JP4119656B2 - Antioxidant - Google Patents

Description

[0001]
BACKGROUND OF THE INVENTION
Antioxidant activity effective in preventing quality degradation due to oxidation of food, feed, cosmetics or their raw materials, etc. In vivo oxidation of humans or animals that use it by adding it to food, feed, cosmetics The present invention relates to a natural product-derived antioxidant having an antioxidant activity effective in eliminating or reducing active oxygen and free radicals generated by the above-mentioned method, or reducing the generation of lipid peroxide by them.
The present invention also relates to a food or feed containing an antioxidant derived from natural products. The present invention also eliminates or reduces active oxygen and free radicals generated in the living body of humans or animals using the same, or reduces the generation of lipid peroxides by adding them to foods, feeds and cosmetics. It relates to a therapeutic agent.
[0002]
[Prior art]
In recent years, with the westernization of eating habits, opportunities to ingest high-fat foods have increased, and adult diseases such as hyperlipidemia and arteriosclerosis are increasing. This phenomenon extends not only to middle-aged and elderly people but also to young people. As a result, various diseases are caused by the oxidation of lipids in vivo. Antioxidants are added to foods, feeds and cosmetics and used for the purpose of maintaining the quality, and the antioxidants are administered to the living body, and free of active oxygen generated by in vivo oxidation. Some are used for the purpose of eliminating or reducing radicals and lipid peroxides, that is, preventing or reducing in vivo oxidation.
[0003]
Oxidation of fats, pigments and other active ingredients in foods, feeds or cosmetics during storage causes resinification of the product, generation of off-flavors, coloring, discoloration, production of toxic substances or reduction in nutritional value. For this reason, foods, feeds and cosmetics often contain antioxidants for maintaining quality. Such an antioxidant effect is measured by testing the oxidation inhibition effect and dye fading inhibition effect of fats and oils.
[0004]
For such purposes, antioxidants conventionally used mainly in the fields of food, cosmetics and pharmaceuticals include, for example, natural antioxidants such as tocopherol (vitamin E) and ascorbic acid (vitamin C), and butyl Phenolic synthetic antioxidants such as hydroxyanisole (BHA) and dibutylhydroxytoluene (BHT) are known. Tocopherol is often used as an antioxidant for fats and oils, but its use is limited because it is difficult to dissolve in water, and the essential antioxidant power is not so high, so it has been necessary to use it together with other antioxidants. Ascorbic acid is used as a water-soluble antioxidant and is difficult to use for oils and fats, and when used in foods, its acidity is imparted, changing the taste of the food, and compared with synthetic antioxidants. As a result, its antioxidant power is inferior. On the other hand, synthetic antioxidants such as BHA and BHT have strong antioxidant potential, but their safety is questioned, such as their carcinogenicity and adverse effects on the lungs and liver. became. For these reasons, it has been desired to develop a natural antioxidant having strong antioxidant power and high versatility.
[0005]
Conventionally, the following are reported as natural antioxidants. Antioxidant composition containing water or water-containing alcoholic extract of citrus fruit peel as an active ingredient (Japanese Patent Laid-Open No. 8-259945), antioxidant composition containing extract of citrus kumomikan leaf as active ingredient (Japanese Patent Laid-Open No. 11-199427), foods and drinks containing an antioxidant activity fraction extracted from green leaves of green plants (Japanese Patent Laid-Open No. 11-123071), soluble in water in green leaves of early maturity or n -Antioxidant activity fraction of green leaves of wheat, which is a fraction insoluble in hexane and soluble in water-containing ethanol having a water content of 80% or less and containing 2-O-glucosyl-isovitexin ( Japanese Patent Laid-Open No. 5-65480), an antioxidant extracted from a plant of the genus Phyllidae using an aliphatic alcohol organic solvent having 1 to 5 carbon atoms and / or one or more other organic solvents (Japanese Patent Laid-Open No. 5-156249) Antioxidant substance characterized by extracting and collecting an antioxidant substance contained in a food and cosmetics (Japanese Patent Laid-Open No. 9-59127) and Kufa genus plants with a polar solvent. No. 6,212,154, and an antioxidant containing a hydrolyzate of an olive plant solvent extract as an active ingredient (Japanese Patent Laid-Open No. 9-78061). Antioxidant effects have also been found in extracts of lactic acid bacteria, and the present applicant has disclosed the technology in Japanese Patent Application Laid-Open Nos. H4-264034 and H05-276912. This technique is derived from lactic acid bacteria belonging to Lactobacillus delbruecki and Lactobacillus bulgaricus among lactic acid bacteria. This lactic acid bacterium has no colonization in the human intestinal tract.
[0006]
Conventionally, antioxidants have been regarded as preservatives that prevent oxidation of food, feed, cosmetics and the like. However, in recent years, some antioxidants that have been used in foods so far have an antioxidant effect that eliminates or reduces free radicals, active oxygen, and lipid peroxide generated by in vivo oxidation. Search for natural extracts that have become clear and have strong antioxidant power in vivo (Food & Food Ingredients Journal of Japan, 163, 19-29 (1995), New Food Industry, 39 (3), 17-24 (1997) Monthly Food Chemical 1998-8, P.33-41).
[0007]
Various free radicals and active oxygen generated by in vivo oxidation are known. Since free radicals are a general term for molecular species having unpaired electrons, there are various in vivo free radicals. Superoxide anions and hydroxyl radicals are known as free radicals that are also active oxygen. Hydrogen peroxide and singlet oxygen are known as active oxygens that are not free radicals. Among these, those generally measured as indicators of in vivo antioxidant effects are radical scavenging activity using 1,1-diphenyl-2-picrylhydrazyl (DPPH), superoxide anion scavenging activity, hydroxyl radical These are scavenging activity and lipid peroxide production inhibitory activity, and those having these activities are generally regarded as antioxidants having in vivo antioxidant power (Monthly Food Chemical 1998-8, P.33-41).
[0008]
Recently, methods for measuring oxidative damage at the genetic level have been studied in assessing the potential of food aging prevention at the clinical level. Among them, 8-hydroxydeoxyguanosine (8-OHdG) was generated as a result of the attack on nucleobase (deoxyguanosine) in DNA by active oxygen resulting from lipid peroxidation in vivo. It was found that 8-OhdG is usually excised by DNA repair enzyme, and finally excreted in blood after passing through blood. Therefore, the anti-aging effect can be measured by measuring the amount of 8-OHdG in blood or urine, but this method uses the ELISA method, which is an immunochemical method, and this method is still expensive. Is not established as a general method. Therefore, at present, the above-mentioned free radical scavenging activity and active oxygen scavenging activity are usually measured as indicators of the in vivo antioxidant effect.
[0009]
Currently, the following are known as diseases considered to be caused by in vivo oxidation. Brain and nervous system diseases include cerebral edema, traumatic epilepsy, cerebral ischemia, Parkinson's disease, spinal cord injury, etc. Eye diseases include cataract, retinal degeneration, retinopathy of prematurity, etc. as respiratory diseases Bronchial asthma, emphysema, pulmonary fibrosis, adult respiratory distress syndrome (ARDS), premature infant respiratory distress syndrome (IRDS), etc., and cardiovascular diseases include arteriosclerosis, myocardial infarction, etc. Ulcerative colitis, stress gastric ulcer, pancreatitis, gastrointestinal mucosal disorder, etc., skin diseases include ultraviolet rays, atopic dermatitis, burns, frostbite, bed sores, etc., kidney diseases include nephritis, kidney Other diseases such as insufficiency include cancer, diabetes, collagen disease, rheumatism, Alzheimer's disease, autoimmune disease, Behcet's disease and the like. Aging, skin wrinkles and dementia due to aging are also considered to be due to in vivo oxidation. Antioxidants that prevent or reduce in-vivo oxidation are thought to lead to the prevention and treatment of such diseases, and have attracted attention as functional foods or health foods.
[0010]
Antioxidants for such applications include active oxygen scavengers (JP-A-8-20544) containing as an active ingredient an extract obtained by extracting hantaikai with water or a water-containing organic solvent, plant seeds or germs Anti-oxidative oxygen action composition characterized in that after roasting potatoes, the composition obtained by adding plant seeds to the enzyme-treated one is washed with a nonpolar solvent, and then insoluble matter is extracted with a polar solvent. Product (JP-A-4-139132), active oxygen scavenger (JP-A-6-65074) characterized in that it contains astilbine extracted from oysters, and yellowfin fruit is extracted with a solvent such as water or methanol An anti-oxidant extract (Japanese Patent Laid-Open No. 10-17485), an essence of a novel mint hybrid, an active oxygen scavenger (Japanese Patent Laid-Open No. 10-66465), and a medium containing a plant fiber component Basidiomycetes An antibacterial function-enhancing agent for living organisms comprising as a main component a saccharide, protein, and water-soluble lignin obtained by culturing mycelium of ascomycetes and extracting from the culture No. 228441), active oxygen scavenger characterized by containing one or more plant extracts selected from Akaminoki, Uyaku, Uwaurushi, Zicopi, Perilla, Tencha, Lotus, and Lavender (Japanese Patent Laid-Open No. 11-279069) JP, No. 10-195433), superoxide dismutase-like activity derived from solvent extraction of proliferative cell cultures derived from sesame plants Substances (JP-A-4-275226) have been reported. In addition, rooibos tea extract, dried ginkgo biloba extract, etc. are on the market.
[0011]
In addition, as an antioxidant that has both an antioxidant effect for maintaining quality and an in vivo antioxidant effect, the active ingredient is a substance obtained by extracting with a polar solvent from leaves and stems of fennel, which is a kind of spice plant An antioxidant (Japanese Patent Laid-Open No. 6-299151), an antioxidant (Japanese Patent Laid-Open No. 10-46142) characterized in that it comprises a plant extract of the family Chironomidae as an active ingredient are disclosed. . It is also known that tea extract, grape seed extract and the like suppress in vivo oxidation.
However, it is not known that lactic acid bacteria showing human intestinal colonization suppress the in vivo oxidation.
[0012]
[Problems to be solved by the invention]
Conventionally reported antioxidants for maintaining quality include synthetic products and natural extracts. Synthetic products have safety problems, so natural and highly safe antioxidants are required. However, natural antioxidants also have various problems and are required to be improved.
[0013]
For example, antioxidants that are basically fat-soluble, such as tocopherol, sesame extract, and sunflower seed extract, are difficult to use as they are in water-based products, and water-soluble antioxidants are oils and fats. There was a drawback that it was difficult to use. Moreover, since the antioxidant which has a peculiar taste and flavor influences the taste and flavor of a product, the use range was limited. Therefore, a natural antioxidant that has an antioxidant effect in a small dose within the range where the taste and odor imparted to a wide range of foods, feeds and cosmetics is acceptable is desired.
[0014]
There are also various problems with in vivo antioxidants known so far. Conventionally, plant extracts such as those known as Kampo, herbs, or folk medicines have medicinal effects other than antioxidant effects, and side effects due to the medicinal effects sometimes occur. For these reasons, safe plant-derived natural antioxidants that have been conventionally treated as foods are desired.
In addition, conventional natural antioxidants are cultivated and used for the purpose of extracting antioxidants, and thus have a problem of high cost.
[0015]
[Means for Solving the Problems]
In view of the above problems, the present inventors have intensively studied a versatile antioxidant that is safe for humans or animals and can be produced at low cost.
The present inventors have been researching various fermented milks, and among the lactic acid bacteria isolated from these fermented milks and lactic acid bacteria derived from humans, the high intestinal intestinal level that has not been conventionally found in Lactobacillus gasseri. Furthermore, the present inventors have found that the Lactobacillus gasseri lactic acid bacterium and its mutant strain culture and microbial cells have an in vivo antioxidant effect and succeeded in solving the above problems.
[0016]
  In this specification, the “antioxidant effect” means that when a food or feed is administered to humans or animals, free radicals and active oxygen generated in the living body are erased or reduced to exert a cell membrane stabilizing effect. Including in vivo antioxidant effect. An “antioxidant” is an agent having such an antioxidant effect. That is, the present invention relates to a microbial cell obtained by culturing lactic acid bacteria belonging to Lactobacillus gasseri, and an antioxidant containing the culture as an active ingredient. Furthermore, the present invention has an antioxidant action comprising such an active ingredient.feedAbout. That is, the present invention is an invention having the following configurations described in the claims.
[0017]
(1) Lactobacillus gasseri having intestinal colonization ・ SBT2055 ( FERM P-15535 )As an active ingredient, the culture and / or the bacterial cells obtained by culturingHas in vivo antioxidant effectAntioxidants.
(2) Lactobacillus gasseri ・ SBT2055 ( FERM P-15535 )The culture obtained by culturing is fermented milk,Has in vivo antioxidant effectAntioxidants.
(3)Lactobacillus gasseri ・ SBT2055 ( FERM P-15535 )Added culture and / or cells obtained by culturingIn vivo antioxidant effectFeed.
[0018]
DETAILED DESCRIPTION OF THE INVENTION
The present invention has been made in order to solve the above-mentioned problems, and the present inventors newly set the following criteria and screened strains that meet the purpose when screening the target lactic acid bacteria. Selected. That is, the present inventors, among many lactobacillus gasseri derived from fermented milk and humans, have high gastric acid resistance, good growth under low pH conditions, suppress the increase in serum cholesterol, human intestinal tract High fixability, high affinity for human intestinal cells, bile acid resistance, strong inhibition of in vivo oxidation, high survival when applied to foods, excellent flavor, physical properties, etc. Conditions were set, and intensive research was conducted on the selection of strains. No Lactobacillus gasseri bacteria satisfying such conditions have been known. As a result of screening under these conditions, the following strains could be selected as strains meeting these conditions. These strains are deposited at the Patent Organism Depositary, National Institute of Advanced Industrial Science and Technology under the following deposit number.
[0019]
Strain
Lactobacillus gasseri SBT2055 FERM P-15535
furtherthisThe strain has a high affinity for human intestinal cells, and when orally administered, it can survive and reach the intestinal tract and can be resident in the intestinal tract for a long period of time. And it is not known at all that lactic acid bacteria belonging to Lactobacillus gasseri colonize in the intestine and prevent in vivo oxidation, and were revealed for the first time by the present inventors.It was.
[0020]
Next, the culture method of these lactic acid bacteria is described.
Lactobacillus gasseri (Lactobacillus gasseriFor the culture medium of), various media such as a milk medium or a medium containing milk components, and a semi-synthetic medium not containing this can be used. An example of such a medium is a reduced skim milk medium obtained by reducing skim milk and then heat sterilizing.
The culture method is a static culture or a neutralization culture with a constant pH controlled. However, the culture method is not particularly limited as long as the bacteria grow well.
[0021]
In the present invention, the culture and / or cells obtained as described above are used as active ingredients. A dried powder may be used as the active ingredient. Such drying is preferably performed by freeze-drying because the cells can be dried without deteriorating the cells.
These active ingredients are preferably taken orally. When ingesting, it may be added to a high-cholesterol-containing food and taken at the same time, or it may be taken before and after the intake of a high-cholesterol food. These powders can be mixed with an appropriate excipient such as lactose and orally administered as a powder, tablet, pill, capsule or granule. The dosage is appropriately determined individually in consideration of the symptom, age, etc. of the subject of administration, but is usually 0.5 to 50 g as a dry product per day for an adult, and this may be divided into several times a day. In the present invention, a stronger effect is exhibited when Lactobacillus gasseri colonizes in the human intestinal tract. For this reason, it is particularly preferable that 10 per adult as a viable bacterium.⁸~Ten¹²By administering cuf / day, the intended effect of the present invention can be exhibited. By taking in this way, it settles in the intestinal tract and exhibits the desired effect.
[0022]
Moreover, you may add the active ingredient of this invention to a raw material during the manufacturing process of food-drinks. The food and drink may be any food and drink, for example, beverages such as fruit juice, milk drinks and soft drinks, confectionery, dairy products such as butter with a high lipid content, processed egg products such as mayonnaise, butter cake Foods such as However, as a characteristic of the present invention, it is necessary to settle in the intestinal tract while the lactic acid bacteria are alive, and excessive heating must be avoided. It would also be necessary to take measures to avoid heating by employing conventional techniques such as microcapsules.
Furthermore, the food and drink in the present invention may be yogurt or the like produced by lactic acid fermentation using the aforementioned strain of Lactobacillus gasseri.
[0023]
Test examples using LG2055 (FERM P-15535) as the lactic acid strain used in the present invention will be shown below, and the mycological properties and in vitro and in vivo effects will be specifically described. However, the present invention is not limited to this description.
Properties of lactic acid bacteria
[0024]
[Table 1]
[0025]
Genetic characteristics
DNA homology test
The Lactobacillus standard strain described below, the test strain LG2055 strain, and Escherichia coli DNA as controls were extracted and purified.
Test bacteria: LG2055 strain
Reference strain: Lactobacillus acidophilus JCM1132 strain
Lactobacillus crispatas JCM1185 strain
Lactobacillus gallinarum JCM2011 strain
Lactobacillus amylobolus JCM1126 strain
Lactobacillus gasseri JCM1131 strain
Lactobacillus johnsonii JCM2012 shares
Escherichia coli
DNA homology between the LG2055 strain and each of the reference strains was examined by DNA hybridization, assuming that the homology between the DNA of the LG2055 strain was 100% and the DNA homology between the LG2055 strain and E. coli was 0%. As a result, since the LG2055 strain had 90% or more homology with the Lactobacillus gasseri JCM1131 strain, the LG2055 strain was identified as Lactobacillus gasseri.
[0026]
Stomach acid tolerance
In the gastric acid tolerance test, an artificial gastric fluid having a pH of 2.0 was prepared according to the method of Higuchi et al. (Intestinal Bacteriology Journal 14.11-18.2000), and the gastric acid tolerance test was conducted. The LG2055 strain survived for 3 hours or more.
[0027]
Artificial intestinal fluid resistance
An artificial intestinal fluid containing bile powder was prepared according to the method of Higuchi et al. (Intestinal Bacteriology Journal 14.11-18.2000), and the above-mentioned LG2055 strain treated with artificial gastric juice was added to this and tested for resistance. The above viability was demonstrated. The LG2055 strain was confirmed to pass through the gastrointestinal tract and survive to reach the large intestine.
[0028]
In vitro antioxidant activity
B. Halliwell et al.'S deoxyribose method (Analytical Biochemistry, Vol.165, pp.215-219 (1987)) showed a hydroxyl radical scavenging effect when examined using the culture product of strain LG2055.
Moreover, SOD like activity was shown when SOD like activity was measured using SOD Test Wako (made by Wako Pure Chemical Industries Ltd.).
[0029]
Human intestinal transit ability and intestinal colonizationsex
  NothingFermented milk produced by inoculating 4% of LG2055 starter into milk with 9.5% solid milk fat and 3.0% milk fat and fermented at 39 degrees for 4 hours for 42 healthy adult volunteers once a day, 100g daily for 4 weeks The change of enteric bacteria was observed by feeding. During the test period, evaluation was performed by eating freely except forbidding foods, oligosaccharides, and drugs that affect enterobacteria. The LG2055 strain, which was not detected before the test, was detected in all subjects after 4 weeks, indicating that the LG2055 strain has high intestinal colonization.
[0030]
In vivo antioxidant effect test in vivo
1. Preparation of lactic acid bacteria skim milk culture
Using LG2055, cultivated at 37 ° C. for 16 hours in an 11.55% skim milk medium supplemented with 0.5% yeast extract (manufactured by Asahi Breweries) sterilized at 115 ° C. for 20 minutes and activated for 3 or more generations. This was inoculated with 3% of the same medium and cultured at 37 ° C. for 16 hours. The obtained culture was freeze-dried and then ground in a mortar.
[0031]
2. Test feed
Diet composition
Table 2 shows the diet composition based on AIN-76TM (manufactured by Oriental Yeast). 10% non-fat dry milk or the above non-fat dry milk lactobacillus culture was added to this feed, and further, casein was added in consideration of the amount of protein contained in the additive.
[0032]
[Table 2]
[0033]
  The crude protein content (%) of skim milk powder culture and skim milk powder was 35.0 and 34.1, and there was no difference in amino acid composition. In order to facilitate the evaluation of the effects of antioxidant components, a feed containing vitamin E lower than the normal vitamin E level in rodent feed was prepared. For this purpose, a vitamin E-free vitamin mixture was used. In addition, commercially available safflower oil (9.4 mg α-tocopherol) was used as a fat and oil source (feeds containing the respective fats and oils were used as high vitamin E and low vitamin E feeds). Vitamin E was not detected in both dairy products. The skim milk powder culture contained higher concentrations of succinic acid, lactic acid, formic acid, and butyric acid as compared to skim milk. Skim milk and safflower oil or activated carbon treatmentReasonFeed groups containing flower oil were classified into groups 1 and 3, respectively. Non-fat dry milk culture and safflower oil or activated carbon treatmentReasonFeed groups containing flower oil were divided into 2 and 4 groups, respectively.
[0034]
Experimental animals
Five week old Sprague-Dawley male rats (12 rats per group) were used. The animals were reared for 4 weeks with the feed shown in Table 2. During this time, water was given freely. After breeding, the animals were killed by aortic blood sampling under ether anesthesia.
[0035]
analysis
According to conventional methods, hemolytic reaction inhibitory activity of erythrocytes, serum, thiobarbituric acid reactive substance (TBARS) concentration of liver and erythrocytes, α-tocopherol concentration of plasma and liver, and glutathione concentration of liver were measured. For plasma and liver homogenates (0.25 ml), FeSO4 and cumene hydoroperoxide were added and incubated for a certain period of time, and peroxide production was measured using TBARS as an index.
[0036]
Statistical processing
Results were expressed as mean values and standard errors, and Duncan's multiple comparison test and ANOVA test were performed.
[0037]
Experimental result
As is apparent from Table 3, the weight gain and food intake of the Sprague-Dawley rats were not affected by the diet, but the liver weight was slightly reduced in the high vitamin E diet-fat milk group.
[0038]
[Table 3]
[0039]
The effect of diet on erythrocyte hemolysis inhibitory activity is shown in FIG. In the low-vitamin E-fat milk group (3), the erythrocyte hemolysis inhibitory activity was slightly reduced. On the other hand, in the low vitamin E-fermented skim milk group, the hemolysis inhibitory activity was not reduced.
[0040]
The TBARS concentration of erythrocytes was lower in the low vitamin E diet rats (3,4) than in the high vitamin E diet rats (1, 2) (FIG. 2). Similar to the case of erythrocytes, the TBARS concentration of plasma not treated with cumene hydoroperoxide was also decreased in the low vitamin E diet rats (3,4) (FIG. 3). In this case, the TBARS concentration tends to be lower in the fermented skim milk group (2,4) than in the corresponding skim milk group (1,3). In particular, the TBARS concentration in the high vitamin E-fermented skim milk group is There was a significant decrease compared to the corresponding skim milk group (FIG. 3).
Plasma TBARS concentration increased with hydroperoxide treatment, but the increase was significant in rats with low vitamin E diet (3,4), and the increase was observed in the fermented skim milk group (4) and the skim milk added group (3) More suppressed.
[0041]
FIG. 4 shows the TBARS concentration in the liver. Unlike the case of red blood cells and plasma, the TBARS concentration of the liver homogenate not treated with hydroperoxide was lower in the low vitamin E diet rats (3,4) than in the high vitamin E diet rats (1, 2). Hydroperoxide treatment increased the TBARS concentration of liver homogenate, particularly in rats with low vitamin E diet (3,4). In this case, the influence of fermented skim milk addition was not seen.
[0042]
FIG. 5 shows the glutathione concentration in the liver. The reduced glutathione concentration was higher in the low vitamin E diet rats (3, 4) than in the high vitamin E diet rats (1, 2). There was no effect of fermented skim milk addition on the reduced glutathione concentration. Oxidized glutathione concentration was higher in the high vitamin E-fat milk group (1) than in the corresponding fermented skim milk group (2) and other groups (3,4).
[0043]
Plasma, red blood cell, and liver α-tocopherol concentrations are shown in FIG. As expected, these α-tocopherol concentrations were significantly lower in the low vitamin E diet rats (3,4) compared to the high vitamin E diet rats (1,2). The α-tocopherol concentration in plasma tends to be higher in the fermented skim milk group (2,4) than in the corresponding skim milk group (1,3), in particular, the low vitamin E rat feed-fermented skim milk group (4) Compared with the corresponding skim milk group (3), it was significantly higher. The effect of adding fermented skim milk was not observed on the erythrocyte α-tocopherol concentration. In the liver, the α-tocopherol concentration was higher in the high vitamin E-fermented skim milk group (2) than in the corresponding skim milk group (1).
[0044]
Table 4 shows plasma and liver lipid concentrations. Serum triglyceride concentration is higher in low vitamin E diet rats (3,4) than in high vitamin E diet rats (1,2), and in skim milk group (1,3) in fermented skim milk group (2,4) ). There were no dietary effects on plasma cholesterol and phospholipids or liver triglyceride levels. Liver cholesterol and phospholipid concentrations were lower in rats with low vitamin E diet (3,4) than in rats with high vitamin E diet (1,2).
[0045]
[Table 4]
[0046]
As a result, the following was found.
In this experiment, the condition in which the oxidative stress in the living body is different was set by adjusting the vitamin E level of the rat diet. To that end, vitamin E was removed from the dietary vitamin mix. Moreover, since vitamin E is contained in commercially available vegetable oil, vitamin E was removed by treating the vegetable oil used for feed with activated carbon. And the level of vitamin E in the feed is about 98.8% (high vitamin E diet) and 18.8% (low vitamin E diet) of the amount supplied from the standard diet prepared using the vitamin mixture of AIN-76 diet The two stages were set. As a result, when a low vitamin diet was given, the fermented milk of the present invention exhibited an antioxidant action against the enhancement of the peroxidation reaction by plasma cumene hydoroperoxide. In addition, the fermented milk of the present invention was also effective for erythrocyte hemolysis inhibitory activity. The decrease in plasma vitamin E levels in the low vitamin E diet rats was higher in the skim milk group than in the fermented skim milk group of the present invention.
[0047]
Therefore, it can be judged that the fermented skim milk of this invention acted on the increase in the peroxide accompanying the increase in the oxidative stress in the living body and the decrease in the hemolysis inhibitory activity through suppression of vitamin E consumption.
As a result of the above in vivo experiments, it was confirmed that the present invention has an in vivo antioxidant effect.
[0048]
The present invention will be specifically described below with reference to examples. However, the present invention is not limited to this embodiment.
【Example】
Example 1
The LG2055 strain was cultured in a 10% reduced skim milk medium (121 ° C., heated for 10 minutes), and the main culture was freeze-dried and powdered to prepare an antioxidant (A).
[0049]
Example 2
The LG2055 strain was inoculated into a yogurt mix (2% skim milk was added to raw milk and heated at 100 ° C. for 10 minutes) and cultured at 20 ° C. for 24 hours. After filling into a paper cup and cooling, antioxidant yogurt was obtained (B).
[0050]
Example 3
The LG2055 strain was cultured in whey medium (0.5% yeast extract and 0.1% trypticase peptone added), and the cells were collected by centrifugation. 1 g of this microbial cell was mixed with 5 g of lactose and formed into granules to obtain granules.
[0051]
Example 4
The example mix | blended with the foodstuff containing high cholesterol is shown.
Fermented butter (wt / wt)
Milk fat 96.8%
Salt 1.2
Antioxidant (A) 2 obtained in Example 1
[0052]
Butter cake (wt / wt)
Butter 24%
Light flour 24
Sugar 24
Whole egg 24
Antioxidant (B) 4 obtained in Example 2
Fragrance a little
[0053]
Mayonnaise (wt / wt)
Salad oil 65.4%
Egg yolk 17
Vinegar 10
Antioxidant (A) 3 obtained in Example 1
Spice 4.4
Monosodium glutamate 0.6
[0054]
Example 5
The LG2055 strain was inoculated into 5 L of MRS liquid medium (Difco), followed by static culture at 37 ° C. for 18 hours. After completion of the culture, centrifugation was performed at 7,000 rpm for 15 minutes to obtain 1/50 amount of concentrated bacterial cells of the culture solution. Next, the same amount of the concentrated cells was mixed with a dispersion medium containing 10% (weight) of skim milk powder and 1% (weight) of sodium glutamate, adjusted to pH 7, and then lyophilized. The resulting lyophilized product was pulverized with a 60 mesh sieve to obtain a lyophilized bacterial powder.
[0055]
Example 6
In accordance with the provisions of the 13th revised Japanese Pharmacopoeia General Rules for Preparations, “Powder”, 1 g of freeze-dried bacterial strain of LG2055 obtained in the above example was added 400 g of lactose (JP) and 600 g of potato starch (JP). In addition, it was mixed uniformly to produce a powder.
[0056]
Example 7
The skim milk was sterilized at 80 to 85 ° C. for 20 to 30 minutes, and then homogenized and cooled. 2-5% of a pure culture of this strain LG2055 was added to this as a starter and fermented at 37-40 ° C. for 16 hours to obtain acid milk (culture in skim milk medium) having a lactic acid content of 2%. Next, the resulting curd was crushed and cooled to 5 ° C., and this was used as acid milk. Separately, in addition to 15% sucrose, prepare a sugar solution containing appropriate amounts of acidulant, flavor, and pigment, homogenize, sterilize at 70-80 ° C for 20-30 minutes, cool to 5 ° C, did. A sour milk beverage was obtained by mixing the sour milk 35 thus obtained at a ratio of the sugar solution 65.
[0057]
Example 8
40 g of vitamin C or 40 g of an equal mixture of vitamin C and citric acid, 100 g of granulated sugar, 60 g of an equal mixture of corn starch and lactose, 40 g of the freeze-dried product of the culture in the skim milk medium of this strain obtained in Example 1 above In addition, well mixed. The mixture was packed in a bag to produce 150 bags of 1.5 g of sticky nutritional health food.
[0058]
Example 9
An in vivo antioxidant was prepared by the following formulation.
(1) 50 g freeze-dried culture of LG2055 strain nonfat dry milk medium,
(2) 90 g lactose,
(3) Corn starch 29g,
(4) 1 g of magnesium stearate and this mixture were compressed by a compression tablet machine to produce 100 tablets containing 40 mg of active ingredient per tablet.
[0059]
【The invention's effect】
According to the present invention, Lactobacillus gasseri (Lactobacillus gasseriAs an antioxidant containing as an active ingredient the microbial cells obtained by culturing lactic acid bacteria belonging to), an antioxidant containing lactic acid bacteria and / or a mutant thereof as a main component can be provided. The antioxidant of the present invention has extremely low toxicity and side effects, and is useful as a food material.
[Brief description of the drawings]
FIG. 1 shows erythrocyte hemolysis inhibitory activity of domestic rats.
FIG. 2 shows the amount of erythrocyte thiobarbituric acid reactive substance in domestic rats.
FIG. 3 shows the amount of thiobarbituric acid reactive substance in plasma.
FIG. 4 shows the amount of thiobarbituric acid reactive substance in the liver.
FIG. 5 shows glutathione concentration in the liver.
FIG. 6 shows vitamin E concentrations in plasma, red blood cells, and liver.

Claims (3)

Antioxidant with Lactobacillus gasseri (Lactobacillus gasseri) · SBT2055 (FERM P-15535) in vivo antioxidant effect of the cultures and / or cell of the active ingredient obtained by culturing with human gut fixability Agent. Lactobacillus gasseri · SBT2055 culture obtained by culturing (FERM P-15535) is fermented milk, an antioxidant having in vivo antioxidant effect of claim 1, wherein. Lactobacillus gasseri · SBT2055 (FERM P-15535) culture obtained by culturing and / or fodder with the added vivo antioxidant effect the cells.