KR20130092182A - Lactobacillus brevis with high alcohol dehydrogenase activity and dairy products, health functional food and food additives comprising the same - Google Patents

Abstract

PURPOSE: Lactobacillus brevis HD-01 is provided to effectively break down alcohol with high alcohol dehydrogenase activity, to reduce hangover with high acetaldehyde dehydrogenase activity, and to have excellent resistance to acid and bile. CONSTITUTION: Lactobacillus brevis HD-01 (deposit number KACC91701P) has a high alcohol dehydrogenase activity and a high acetaldehyde dehydrogenase activity. A daily product contains the strain. A health food for preventing alcoholic liver disease or reducing hangover contains the strain. A food additive contains the strain. [Reference numerals] (AA) Blood alcohol concentration(mg/dl); (BB) Elapsed time after drinking alcohol(hr); (CC) Comparative example; (DD) Example

Description

Lactobacillus brevis with high alcohol dehydrogenase activity and dairy products, health functional food and food additives comprising the same}

The present invention relates to Lactobacillus brevis excellent in alcohol resolution and dairy products, nutraceuticals and food additives containing the same.

The liver manages energy metabolism in our body as a whole and has the ability to store or distribute various metabolites and compounds such as bilirubin, bile acids, cholesterol, and phospholipids throughout the body, and to detoxify or serve as a major immune organ. It is an organ.

Currently, the cause of mortality caused by disease in the country, the rank of death from liver disease is ranked 4th with 22.9 people per 100,000 population (as of 2001), and the liver disease due to alcohol consumption can not be ignored. According to the World Health Organization's alcohol consumption survey in 153 countries in 1996, Korea's adult alcohol consumption per capita was 14,4ℓ per year, ranking 2nd in the world, and 23,000 people died of accidents and diseases caused by alcohol every year. It also announced that the economic loss amounted to 16 trillion won. In addition, the proportion of alcoholics among Korean adults is 21.6%, the highest in the world, and one out of five adults is suffering from severe national illness as an alcoholic.

Alcoholic liver disease is acute / chronic liver disease with hepatocellular damage caused by persistent and excessive drinking. In the liver, there are enzymes that break down alcohol and decompose alcohol through aldehydes. Acetaldehyde is toxic to liver cells. It will be damaged.

These alcoholic liver diseases are classified into alcoholic fatty liver, alcoholic hepatitis, and alcoholic cirrhosis. The mechanism of liver damage may be due to alcohol itself, metabolites such as acetaldehyde, or immune response. Peroxidation of fat, binding to cytoplasm, disruption of electron transport systems in mitochondria, disruption of microtuble, formation of proteins and binding substances, increased collagen synthesis, etc.

Alcohol is absorbed by about 20% in the stomach and most of the rest is absorbed in the small intestine, 80-90% of the metabolism in the liver, so much of the liver's alcohol metabolism can be treated in the small and large intestine, less burden on the liver, further It may be able to suppress liver damage.

Therefore, because alcohol is not stored in the body, it must be metabolized, which is mostly done in the liver, and this metabolism is done by alcohol-degrading enzymes present in the liver. Alcohol is broken down through acetaldehyde by these enzymes, which are toxic and damage liver cells. And, as a result of alcohol metabolism, many fatty acids are made and fat is accumulated in the liver, which is called alcoholic fatty liver. The alcoholic fatty liver is often progressed to chronic liver disease, alcoholic hepatitis is said to occur in 10 to 35% cirrhosis in 8 to 20%.

Recently, probiotics, which have been in the spotlight as enteric beneficial bacteria, have been widely applied to the prevention and treatment of diseases. In particular, it is widely known that they have the effect of inhibiting proliferation and decay activity against intestinal rot and pathogens (Schrezenmeir J, De Vrese M: Am J Clin Nutr. 73 (2001), P 361S-4S). Probiotics have been reported to reduce the amount of toxic substances by inhibiting the growth of harmful bacteria that produce ammonia and amines, thereby reducing the amount of ammonia and alpha amino nitrogen in the blood and improving symptoms of liver disease. In addition, it has been reported to protect the liver from alcohol and improve the specific function of the liver among these useful bacteria (Raibaud, P: The 3rd International Synposium on Lactic Acid Bacteria and Human Health. (1983), P116-126). Recently, useful lactic acid bacteria such as Probiotics lactobacillus and Bifidobacterium actively promote alcohol metabolism to rapidly break down alcohol and metabolize acetaldehyde harmful to human A study of the possibility of detoxification has been reported (Nosava. T. et al: Alcohol and Alcoholism. 35 (2000), P561-568).

Korean Patent No. 0543115 discloses a strain of Lactobacillus brevis HY7401, which is isolated from feces of Korean adults and which is excellent in ethanol and acetaldehyde degrading enzyme activity and reducing ability, and has been filed and registered by the present applicant. 0769299 discloses Lactobacillus permanent KCTC 10713BP, which is resistant to alcohol and possesses an alcohol degrading enzyme and thus the ability to degrade alcohol.

However, these strains did not have sufficient alcohol dehydrogenase activity to achieve alcohol degradability suitable for the manufacture of alcohol-degradable dairy products or health foods, and also with alcohol dehydrogenase activity as hangover and acute hepatotoxicity. There was a limit in which the activity of acetaldehyde dehydrogenase, which is a cause of, was not high.

The present invention has a high alcohol dehydrogenase activity and excellent alcohol decomposing ability, and at the same time, acetaldehyde dehydrogenase activity has high hangover relieving activity, excellent acid resistance and bile resistance, and gas forming is weak in dairy products. It is an object to provide Lactobacillus brevis with little impact on quality and taste during storage.

The present invention provides Lactobacillus brevis HD-01 [Accession Number: KACC91701P] excellent in alcohol resolution.

Lactobacillus brevis of the present invention brevis ) HD-01 [Accession No .: KACC91701P] is characterized by superior alcohol dehydrogenase activity and acetaldehyde dehydrogenase activity at the same time.

The present invention provides a dairy product containing Lactobacillus brevis HD-01 [Accession No. KACC91701P] excellent in alcohol resolution.

The dairy product of the present invention is characterized in that the fermented milk, butter, cheese or milk powder.

The present invention provides a dietary supplement for preventing or hangover of alcoholic liver disease containing Lactobacillus brevis HD-01 [Accession Number: KACC91701P] excellent in alcohol resolution.

Hangover health functional food of the present invention is characterized in that the powder, granules, tablets or capsule form.

The present invention provides a food additive containing Lactobacillus brevis HD-01 [Accession Number: KACC91701P] excellent in alcohol resolution.

Lactobacillus brevis of the present invention brevis ) HD-01 [Accession No .: KACC91701P] has high alcohol dehydrogenase activity and excellent alcohol decomposability, and at the same time, acetaldehyde dehydrogenase has high activity to relieve hangover, and is resistant to acid and bile. Since it is excellent, it can be used as a dairy product, dietary supplement or food additive to prevent alcoholic liver disease or to relieve hangover.

1 is a graph showing the degradation effect after the alcohol intake of the group ingested Lactobacillus brevis HD-01 of the present invention in Experimental Example 6, it is a graph showing the change in blood alcohol concentration over time.

The lactobacillus brevis HD-01 of the present invention is separated from kimchi and has high alcohol dehydrogenase activity, and thus has high alcohol decomposability, and at the same time, high acetaldehyde dehydrogenase activity, which is excellent in hangover resolution, acid resistance and bile resistance. Excellent in sex

In addition, when the Lactobacillus brevis HD-01 of the present invention is cultured in skim milk, the proliferation of lactic acid bacteria is active, and when it is added to dairy products, especially fermented milk, because it does not affect pH, acidity and curd production without generating gas. Functionality such as alcohol degradability and hangover ability is imparted but little effect on product properties or shelf life.

Lactobacillus brevis HD-01 of the present invention can be used as an active ingredient in dairy products, health functional foods, food additives for preventing or hangover alcoholic liver disease.

The Lactobacillus brevis HD-01 of the present invention may include live bacteria, crushed cell wall fractions, dead bacteria, dried bacteria or cultures as an active ingredient, and may further include an excipient or a carrier. The culture includes a culture solution itself cultured in a liquid medium, a filtrate (centrifuged supernatant) from which the strain is removed by filtration or centrifugation of the culture solution.

The dairy product according to the present invention is fermented milk, butter, cheese or milk powder containing Lactobacillus brevis HD-01, and the health functional food is in the form of powder, granules, tablets or capsules containing Lactobacillus brevis HD-01, and a food additive Is a variety of foods, for example, beverages, sweets, diet bars, chocolate, pizza, ramen, other noodles, gums, ice cream and the like.

In order to prepare the dairy products, dietary supplements or food additives, the active ingredient may include not only Lactobacillus brevis HD-01, but also commonly added ingredients in food preparation, for example, proteins, carbohydrates, fats, Nutrients, seasonings and flavoring agents.

Examples of carbohydrates used in food production include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And polysaccharides such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. As flavoring agents, natural flavoring agents [tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used.

Health functional food means that when ingested to bring a particular effect on health, unlike general medicine has the advantage that there is no side effect that can occur when taking a long-term use of the drug as a food raw material. The health functional food of the present invention thus obtained is very useful because it can be consumed on a daily basis.

When formulating the health functional food in the form of powder, granules, tablets or capsules, it may further comprise suitable carriers, excipients and diluents commonly used in food preparation. The carriers, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline Cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. In addition to the above excipients, lubricants such as magnesium styrate and talc are also used.

The amount of Lactobacillus brevis HD-01 added in the dairy products, health functional foods and food additives is usually in the range of 0.01 to 50% by weight, preferably 0.1 to 20% by weight relative to dairy products, health functional foods and food additives. 1 × 10 ⁵ to 1 × 10 ⁹ CFU / g, preferably 1 × 10 ⁵ to 1 × 10 ⁸ CFU / g, based on the final consumed formulation.

The daily intake of Lactobacillus brevis HD-01 of the present invention through the dairy products, health functional foods, food additives is 1 × 10 ⁵ to 1 × 10 ⁸ CFU / kg, preferably 1 × 10 ⁵ to 1 × 10 ⁷ CFU / kg, intake can be taken once a day or divided several times.

Hereinafter, the present invention will be described in more detail with reference to preferred embodiments. It will be apparent, however, to those skilled in the art that these embodiments are for further explanation of the present invention and that the scope of the present invention is not limited thereby.

Experimental Example  1: Isolation of Lactic Acid Bacteria from Kimchi

Lactic acid bacteria were isolated through 13 kinds of kimchi obtained at home. To separate the lactic acid bacteria, each kimchi was suspended in saline, and the suspension was plated in MRS agar medium and incubated at 37 ° C. for 48 hours. The lactic acid bacteria formed on the MRS agar were selected based on the different morphology and passaged at 37 ° C. for 24 hours at MRS broth. Among the isolated strains, 49 species were selected based on the formation of gas during cultivation, and the selected strains were confirmed to be all strains that generate lactic acid through BCP agar.

Experimental Example  2: Determination of Alcohol Resolution of Lactic Acid Bacteria

For 49 kinds of lactic acid bacteria selected in Experimental Example 1, 65 kinds of lactic acid bacteria possessed by Maeil Dairy, and a total of 114 kinds of lactic acid bacteria, alcohol resolution was measured by the following method.

1. Preparation of coenzyme solution of lactic acid bacteria

After culturing the strain in MRS broth medium at 37 ℃ for 24 hours, the cells were recovered and washed with sterile 100mM potassium phosphate buffer (PBS, pH 7.4). The recovered cells were suspended in 100 mM potassium phosphate buffer and McFarland No. 3 levels were set. The solubilization of the prepared bacteria suspension was performed 5 times for 30 seconds while slowly cooling on ice to prevent overheating. The cell lysate obtained at this time is centrifuged at 13,000 rpm for 1 hour. The supernatant was taken and the crude enzyme solution was separated and used in a cryogenic freezer at -80 ° C.

2. Determination of Alcohol Dehydrogenase (ADH) Activity

In order to determine the alcohol degradability of each lactic acid bacteria, the ADH activity was measured using the coenzyme solution. To 1 ml of a solution in which ethanol was added to 0.1 M Glycine buffer (pH 9.6) containing 2.5 mM NAD ⁺ , 100 μl of the coenzyme solution of each lactic acid bacterium was reacted for 5 minutes at 25 ° C. in a water bath. After the reaction was completed, the absorbance was measured at 340 nm, 25 ° C. in the resulting NADH content in the reacted solution. Protein content of the coenzyme solution was measured through a protein quantification kit (Bio-Rad). The measured results were used to compare the activity of ADH through the amount of NADH produced during the reaction time per mg of protein in the crude enzyme solution.

Among the results of measuring the ADH activity of the 114 strains, the isolates and scientific names of the 8 lactic acid bacteria having excellent ADH activity are shown in Table 1 in the order of high ADH activity, and the order of high ADH activity values of each lactic acid bacterium is shown in Table 2 Shown in

Strain number Strain name Scientific name Separation source One Kimchi isolated lactic acid bacteria 11 Lactobacillus brevis Kimchi 2 Kimchi Isolated Lactobacillus 24 Lactobacillus brevis Kimchi 3 Fermented milk isolated strain 1 Lactobacillus brevis Domestic fermented milk 4 Isolated strain 019 Lactobacillus fermentum Infant feces 5 Lactobacillus Percentum KCTC 10713BP Lactobacillus fermentum Privately Owned Strains 6 Lactobacillus ruteri LC40 Lactobacillus reuteri Commercial strain 7 Kimchi Isolated Lactobacillus 30 Lactobacillus collinoides Kimchi 8 Separated strain D4-1 Lactobacillus fermentum Owned strains

Strain number ADH activity (NADH μM / min / mg protein) One 297 2 282 3 209 4 159 5 132 6 86 7 85 8 58

In general, ADH activity values were higher in isolates than commercial strains, followed by kimchi lactic acid bacteria 11, kimchi lactic acid bacteria 24, fermented milk strain 1, and strain 019. Kimchi isolated lactic acid bacteria 11 (strain 1) and 24 (strain 2) showed 42% and 35% higher ADH activities than fermented milk isolated strain 1 (strain 3), which had the highest ADH activity among commercial strains.

Experimental Example  3: Determination of acetaldehyde resolution

Even if the alcohol is decomposed into acetaldehyde, acetaldehyde cannot be decomposed and accumulated, which may cause acute hepatotoxicity, which may cause a hangover. Therefore, it is important to rapidly decompose the acetaldehyde simultaneously with the decomposition of the alcohol, Experimental Example 2 ALDH activity was measured using the lactic acid bacteria coenzyme solution of Experimental Example 2 in order to confirm the acetaldehyde degradability of the eight strains with high alcohol degradability.

Acetaldehyde and 4-methylpyrazole were added to 60 mM Sodium pyrophosphate buffer (pH 8.8) containing 1 mM NADP ⁺ to a final concentration of 100 μM (4-methylpyrazole is an inhibitor of ADH). 1 ml of the mixed solution and 100 μl of each lactic acid coenzyme solution were mixed and reacted at 25 ° C. for 5 minutes in a water bath. After the reaction was completed, the resulting NADPH content in the reacted solution was measured for absorbance at 340nm, 25 ℃. Protein content of the coenzyme solution was measured through a protein quantification kit (Bio-Rad). The measured results are shown in Table 3 comparing the activity of ALDH through the amount of NADPH produced during the reaction time per mg of protein of the crude enzyme solution.

Strain number ALDH activity (NADPH μM / min / mg protein) One 60 2 27 3 17 4 15 5 26 6 15 7 6 8 34

Experimental Example  4: Acid resistance  And My bile  Measure

In Experimental Examples 2 and 3, the kimchi isolated lactic acid bacteria 11 (strain No. 1), which had the best ADH activity and ALDH activity at the same time, were compared with the commercial strain to confirm whether the acid resistance and the bile resistance were secured.

For acid resistance experiment, 50mM, a phosphate buffer was prepared, and the sterilized solution was adjusted to pH 3.0 with 1N HCL solution. Inoculated with 1% of the bacteria incubated in MRS broth medium for 24 hours in the acid solution was measured in the initial inoculation and the number of viable cells incubated at 37 ℃ for 2 hours is shown in Table 4. The measured number of live bacteria was expressed as LogCFU / ml, and it was judged that the difference was less acid resistant.

Strain number
Log (CFU / ml) 0 hours 2 hours Difference One 8.16 7.08 1.08 3 8.10 6.96 1.14 5 8.24 7.13 1.11 6 8.45 7.23 1.21

Kimchi isolated lactic acid bacteria 11 (strain No. 1) was confirmed to have better acid resistance than those compared to the commercial strain.

In addition, in the test of the bile resistance, a solution in which 1% of Oxgall (Difco) was added to 50mM phosphate buffer (pH 7) was used as a medium for the test for bile resistance. Inoculated by inoculating 1% in the biliary medium cultured bacteria in MRS broth medium for 24 hours to measure the initial number of inoculation and the number of viable cells incubated at 37 ℃ for 6 hours is shown in Table 5. The measured viable cell number was expressed as LogCFU / ml, and it was judged that the difference was small and the bile-resistant strain.

Strain number
Log (CFU / ml) 0 hours 6 hours Difference One 8.19 7.15 1.04 3 8.19 6.93 1.26 5 8.31 7.22 1.09 6 8.42 6.97 1.45

Kimchi isolated lactic acid bacteria 11 (strain No. 1) was found to have superior bile resistance than those compared to commercial strains.

Experimental Example  5: Identification of Selected Lactic Acid Bacteria

After separating the kimchi lactic acid bacteria 11 (strain No. 1) selected through Experimental Examples 1 to 4 into a single colony, the morphological and biochemical characteristics were investigated in accordance with Burgess's manual of systematic bacteriology. Gram staining was performed. As a result, it was confirmed that the isolated strain is a Gram-positive strain and has a form of bacilli.

In addition, the strain was identified by the API system (API) (La Balme-les-Grottes, France). First, bacteria colonies were taken with sterile platinum and then suspended in 2 ml of sterile distilled water. Again, the bacteria solution was suspended in 5 ml of sterile distilled water at a concentration of McFalland Standard Solution No. 2 (MccFaland Standard Solution No. 2) provided by the API 50CH kit (BioMerieux, France). The supernatant was added to the liquid medium of the API 50CH kit and homogenized, and 200 μl of each of the 50 tubes of the API 50CH kit was inoculated. After spilling on the tube with mineral oil, it was incubated for 48 hours at 30 ℃. The culture was analyzed by the API system to confirm 49 carbohydrate fermentation patterns, and the results were identified by inputting the ATB identification computer system.

As a result, the kimchi isolated lactic acid bacteria 11 was found to be a strain belonging to the genus Lactobacillus having a carbohydrate fermentation pattern of Table 6.

Control group - D-Manno - Salincin - Gentio Vioz - Glycerol - L-Solbos - Cellobiz - D-Turanz _ Erythritol - Rannoz - Malts + D-rings - D-arabinose - Dissytol - Laktos - D-Tagatoz - L-arabinose + Inositol - Melibiose + D-fucose - Riboze + Mannitol - Saka Rose - L-fucose - D-xylose + Sorbitol - Trehalose - D-arabitol - L-xylose - D-mannoside - Inulin - L-arabitol - Adonitol - D-glucoside + Melrietto - Gluconate + Xyloside - Glucosamine - D-Rapinoz - 2-Setto-gluconate - Galactos + Amigalline - Amidon - 5-Seto-gluconate + D-glucose + Arbutin - Glycogen - D-Fructose + Esculin + Xylitol -

In addition, the nucleotide sequence of 16S rDNA of the kimchi isolated lactic acid bacteria 11 was analyzed according to a conventional method known in the art. As a result, the 16S rDNA nucleotide sequence of the kimchi isolated lactic acid bacteria 11 (SEQ ID NO: 1) was found to be 99% identical to the 16S rDNA nucleotide sequence of Lactobacillus brevis.

Accordingly, the present inventors named Kimchi Lactobacillus 11 as "Lactobacillus brevis HD-01" and deposited it on January 4, 2012 at the National Institute of Agricultural Science, Agricultural Genetic Resources Center [Accession Number: KACC91701P].

Experimental Example  6: Animal testing for alcohol resolution

Animal tests were performed to determine whether the Lactobacillus brevis HD-01 exhibited alcohol degradability in vivo.

Lactobacillus brevis HD-01 from rat of 5 weeks old It was determined whether ingestion affects the rate of alcohol degradation. Example Lactobacillus brevis HD-01 Lactobacillus concentrate (1 × ^{10 9} CFU / ml) was prepared and provided in 1 ml each time. As a control, the comparative example provided water instead of the lactic acid bacteria suspension. The experiment proceeded with forced oral administration of lactobacillus suspension or water twice daily for two weeks to the rats. Feed and water were free-fed but water-saved 1-2 hours prior to administration of the lactobacillus suspension. After 14 days of administration, 1.5 ml of 40% ethanol solution was forcibly administered orally, and blood was collected at 1 hour intervals for 1-6 hours after administration, and then ethanol analysis was performed using GC (Gas Chromatography). . Results for blood alcohol concentration are shown in FIG. 1.

Example in which Lactobacillus brevis HD-01 of the present invention was ingested for two weeks was confirmed that alcohol was rapidly decomposed after ingesting alcohol. In particular, in the comparative example, the blood alcohol concentration was increased to 232 mg / dl at 1 hour, but in the example, it was reduced to 102 mg / dl, which is 56% lower than that in the case, and ingestion of Lactobacillus brevis HD-01 markedly improved alcohol resolution. I could confirm that.

Manufacturing example  1: Preparation of Fermented Milk

Fermented milk was prepared using Lactobacillus brevis HD-01 of the present invention.

Crude milk (74.41%) and skim milk powder (6.45%) are mixed and cultured by adding culture seed and Lactobacillus brevis to culture medium ( Lactobacillus brevis : 1 × 10 ⁷ CFU / ml) was prepared. As a comparison target, a culture medium in which only the culture seed was added and cultured in the same mixed solution composition was prepared. The sugar solution was prepared, and the fermented milk product was prepared by mixing the respective culture solutions prepared above.

Experimental Example  7: Preservation test of fermented milk

The fermented milk of Preparation Example 1 was placed in a 130ml empty bottle to block the inlet through air, and then the storage test was performed at 5 ° C. and 15 ° C., respectively. The sensory properties of the two products were compared by storing them at 4 ℃ and 15 ℃ for 2 weeks, and Lactobacillus during the storage period for products containing alcohol-degrading strains. The viable counts of brevis , TA, pH and gas production were measured.

In the case of sensory function, there was no significant difference from the comparison product during the storage period, and the result of preservation test of the product containing the Lactobacillus brevis HD-01 of the present invention is shown in Table 7 below.

Days saved Acidity pH Gas generation L. brevis (CFU / ml) 0 day 5 ℃ 0.54 4.35 (25.0 ℃) ㅡ 1.30E + 07 Day 3 5 ℃ 0.56 4.32 (26.7 ℃) X 5.40E + 07 15 ℃ 0.625 4.16 (28.1 ℃) X 6.12E + 07 Day 6 5 ℃ 0.575 4.30 (15.3 ℃) X 2.95E + 07 15 ℃ 0.67 4.14 (18.7 ℃) X 7.80E + 07 10th day 5 ℃ 0.72 4.04 (26.1 ℃) X 3.20E + 07 15 ℃ 0.78 3.95 (25.1 ℃) X 8.20E + 07 Day 14 5 ℃ 0.75 3.92 (24.5 ℃) X 5.80E + 07 15 ℃ 0.86 3.84 (24.4 ℃) X 8.95E + 07

As a result of preservation test, acidity progressed to a little faster when stored at 15 ° C, but acid progressed slowly when stored at 5 ° C. Conventional Lactobacillus brevis has a strong gas-forming strain during cultivation, but the culture of the Lactobacillus brevis HD-01 of the present invention did not progress rapidly in the product, the production of gas in a sealed product It was thought that this strain could be used in dairy products because there was no problem.

Manufacturing example  2: health functional food ( Powder Manufacturing

Lactobacillus brevis HD-01 lyophilized powder (1 × 10 ⁹ CFU / g) 20 mg

Lactose 100 mg

Talc 10 mg

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

Manufacturing example  3: Preparation of health functional food (tablet)

Lactobacillus brevis HD-01 lyophilized powder (1 × 10 ⁹ CFU / g) 10 mg

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above components, tablets are prepared by tableting according to the usual preparation method of tablets.

Manufacturing example  4: Health functional food ( Capsule Manufacturing

Lactobacillus brevis HD-01 lyophilized powder (1 × 10 ⁹ CFU / g) 10 mg

3 mg of crystalline cellulose

Lactose 14.8 mg

Magnesium Stearate 0.2 mg

The above components are mixed according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.

Manufacturing example  6: Health functional food Mixed powder)  Produce

Lactobacillus brevis HD-01 lyophilized powder (1 × 10 ⁹ CFU / g) 1 g

Vitamin mixture quantity

70 [mu] g of vitamin A acetate

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

0.15 mg of vitamin B2

Vitamin B6 0.5 mg

0.2 [mu] g vitamin B12

Vitamin C 10 mg

Biotin 10 μg

Nicotinic acid amide 1.7 mg

50 ㎍ of folic acid

Calcium pantothenate 0.5 mg

Mineral mixture quantity

1.75 mg of ferrous sulfate

0.82 mg of zinc oxide

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Secondary calcium phosphate 55 mg

Potassium citrate 90 mg

Calcium carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixture is comparatively mixed with a component suitable for a health functional food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above components may be mixed , Granules may be prepared and used in the manufacture of a health functional food composition according to a conventional method.

National Institute of Agricultural Science KACC91701P 20120104

&Lt; 110 > Maeil Dairies Co., Ltd. <120> Lactobacillus brevis with high alcohol dehydrogenase activity and          dairy products, health functional food and food additives          comprising the same <130> HPC-3048 <160> 1 <170> Kopatentin 2.0 <210> 1 <211> 1466 <212> DNA <213> Lactobacillus brevis <400> 1 cggggcggct gctatacatg cagtcgaacg agcttccgtt gaatgacgtg cttgcactga 60 tttcaacaat gaagcgagtg gcgaactggt gagtaacacg tgggaaatct gcccagaagc 120 aggggataac acttggaaac aggtgctaat accgtataac aacaaaatcc gcatggattt 180 tgtttgaaag gtggcttcgg ctatcacttc tggatgatcc cgcggcgtat tagttagttg 240 gtgaggtaaa ggcccaccaa gacgatgata cgtagccgac ctgagagggt aatcggccac 300 attgggactg agacacggcc caaactccta cgggaggcag cagtagggaa tcttccacaa 360 tggacgaaag tctgatggag caatgccgcg tgagtgaaga agggtttcgg ctcgtaaaac 420 tctgttgtta aagaagaaca cctttgagag taactgttca agggttgacg gtatttaacc 480 agaaagccac ggctaactac gtgccagcag ccgcggtaat acgtaggtgg caagcgttgt 540 ccggatttat tgggcgtaaa gcgagcgcag gcggtttttt aagtctgatg tgaaagcctt 600 cggcttaacc ggagaagtgc atcggaaact gggagacttg agtgcagaag aggacagtgg 660 aactccatgt gtagcggtgg aatgcgtaga tatatggaag aacaccagtg gcgaaggcgg 720 ctgtctagtc tgtaactgac gctgaggctc gaaagcatgg gtagcgaaca ggattagata 780 ccctggtagt ccatgccgta aacgatgagt gctaagtgtt ggagggtttc cgcccttcag 840 tgctgcagct aacgcattaa gcactccgcc tggggagtac gaccgcaagg ttgaaactca 900 aaggaattga cgggggcccg cacaagcggt ggagcatgtg gtttaattcg aagctacgcg 960 aagaacctta ccaggtcttg acatcttctg ccaatcttag agataagacg ttcccttcgg 1020 ggacagaatg acaggtggtg catggttgtc gtcagctcgt gtcgtgagat gttgggttaa 1080 gtcccgcaac gagcgcaacc cttattatca gttgccagca ttcagttggg cactctggtg 1140 agactgccgg tgacaaaccg gaggaaggtg gggatgacgt caaatcatca tgccccttat 1200 gacctgggct acacacgtgc tacaatggac ggtacaacga gtcgcgaagt cgtgaggcta 1260 agctaatctc ttaaagccgt tctcagttcg gattgtaggc tgcaactcgc ctacatgaag 1320 ttggaatcgc tagtaatcgc ggatcagcat gccgcggtga atacgttccc gggccttgta 1380 cacaccgccc gtcacaccat gagagtttgt aacacccaaa gccggtgaga taaccttcgg 1440 gagtcagccg tctaaggtga accagt 1466

Claims (7)

Lactobacillus brevis (Lactobacillus high resolution alcohol brevis ) HD-01 [Accession No. KACC91701P].
According to claim 1, Lactobacillus brevis ( Lactobacillus) characterized in that the activity of alcohol dehydrogenase and acetaldehyde dehydrogenase is excellent at the same time brevis ) HD-01 [Accession No. KACC91701P].
A dairy product containing Lactobacillus brevis HD-01 [Accession No .: KACC91701P] excellent in alcohol resolution of claim 1.
4. The dairy product of claim 3 wherein the dairy product is fermented milk, butter, cheese or milk powder.
Claim 1, Lactobacillus brevis ( Lactobacillus brevis ) HD-01 [Accession No .: KACC91701P] excellent alcohol decomposing health functional food for preventing or hangover.
According to claim 5, wherein the health functional food is an alcoholic liver disease prevention or hangover health functional food, characterized in that in the form of powder, granules, tablets or capsules.
A food additive containing Lactobacillus brevis HD-01 [Accession No .: KACC91701P] excellent in alcohol resolution of claim 1.

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