be described in detail.
The citric acid cycle activator of the present embodiment contains citric acid as an active ingredient and has an action of activating the citric acid cycle in the body. More specifically, by activating an enzyme involved in the citrate cycle, the activity of the entire citrate cycle is increased. In addition, the activation of an enzyme here includes all when the quantity of the product produced | generated per unit time increases. Factors that increase the amount of product produced by the enzyme reaction include, for example, an increase in the amount of enzyme and an increase in the reaction rate, and it is particularly preferable that the reaction rate of the enzyme is increased.

The citric acid cycle is a common pathway for the oxidative metabolism of most organisms. It oxidizes the acetyl group of acetyl-CoA (acetyl coenzyme A) to two molecules of CO ₂ and liberates NADH (nicotine). This is a system that utilizes amide adenine dinucleotide) for the production of ATP (adenosine triphosphate) in an electron transport system. Since ATP is a molecule having an action of storing and supplying energy in the living body, it can be said that the citric acid circuit plays a very important role in energy production in the living body.

Specifically, the citrate cycle involves eight enzymes: citrate synthetase, aconite hydratase, isocitrate dehydrogenase, α-ketoglutarate dehydrogenase, succinyl-CoA synthetase, succinate dehydrogenase, fumarase and malate dehydrogenase. It is configured. By the enzyme reaction of these eight enzymes, one acetyl group is oxidized to two molecules of CO ₂ , and three molecules of NADH, one molecule of FADH ₂ , and one molecule of GTP (guanosine triphosphate) or ATP are converted into Produced. The 3 molecules of NADH and 1 molecule of FADH ₂ produced here contribute to the production of 11 molecules of ATP in the electron transport system. Therefore, a total of 12 molecules of ATP are produced by one rotation of the citric acid circuit.

  Therefore, when the citric acid circuit activator of the present embodiment having an action of activating the citric acid circuit is applied to a living body, an excellent effect that energy is produced more efficiently in the living body is exhibited. In addition, since it is suggested that the function of the citric acid cycle decreases with aging, the citric acid cycle activator of the present embodiment is also useful for prevention / improvement of such functional deterioration of the citric acid cycle. Can be expected.

  Citric acids contained as an active ingredient of the citric acid cycle activator of the present invention include at least one selected from citric acid and citrate. As citric acid, any of citric anhydride and citric acid hydrate may be used. The citrate may be any salt that is pharmaceutically acceptable, and examples thereof include sodium citrate, potassium citrate, magnesium citrate, and calcium citrate. As these citric acids, any of biosynthetic products obtained by fermentation, chemically synthesized products, and extracts extracted from natural products using water, hydrophilic organic solvents, or the like may be used. As a form of citric acid compounding, a biosynthetic product, a chemically synthesized product, an extract from a natural product or a purified product may be blended directly into a citric acid circuit activator, and a material containing citric acid is used as a citric acid circuit You may mix | blend in an activator. In addition, these citric acids may be mix | blended independently and may be mix | blended in combination of 2 or more types.

  Examples of the material containing citric acids include vegetables, fruits, and moromi vinegar. In particular, fruits such as ume and citrus are rich in citric acids, and citrus fruits such as lemon, orange, lime, grapefruit and sudachi are preferably used as materials containing citric acids. When a citrus-derived raw material is used as a material containing citric acids, a compressed liquid such as citrus juice or a concentrate thereof is preferably used. In addition, you may use the pressing liquid of a fruit (fruit peel (albedo, flavedo), an agate capsule, a saona etc.), or the thing containing a part of the structural component of the fruit.

  The citric acid cycle activator of the present embodiment is mainly a pharmaceutical, quasi-drug, health food, food for specified health use, health drink, nutritional supplement, etc. that has the effect of activating the citric acid cycle. It is ingested by blending into the composition.

  When the citric acid cycle activator of the present embodiment is used as a pharmaceutical, it is possible to adopt any administration method such as intravascular administration or transdermal administration, in addition to administration by taking (oral intake). . Although it does not specifically limit as a dosage form, For example, a powder, a powder agent, a granule, a tablet, a capsule, a pill, a suppository, a liquid agent, an injection, etc. are mentioned. In addition, excipients, bases, emulsifiers, solvents, stabilizers and the like as additives may be blended as long as the object of the present invention is not impaired.

  When using the citric acid cycle activator of the present embodiment as a food or drink, by adding it to various food materials or beverage materials, for example, powder, tablet, granule, liquid (drink agent, etc.) It can be processed into capsules, syrups, candies and the like and used as health food preparations, dietary supplements and the like. Specific examples of the above-mentioned food and drink include sports drinks, tea beverages extracted from tea leaves and herbs, dairy products such as milk and yogurt, foods containing gelling agents such as pectin and carrageenan, glucose, sucrose, and fructose. And beverages and foods containing sugars such as lactose and dextrin, flavorings, sweeteners such as stevia, aspartame, sugar alcohols, and fats and oils such as vegetable oils and animal fats. In addition, a substrate, an excipient, an additive, a secondary material, a bulking agent, and the like may be added as appropriate within a range that does not impair the object of the present invention.

  In addition, when ingesting this citric acid cycle activator as a pharmaceutical or food and drink, the daily citric acid content for adults is preferably 1 to 10 g, more preferably 2 to 5 g. When the daily intake of citric acids, which are active ingredients, is less than 1 g, the citric acid cycle activation action by citric acids may not be effectively enhanced. On the other hand, if the daily intake exceeds 10 g, it is uneconomical because much of it is excreted.

Next, operational effects in the present embodiment will be described below.
(1) The citric acid cycle activator of this embodiment has an action of activating citric acid circuits in the body using citric acids as active ingredients. For this reason, the citric acid circuit activator of this embodiment can be used as a biological function promoter that promotes efficient energy production in vivo. In addition, the citric acid cycle activator of the present embodiment is also useful for prevention / improvement of functional deterioration of the citric acid cycle. In addition, by improving the efficiency of energy production in the living body, it is possible to improve exercise endurance and reduce fatigue.

  (2) Since citric acids in the present embodiment are contained in a large amount in citrus fruits, the citric acids can be purified and obtained easily and appropriately by performing an extraction treatment or the like. In particular, when lemon is used as a citrus fruit, it is possible to easily obtain a high concentration of citric acid. In addition, when citric acids derived from citrus fruits are used, they are derived from natural ingredients, so they are highly safe and can be easily applied to pharmaceuticals and foods and drinks.

In addition, you may change the said embodiment as follows.
-You may use the citric acid cycle activator of this embodiment for animals other than humans, such as a horse and a cow, for example.

-You may use the citrate circuit activator of this embodiment as an enzyme activator which activates the enzyme involved in a citrate circuit.
-You may use the citric acid circuit activator of this embodiment as a fatigue reducing agent.

Next, the embodiment will be described in more detail with reference to each test example.
(Test Example 1: Test on activation of enzymes involved in citrate cycle by citrate cycle activator)
A plurality of ddY mice (6 weeks old, male) were prepared, and these were subjected to swimming exercise after 5 days of preliminary breeding, and 2 of administration group 1 and control group 1 so that body weight and swimming time were averaged. They were grouped into groups, and these were used to verify the citric acid cycle activation effect of the citric acid cycle activator of the present invention. In the ddY mice in the administration group, citric acid in an amount of 125 mg / kg body weight was dissolved in 500 μl of distilled water (Example 1), and in the ddY mice in the control group, 500 μl of distilled water (Comparative Example 1). ) Was orally administered periodically. The administration schedule was 6 days a week (Monday to Saturday), once a day, in principle between 16:00 and 17:00, and continued for 5 weeks. Each group of ddY mice was allowed to perform a swimming exercise every week. The swimming exercise was performed until a weight of 7% of the body weight was applied and the head was submerged under the surface of the water for 5 seconds.

  On week 5 of administration, ddY mice of each group were dissected without performing swimming exercise. Then, the activities of citrate synthetase, succinate dehydrogenase and malate dehydrogenase in the muscle of each group of ddY mice were measured.

  Citrate synthase activity was measured by the method of Srere et al. Specifically, 0.1 M Tris-HCl Buffer containing 1 mM DTNB and 10 mM acetate-CoA was added to the muscle homogenate solution, left in warm water at 25 ° C. for 10 minutes, and after reaching temperature equilibrium, 10 mM oxaloacetate was added. The reaction was initiated by addition. The change in absorbance at 412 nm was measured using a spectrophotometer for 5 minutes after the start of the reaction.

  Succinate dehydrogenase activity was measured by the method of Acklel et al. Specifically, 0.3 M phosphate buffer, 30 mM EDTA, 0.4 M succinic acid, 3% bovine serum albumin, 10 mM potassium ferricyanide were added, and the mixture was allowed to stand in warm water at 25 ° C. for 10 minutes. Muscle homogenate solution was added to initiate the reaction. The change in absorbance at 455 nm for 10 minutes after the start of the reaction was measured.

  The malate dehydrogenase activity was measured by the method of Pattak et al. Specifically, the muscle homogenate solution was diluted with 500 mM imidazole-HCl buffer pH 7.4. 0.02 ml thereof was taken into a microcell, 50 mM imidazole-HCl buffer pH 7.4 was added, 1.5 mM NADH was added, and the mixture was allowed to stand at 30 ° C. for 5 minutes, and the absorbance was measured at 412 nm. Furthermore, 5 mM oxaloacetate was added, and the mixture was allowed to stand for 1 minute in the same manner, and absorbance was measured for 3 minutes.

  For the significant difference test, a corresponding t-test was performed for comparison between the administration group 1 administered with Example 1 and the control group 1 administered with Comparative Example 1. The results are shown in FIG. In FIG. 1, “*” indicates that the result of the t test is p <0.05.

  From the results shown in FIG. 1, the activity of any of citrate synthetase, succinate dehydrogenase and malate dehydrogenase in the administration group 1 administered with Example 1 compared to the control group 1 administered with Comparative Example 1. It was confirmed that there is a tendency to increase. In particular, it was confirmed that the enzyme activities of citrate synthetase and succinate dehydrogenase were significantly increased when Example 1 was administered compared to the case where Comparative Example 1 was administered.

  From the above results, it was confirmed that citrate synthetase, succinate dehydrogenase and malate dehydrogenase activities were increased by ingesting citric acid. Since citrate synthetase, succinate dehydrogenase and malate dehydrogenase are enzymes involved in the citrate cycle, it can be said that ingesting citrate increases the activity of the entire citrate cycle.

  As described above, since the citric acid circuit is a system that contributes to energy production in the living body, it is suggested that when the citric acid circuit is activated, energy is more efficiently produced in the living body. The From these results, it can be said that the citric acid cycle activator of Example 1 exhibits an effect of promoting efficient energy production in vivo. Therefore, it can be used as a biological function promoter that promotes efficient energy production in vivo.

(Test Example 2: Test for improving exercise endurance by citric acid cycle activator)
A plurality of ddY mice (4 weeks old, male) were prepared, and these were subjected to swimming exercise after preliminary breeding for 7 days so that the body weight and the swimming time were averaged. The group 2 was divided into three groups, and the effects of improving the exercise endurance of the citric acid cycle activator of the present invention were verified using these groups. In the ddY mice in the administration group 2, citric acid in an amount of 62.5 mg / kg body weight was dissolved in 500 μl of distilled water (Example 2), and in the ddY mice in the administration group 3 125 mg / kg body weight. A solution (Example 3) in which an amount of citric acid to be dissolved was dissolved in 500 μl of distilled water, and 500 μl of distilled water (Comparative Example 2) was periodically orally administered to the ddY mice of Control Group 2. Examples 2, 3 and Comparative Example 2 were administered once a day and continued for 5 weeks. In addition, swimming exercise was performed every week, and swimming duration was measured. The swimming duration was defined as the time required for the head to sink for 5 seconds under the water surface in a swimming exercise with a weight of 10% of the body weight.

  As for the significant difference test, a corresponding t test was performed for comparison between the administration groups 2 and 3 to which Examples 2 and 3 were administered and the control group 2 to which Comparative Example 2 was administered. The results are shown in FIG. In FIG. 2, “**” indicates that the result of the t-test is p <0.01, and “***” indicates that the result of the t-test is p <0.005. .

  From the results of FIG. 2, it was confirmed that the swimming duration was increased in the administration groups 2 and 3 to which Examples 2 and 3 were administered, as compared with the control group 2 to which Comparative Example 2 was administered. In particular, it was revealed that the swimming duration of the administration group 3 to which Example 3 was administered was significantly longer than that of the control group 2 after the third week. This improvement in exercise endurance is thought to be due to the citric acid cycle being activated by ingesting citric acid. This result also suggests that citric acid has anti-fatigue action.

Next, a technical idea that can be grasped from the above embodiment and another example will be described.
○ An enzyme activator comprising citric acid as an active ingredient and activating an enzyme involved in the citric acid cycle.

A citrate synthetase activator comprising citrates as an active ingredient and activating citrate synthetase.
A succinate dehydrogenase activator characterized by containing citric acid as an active ingredient and activating succinate dehydrogenase.

O A malate dehydrogenase activator comprising citrates as an active ingredient and activating malate dehydrogenase.
○ The citric acid cycle activator, wherein the citric acid is derived from citrus fruits.

  A beverage composition comprising the citric acid cycle activator as an active ingredient.

Claims (4)

  By adding at least one citric acid of citric acid and citrate into the composition, a product produced by at least one of citrate synthetase and succinate dehydrogenase per unit time is produced for the composition. A method for using citric acids, which is provided with a function of increasing the maximum amount.   Enzyme activation characterized by increasing the maximum production amount per unit time of a product produced by at least one of citrate and citrate and at least one of citrate synthetase and succinate dehydrogenase Food and beverage composition.   Enzyme activation characterized by increasing the maximum production amount per unit time of a product produced by at least one of citrate and citrate and at least one of citrate synthetase and succinate dehydrogenase Composition.   Enzyme activation characterized by increasing the maximum production amount per unit time of a product produced by at least one of citrate and citrate and at least one of citrate synthetase and succinate dehydrogenase Agent.