JP2008208104A - Antioxidants and food and drink - Google Patents

Abstract

The present invention provides an antioxidant and a food and drink containing the same, which have a higher antioxidant action and can simplify the production process.
An antioxidant is a culture obtained by culturing mycelia of basidiomycetes and / or ascomycetes in a medium prepared from a raw material containing algae, or an extract prepared from the culture. Is contained as an active ingredient. As algae, chlorella and / or spirulina are preferable, and as basidiomycetes and / or ascomycetes, shiitake mushrooms or mannen moths are preferable. The culture is preferably collected as an extract obtained by self-digestion with an enzyme contained in the mycelium of basidiomycetes and / or ascomycetes. This antioxidant can also be added to various foods and drinks.
[Selection figure] None

Description

  The present invention relates to an antioxidant and a food and drink obtained by culturing mycelium of basidiomycetes and / or ascomycetes using a medium containing algae.

Animals are aerobic organisms that use oxygen to gain energy. Thus, oxygen is essential for survival, but active oxygen is produced as a byproduct of oxygen utilization. As reactive oxygen species, superoxide anion radical (O ₂ ⁻ ), hydrogen peroxide (H ₂ O ₂ ), hydroxy radical (.OH), singlet oxygen ( ¹ O ₂ ), lipid radical (RO.), Peroxide (ROOH), peroxy radical (ROO.), Nitric oxide (NO), and the like are known.

  Certain kinds of active oxygen were produced as weapons to attack foreign bodies from immune system cells such as lymphocytes and macrophages, and because of their high killing ability, they were infected with bacteria and viruses that entered the body. It is useful for attacking objects (foreign substances) that are undesirable for a living body such as cells and cancerous cells, protecting the living body from external enemies, and maintaining homeostasis.

  However, the high reactivity of active oxygen and the strength of killing power not only attack foreign substances, but also attack and damage normal proteins, lipids, nucleic acids and the like in the living body. For this reason, surplus active oxygen attacks the body, causing heart disease such as arteriosclerosis and myocardial infarction, cerebral infarction, cerebral hyperemia, diabetes, kidney damage, cancer, Alzheimer's dementia, Parkinson's disease, etc. It is also a cause.

  The production amount of these active oxygen is increased by radiation, ultraviolet rays, smoking, intense exercise and stress. The modern society is a stress society where complexity and efficiency are required, and there is an overproduction of active oxygen, such as an increase in the amount of ultraviolet rays due to environmental destruction, radiation exposure for diagnosis, and flooding of chemical substances.

  In order to protect yourself from active oxygen, it is necessary to quickly remove excess active oxygen. In order to perform this function, the living body is equipped with enzymes such as superoxide dismutase (SOD), catalase, and glutathione peroxidase.

  In addition to enzymes, substances that eliminate active oxygen are known. These are fat-soluble antioxidants such as vitamin A, vitamin E, β-carotene, and lycopene, and water-soluble antioxidants such as vitamin C, catechin, polyphenols such as flavonoids and anthocyanins, In a healthy state, the living body is protected from the damage of active oxygen by the action of an enzyme that scavenges active oxygen and the action of a substance that scavenges active oxygen.

  Unfortunately, however, enzymes that scavenge active oxygen are known to decrease with aging and stress. In addition, in the current dietary life, convenience, convenience, and deliciousness are prioritized, so food with sufficient antioxidant effect is not consumed.

  For this reason, injury due to active oxygen accumulates as the population ages, and the occurrence of life-threatening illnesses such as arteriosclerosis, hypertension, myocardial infarction, angina pectoris, cerebral infarction, diabetes, and cancer becomes a problem. In addition, even young people are suffering from active oxygen injury due to uneven eating habits, and an increasing number of people have become so-called metabolic syndrome or their reserves.

  Under such circumstances, various foods containing antioxidants extracted from natural raw materials have been proposed. For example, in Patent Document 1 below, the cyanobacterium Spirulina, Nenjumo, or Yuremo, It contains at least one extract of a microalgae selected from the genus Chloodactilon, Rhodospora, or Crooteke, or the genus Pleurocrisis of the haptoalgae, or the primordiale Phaeocystis. Furthermore, foods characterized by having an antioxidant action are disclosed.

  Patent Document 2 below discloses an antioxidant characterized by containing chlorella heat-treated at normal pressure or under pressure as an active ingredient.

  Furthermore, the following Patent Document 3 contains an antioxidant as an active ingredient containing two kinds of mixtures selected from the group consisting of agaricus, mesimacob, propolis, and chlorella heat-treated at normal pressure or under pressure. Is disclosed.

Furthermore, in Patent Document 4 below, a mycelium of basidiomycetes and / or ascomycetes is cultured using a medium containing a plant fiber component, and a saccharide, a protein, a water solution obtained by extraction from this culture is obtained. An anti-oxidant for living body characterized by having a primary lignin as a main component is disclosed.
JP 2004-238519 A JP 2002-97469 A JP 2002-235084 A Japanese Patent No. 3284097

  As shown in the above Patent Documents 1 to 4, it is known that algae such as Spirulina and Chlorella, and extracts of basidiomycetes such as Agaricus and Meshimakobu have an antioxidant effect.

  However, the above-mentioned conventional antioxidant employs a method in which algae and basidiomycetes are cultured separately, and components extracted from each culture are used alone or in combination. Antioxidants did not have a sufficient effect. Moreover, when using in combination, there existed a problem that the extraction process had to be performed about each raw material, and manufacturing workability | operativity was also bad.

  Accordingly, an object of the present invention is to provide an antioxidant and a food and drink containing the same, which have a higher antioxidant action and can simplify the production process.

  In order to achieve the above object, the antioxidant of the present invention is a culture obtained by culturing mycelia of basidiomycetes and / or ascomycetes on a medium prepared from a raw material containing algae, or the culture It contains the extract prepared from the above as an active ingredient.

  According to the present invention, algae are decomposed and assimilated by basidiomycetes and / or ascomycetes by culturing mycelium of basidiomycetes and / or ascomycetes in a medium prepared from a raw material containing algae. In addition, mycelium of basidiomycetes and / or ascomycetes can be grown. The culture obtained in this way contains a component in which algae are decomposed and assimilated, and a mycelium component of basidiomycetes and / or ascomycetes that have taken up these algae as nutrients. Therefore, this culture or the extract extracted from the culture is a component in which components derived from algae and components derived from mycelium of basidiomycetes and / or ascomycetes are decomposed and fused in a high dimension. It is a new material different from a simple combination of both extracts.

  And, as shown in the examples described later, the culture or its extract is more algae as a raw material, and basidiomycetes and / or ascomycete cultures cultured without using algae as a medium raw material. Compared with, the active oxygen scavenging activity is remarkably high. Therefore, by ingesting the culture or the extract thereof, production of active oxygen in the body can be suppressed and injury due to active oxygen can be prevented.

  Moreover, since algae can be decomposed | disassembled and utilized by the mycelium of a basidiomycete and / or an ascomycete, and these can be extracted together with a culture, a manufacturing process can also be simplified.

  In the antioxidant of the present invention, the algae is preferably chlorella and / or spirulina, and the basidiomycetes and / or ascomycetes are preferably shiitake mushrooms or mannen mushrooms. According to this, a higher antioxidant function enhancing effect can be obtained.

  Moreover, it is preferable to use as an active ingredient the extract obtained by carrying out self-digestion of the said culture with the enzyme contained in the mycelium of the said basidiomycete and / or ascomycete. According to this, mycelia of basidiomycetes and / or ascomycetes themselves are also enzymatically decomposed, so that the active ingredient can be extracted more effectively.

  On the other hand, the food / beverage products of this invention are characterized by containing the said antioxidant. By ingesting this food or drink, production of active oxygen in the body can be suppressed and injury due to active oxygen can be prevented.

  According to the antioxidant of the present invention, it has high active oxygen scavenging activity, and by taking this, it can be expected to suppress the generation of active oxygen in the body and prevent injury due to active oxygen.

  Moreover, since algae can be decomposed | disassembled and utilized by the mycelium of a basidiomycete and / or an ascomycete, and these can be extracted together with a culture, a manufacturing process can also be simplified.

  Furthermore, according to the food / beverage products of this invention, by containing the said antioxidant, an antioxidant can be ingested easily on a daily basis and the disorder | damage | failure by active oxygen can be prevented continuously.

  The basidiomycetes used in the present invention include edible mushrooms such as shiitake mushrooms, Japanese cypress mushrooms, oyster mushrooms, enoki mushrooms, shimeji mushrooms, yamabushi mushrooms, or shochu (truffles); There are various types such as medicinal candy. In addition, examples of ascomycetes include Amiga cocoon. Among these, shiitake mushroom and mannen candy are preferably used.

  Examples of the algae used in the present invention include chlorella, spirulina, euglena, seaweed, kombu, wakame, hijiki, mozuku, human egusa, funori, tosakanori, proboscis, ogonori and giant kelp. Among these, chlorella and spirulina are preferably used. Algae may be used as a raw material as it is, but those subjected to physical treatment such as shredding, crushing and grinding, and chemical treatment such as acid or alkali decomposition and enzymatic decomposition can also be used. Furthermore, the extract extracted with hot water etc. can also be used.

  In the present invention, the mycelia of basidiomycetes and / or ascomycetous fungi are cultured in a medium containing these algae to decompose and assimilate the algae, and the mycelium of basidiomycetes and / or ascomycetous fungi are grown. . In addition to the algae, other nutrient sources necessary for the growth of basidiomycetes and / or ascomycetes may be added to the medium. As the medium, either a liquid medium or a solid medium can be used.

  In the case of a solid medium, a medium containing a vegetable fiber component, for example, a medium in which bagasse (a fiber component of sugarcane), corn stover, rice bran, bran, rice straw, rice bran, bamboo, bear candy, etc. are mixed with algae. Is preferably used. Moreover, you may add and mix yeast extract, dry yeast, okara, corn meal, etc. as needed. The solid medium is adjusted so that the water content is 60 to 80%, sterilized by autoclaving according to a conventional method, then inoculated with mycelium or spores, for example, 1 to 1 in a culture room conditioned at a temperature of 18 to 25 ° C. Incubate for 6 months. In this way, the mycelium spreads inside the medium, and the culture is terminated with the medium pH lowered to 5 or lower. In addition, as shown in the Example mentioned later, in the case of a solid culture medium, since an active oxygen scavenging ability becomes the maximum in 3 months of culture | cultivation immediately before a fruit body generate | occur | produces, culture | cultivation period is 2.5-3. More preferably 5 months.

  In the case of a liquid medium, in addition to algae, carbohydrates (pentose, hexose, their oligosaccharides, hetero-oligosaccharides, polysaccharides, glycoproteins, glycolipids, etc.) that serve as energy sources for basidiomycetes and / or ascomycetes ) Or a medium containing cereals or the like. In liquid culture, raw material algae are finely pulverized as necessary, a carbon source, nutrient components, etc. are added as necessary, and the concentration is adjusted to 1 to 20% with water. Sterilized by autoclaving according to conventional methods, after cooling the medium, inoculated with mycelium germinated and grown from spores, or mycelia separated and cultured from fruit bodies, for example, 7-30 days under temperature controlled at 18-30 ° C. Incubate with aeration or shaking. The culture is completed in a state where the fermentation by the mycelium is sufficiently achieved and the pH of the culture solution is lowered to 5 or less.

  After completion of the culture, the mycelium is self-digested using an enzyme present in the mycelium, the culture medium is crushed, and the culture is extracted.

  In the case of a solid medium, the medium is crushed, a small amount of water is added as needed, and the mixture is treated at 30 to 60 ° C. for 3 to 6 hours to advance the mycelial enzyme reaction and self-digest. Subsequently, this crushed material is infiltrated with warm water or hot water of 50 ° C. or higher to extract active ingredients.

  In the case of a liquid medium, the mycelium is self-digested at 30 to 60 ° C., and then heat treated, for example, at 100 ° C. for 10 minutes to inactivate the enzyme activity and mycelium, thereby separating the water-soluble component and the solid content, respectively. Collect or collect water soluble components and solids together.

In either case of a solid medium or a liquid medium, the extraction can be performed under high pressure, for example, under a vapor pressure of 1 Kg / cm ² and under a high pressure such as 120 ° C.

  The antioxidant of the present invention can be produced as it is by concentrating the extract obtained by the above method as it is or by concentrating it, and pulverizing the extract by freeze drying or spray drying. You can also. When the extract is dried, a fine powder is obtained, which can be further pulverized into ultrafine particles.

  The antioxidant of the product of the present invention thus obtained can be commercialized as a powder, granule, tablet or capsule by a conventional method. Moreover, it can also be commercialized as a liquid and jelly-like drink by adding an extract. Furthermore, the antioxidant of the product of the present invention can be added to various foods and drinks to produce products as foods and drinks that can be expected to enhance the antioxidant function.

  Such foods and drinks are not particularly limited. For example, fresh food such as meat, seafood, vegetables, fruits, etc .; processed livestock products such as ham and sausage; processed marine products such as hampen and kamaboko; jam, dried Processed fruits such as fruits; processed vegetables such as pickles; dairy products such as milk, butter, cream, cheese; oils and fats such as rapeseed oil, palm oil, sunflower oil, shortening; processed soybean foods such as tofu, fried chicken, natto; Beverages such as coffee, cocoa and soft drinks; seasonings such as soy sauce, miso, sauce and ketchup; breads and cakes; confections such as Japanese confectionery and Western confectionery; noodles such as udon, soba noodles, somen and spaghetti.

  The antioxidant according to the present invention contains 30 to 60% protein, 30 to 60% carbohydrate, and 5 to 15% ash, and as shown in the examples described later, eliminates superoxide anion radicals and eliminates nitric oxide. Suppresses production. The effect is remarkably improved as compared with the case where the algae used as a raw material is used alone.

  In the safety test of the antioxidant according to the present invention, no toxicity was shown in a single dose test of 2 g / kg in rats (female), and it was negative in a mutagenicity test with umrack AT. Therefore, it is presumed that the antioxidant of the present invention has extremely low toxicity as expected from a long dietary experience.

  Moreover, the effective dosage of the antioxidant of the present invention is 1 to 10 g per day for an adult when ingested. When the dose is smaller than this, sufficient antioxidative activity in the living body cannot be expected, and when the dose is larger than this, loose stool or abdominal bloating may occur. However, there is no problem in safety even if the dose is larger than the above.

<Example 1> (Manufacture of Spirulina mannentake solid culture fermentation product)
560 ml of water was added to 24 g of Spirulina and 216 g of bagasse (fiber component of sugar cane), mixed well, packed in a polypropylene bag, and sterilized at 121 ° C. for 40 minutes.

  The mycelia of Mannentake mycelium were aseptically added to a liquid medium containing 2% Marz extract and 0.25% yeast extract prepared separately, and cultured with shaking for 20 days.

  Thus, the culture medium inoculated with inoculum of garlic mushroom was allowed to stand at 20 ° C. for fermentation. The culture period was 0 days, 1 month, 2 months, 3 months, 4 months, and 5 months, and the medium was crushed to the size of a thumb and subjected to self-digestion with the enzyme contained in the mycelium of mannentake, at 60 ° C. The extract was obtained in about 15 hours with warm water.

  The extract was roughly filtered through a mesh (12 mesh) and then centrifuged (12,000 rpm, 10 minutes) to obtain a supernatant. Further, the supernatant was concentrated under reduced pressure and then lyophilized to obtain a fine powder.

<Example 2> (Production of chlorella-mannentake solid culture fermented product)
560 ml of water was added to 48 g of chlorella and 192 g of bagasse (a fibrous component of sugar cane) and mixed. The mixed medium material was packed in a polypropylene bag and sterilized at 121 ° C. for 40 minutes. After cooling, 10 ml of Mannentake seeds prepared in the same manner as in Example 1 was inoculated. The mixture was allowed to stand at 20 ° C. for 3 months and 4 months for fermentation. After the fermentation was completed, the medium was crushed to a thumb size and extracted with warm water at 60 ° C. for 15 hours.

  The extract was roughly filtered through a mesh (12 mesh) and then centrifuged (12,000 rpm, 10 minutes) to obtain a supernatant. Subsequently, the supernatant was concentrated under reduced pressure and lyophilized to obtain a fine powder.

<Example 3> (Production of chlorella-mannentake liquid culture fermented product)
A 200 ml Erlenmeyer flask was charged with 5 g of chlorella and 3 g of glucose, 100 ml of purified water was added and dissolved, and then sterilized at 121 ° C. for 20 minutes. The medium was cooled, allowed to reach 23 ° C., and aseptically added 5 ml of liquid culture inoculum of Mannentake cultured in the same manner as in Example 1.

  This was cultured with shaking at 23 ° C. to promote fermentation. Samples were periodically taken and cultured for 14 days, confirming that there was no contamination and that the culture was well. The progress of the culture was judged by measuring the pH of the culture solution.

  At the end of the culture, the mixture was heated at 100 ° C. for 10 minutes to inactivate mycelium and enzymes and freeze-dried to obtain a powdered fermented product.

<Example 4> (Manufacture of Spirulina mannentake liquid culture fermentation product)
Spirulina (1.4 kg), glucose (0.9 kg) and purified water (27 liters) were placed in a 50 L fermenter, sterilized at 121 ° C. for 30 minutes, and the medium was cooled to 23 ° C.

  Inoculate 8L of water into a 10L culture bottle, add 2% malt extract and 0.25% yeast extract, sterilize at 121 ° C for 40 minutes, add mannentake mycelium to the cooled liquid medium, and ventilate sterile air. The culture was continued for 5 days. 3 L of this inoculum was aseptically added and fermented with aeration and stirring at 30 ° C. for 7 days.

  At the end of the fermentation, the mixture was heated at 100 ° C. for 10 minutes to inactivate the mannintake mycelium and the enzyme produced by the mycelium in the culture solution. Subsequently, freeze-dried fine powder of Spirulina mannentake fermented product was obtained by the same operation as in Example 3.

<Example 5> (Production of chlorella shiitake liquid culture fermented product)
A 50 L fermenter was charged with 1.4 kg of chlorella and 0.9 kg of glucose, and 27 L of water was added. After sterilization at 121 ° C. for 30 minutes, the mixture was cooled to 23 ° C. Separately, 3 L of shiitake liquid culture inoculum that was aerated in a 10 L culture bottle was aseptically added. After aeration and agitation culture at 23 ° C. for 14 days, a freeze-dried fine powder of chlorella-shiitake fermentation product was obtained in the same manner as in Example 3.

<Example 6> (Production of Spirulina-shiitake liquid culture fermented product)
A 200 ml Erlenmeyer flask was dissolved by adding 5 g of Spirulina, 3 g of glucose and 100 ml of purified water, and then sterilized at 121 ° C. for 20 minutes.

  Separately, 250 ml of liquid medium (Maltz extract 2%, yeast extract 0.25%) was placed in a 500 ml Sakaguchi flask and sterilized at 121 ° C. for 30 minutes. After cooling, shiitake mycelium was added and shaken for 25 days to produce shiitake inoculum. 5 ml of this inoculum was inoculated after cooling to the sterilized medium. After culturing at 23 ° C. for 14 days, a freeze-dried fine powder of Spirulina-shiitake fermentation product was obtained in the same manner as in Example 3.

<Comparative Example 1> (Raw material Spirulina)
The raw material Spirulina used in Examples 1, 4, and 6 was set as Comparative Example 1. The raw material Spirulina used was Spirulina powder (produced by the People's Republic of China, distributor; Sun Life Co., Ltd.).

<Comparative example 2> (raw material chlorella)
The raw material chlorella used in Examples 2, 3, and 5 was set as Comparative Example 2. The raw material chlorella used was a chlorella bulk powder (produced in Taiwan, distributor; Sun Life Co., Ltd.) produced by producing a seed strain of Pyrenoidosa species by an open culture method and pulverizing it by a spray-drying method.
<Comparative Example 3> (Ganoderma mycelium extract)
The fungus seed bacterium used in Example 4 was centrifuged (17000 rpm, 10 minutes) to collect mycelium. Water was added to the mycelium precipitate, stirred to suspend the mycelium in water, and centrifuged again under the above conditions to remove and wash the culture solution contained in the mycelium. This operation was further repeated twice to wash the washed mycelium.

To the mycelium after lyophilization, warm water at 60 ° C. was added and kept for 15 hours, followed by centrifugation, and the supernatant was lyophilized to obtain a garlic mycelium extract.
<Comparative Example 4> (solid culture fermented product without addition of algae)
216 ml of bagasse and 24 g of rice bran were mixed with 560 ml of water and sterilized at 121 ° C. for 40 minutes. This solid medium was inoculated and cultured with Mannentake seeds in the same manner as in Example 1. Extraction was performed in the same manner as in Example 1 after a culture period of 1 month, 2 months, 3 months, 4 months, and 5 months.

<Test Example 1> (Composition analysis of each sample)
Table 1 shows the result of analyzing the composition of each sample obtained in Examples 1 and 2. In addition, each component analysis was performed by the phenol sulfuric acid method for saccharides, the semi-micro Kjeldahl method for proteins, the acetyl bromide method for water-soluble lignin, and the direct ashing method for ash.

  The results of analyzing the compositions of the samples obtained in Examples 3 to 4 and Comparative Examples 1, 2, and 3 are shown in Table 2 below.

<Test Example 2> (Measurement of superoxide anion radical scavenging activity 1)
For each sample obtained in Example 1, the superoxide anion radical scavenging activity was measured by the following method.

That is, the ability to erase the superoxide anion radical (O ₂ ⁻ ) was measured using chemiluminescence. O ₂ ^- is the generation system using hypoxanthine oxidase Bu system, resulting O ₂ ^- was measured chemiluminescence by reaction with a chemiluminescence reagent in a luminometer. And the luminescence integration value (a) in the state which does not add a test substance and the luminescence integration value (b) at the time of adding a test substance were measured.

  The measurement of the accumulated luminescence value (b) when the test substance is added is as follows. In other words, dispense 10 μl of luminescent reagent (MPEC; ATTO AB-2950) to a 96-well plate and add 10 μl of the test substance. To this, 160 μl of xanthine oxidase and 170 μl of phosphate buffer were added, 50 μl of hypoxanthine solution was added, and the luminescence value was measured at a measurement interval of 1 second from 10 seconds to 110 seconds, and integrated.

  The measurement of the accumulated luminescence value (a) without adding the test substance is performed by adding 10 μl of phosphate buffer instead of the test substance added in the measurement of the accumulated luminescence value (b) when the above test substance is added. The same measurement was performed.

From the above measurement results, the inhibition rate was calculated from the following equation.
Inhibition rate (%) = 1- (b) / (a) x 100
Then, the ratio of the inhibition rate when the inhibition rate of vitamin C is 100 is defined as the relative activity, and the change of the relative activity with the culture period is shown in FIG.

  Moreover, about the solid culture fermented product without the algae of the comparative example 4, the ratio of the inhibition rate when the inhibition rate of vitamin C was set to 100 was set as relative activity similarly to the above, and the change of the relative activity with the culture period was calculated | required. The results are shown in FIG.

  As shown in FIG. 1 and FIG. 2, in Example 1 using spirulina as a medium raw material, the antioxidant activity was remarkably increased when the culture period was 3 months. In contrast, in Comparative Example 4 where spirulina is not included in the medium raw material, it can be seen that the antioxidant activity does not increase so much.

<Test Example 3> (Measurement 2 of Superoxide Anion Radical Scavenging Activity 2)
In the same manner as in Test Example 2, Example 1 (4 months of culture), Example 2 (4 months of culture), Example 3, Example 5, Comparative Example 1 (raw material Spirulina), and Comparative Example 2 The superoxide anion radical scavenging activity was measured for (raw chlorella) and Comparative Example 3 (Ganoderma mycelium extract), and the results are shown in Table 3.

  From the results shown in Table 3, Examples 1, 2, 3, and 5 using spirulina or chlorella as a medium raw material are Comparative Example 1 made of raw material spirulina, Comparative Example 2 made of raw material chlorella, and medium raw materials not containing chlorella or spirulina. It can be seen that the superoxide anion radical scavenging activity is remarkably increased as compared with Comparative Example 3 in which mannentake mycelium was cultured.

<Test Example 4> (Measurement of production amount of nitrogen oxides)
About Example 3, Example 4, Example 5, Example 6, comparative example 1 (raw material spirulina), and comparative example 2 (raw material chlorella), the production amount of nitrogen oxides was measured by the following method.

  Stimulated macrophages produce inducible nitric oxide. Nitric oxide is one of the reactive oxygen species, and chlorella and spirulina give a strong stimulus to macrophages to enhance the production of nitric oxide. Therefore, the influence on nitric oxide production when fermented with basidiomycetes was measured by the following procedure.

  J774.1 cells (available from RIKEN, Cell Bank), a macrophage-like cell line, were suspended in RPMI1640 medium containing 10% FBS and seeded at 100 μl / well on a 96-well plate.

The test substance was appropriately diluted with the above medium and added to the well in which the cells were seeded. Then, after culturing at 37 ° C. under 5% CO ₂ for 3 days, 100 μl of the supernatant was collected in another 96 well plate.

Next, 100 μl of Gries reagent (0.1% naphthylethylenediamide · 2HCl, 2.5% phosphoric acid, 1% sulfanilamide) was added, color was developed for 10 minutes at room temperature, and the absorbance was measured at 540 nm. A calibration curve was prepared using NaNO ₂ as a standard substance.

The amount of nitric oxide produced when LPS (lipopolysaccharide) 2.5 ng / ml was added was taken as 100%, and the concentration producing 50% nitric oxide at each test substance concentration was taken as the ED ₅₀ value. Calculated. The results are shown in Table 4 below.

  As shown in the above Test Examples 1 to 4, the fermented products of Examples 1 to 6 are composed of components of carbohydrates 24 to 63%, proteins 28 to 58%, and ash content 5 to 20%. Compared with Example 1 (raw material Spirulina) and Comparative Example 2 (raw material chlorella), the active oxygen scavenging ability is remarkably increased. In particular, the fermented products of Examples 3 and 5 increased the superoxide anion radical scavenging activity of chlorella before fermentation by 15 times and 38 times, respectively. Moreover, the fermented products of Examples 3 to 6 suppressed the nitric oxide producing ability of Spirulina and Chlorella before fermentation to 1/15 to 1/1149.

  By ingesting the antioxidant according to the present invention and foods and drinks containing the same, the production of active oxygen in the body can be suppressed and injury due to active oxygen can be prevented. Accordingly, it is possible to prevent the occurrence of disorders such as arteriosclerosis, hypertension, myocardial infarction, angina pectoris, cerebral infarction, cerebral hyperemia, diabetes and cancer due to active oxygen. Therefore, it can be widely used as, for example, pharmaceuticals, health foods, and cosmetics.

It is a graph which shows the change of superoxide anion radical scavenging activity by the culture | cultivation period of a Spirulina mannentake solid culture fermented material. It is a graph which shows the change of superoxide anion radical scavenging activity by the culture | cultivation period of the fermented material which culture | cultivated Mannen mycelium in the solid culture medium which does not contain chlorella or spirulina.

Claims (5)

  A medium prepared from a raw material containing algae contains a culture obtained by culturing mycelium of basidiomycetes and / or ascomycetes or an extract prepared from the culture as an active ingredient. Antioxidant.   The antioxidant according to claim 1, wherein the algae is chlorella and / or spirulina.   The antioxidant according to claim 1 or 2, wherein the basidiomycete and / or ascomycete is shiitake mushroom or mannen moth.   The antioxidant according to any one of claims 1 to 4, wherein an extract obtained by self-digesting the culture with an enzyme contained in the mycelium of the basidiomycete and / or ascomycete is an active ingredient. Agent.   Food-drinks containing the antioxidant as described in any one of Claims 1-4.