TWI405849B - Lactic acid bacteria having a function of lowering uric acid in blood - Google Patents

Abstract

Various lactic acid bacteria were cultured in the presence of a purine body and the consumed amount of the purine body, the produced amount of degradation products of the purine body were measured, and a plurality of lactic acid bacteria which has a remarkable degradation ability of purine body were selected. Any of the lactic acid bacteria which were judged to have a high degradation ability of purine body by the selection was orally administered to rats raised with a purine body-containing feed, the general status and the blood uric acid level were measured, and the effect of administration of lactic acid bacteria on the serum uric acid level was studied. As a result, lactic acid bacteria: Lactobacillus gasseri OLL2959 and Lactobacillus oris OLL2779 were found having a significant effect in reducing the serum uric acid level.

Description

Lactic acid bacteria having the effect of lowering uric acid value in blood

The present invention relates to a lactic acid bacterium having an effect of lowering uric acid value in blood and an application thereof, and a food or a pharmaceutical product containing the lactic acid bacteria for preventing and/or treating hyperuricemia.

Hyperuricemia is caused by environmental factors (living habits) and genetic factors, which leads to a decrease in uric acid excretion or an excess of uric acid, and a state in which blood uric acid is too high. Hyperuricemia may also have no symptoms, but is caused by severe complications such as gout, renal insufficiency, urinary calculi, and arteriosclerosis. A representative complication of hyperuricemia is gout, and the main symptom appears as acute arthritis accompanied by severe pain. In the past, gout was called "imperial disease", and it was called "spreading disease" because of frequent and large intake of meat, fish, or alcohol. In recent years, there has been a tendency to increase cases every year due to changes in dietary life. The current number of gout patients in Japan is 300,000 to 400,000, and the number of patients with hyperuricemia is estimated to be 6 million. Therefore, the public is also highly concerned about the prevention or treatment of hyperuricemia.

The prevention or treatment of hyperuricemia is the control of blood uric acid value by a combination of food therapy, exercise therapy, pharmaceuticals, and combinations thereof. In particular, it is one of the most frequently selected methods for preventing or treating hyperuricemia by limiting caloric intake. However, it is quite difficult to strictly limit calorie intake for a long time. It has been published that the method of improving this condition can be decomposed by oral intake. (or commonly known as purin), lactic acid bacteria, yeast, and other microbial medicines or drinks In the food, the sputum ingested by the diet is decomposed in the intestine, and the absorption in the body is reduced, and the serum uric acid value is lowered (Patent Document 1 and Non-Patent Document 1). Since ancient times, lactic acid bacteria have been frequently used in foods or pharmaceuticals, and their safety to humans is high. Lactic acid bacteria intake has few side effects, which is an effective method for treating or preventing hyperuricemia. Further, as described above, the first choice for the prevention or treatment of hyperuricemia is food therapy, and lactic acid bacteria which can control uric acid value through food intake are highly practical and useful novel hyperuricemia prevention and/or treatment methods. However, the lactic acid bacteria which can decompose sputum disclosed in the above documents include Lactobacillus fermentum and Lactobacillus pentosus. These lactic acid bacteria produce gas, and it is not an appropriate bacteria from the viewpoint of application of foods and drinks or pharmaceuticals. Kind.

Patent Document 1: WO2004/112809

Non-Patent Document 1: Japanese Agrochemical Society Homepage Agronomic Chemistry Annual Conference Speech Presentation Database ( http://isbba.bioweb.ne.jp/jsbba_db/index.html ) "National Agrochemical Society 2004.03.30 General Lecture, Chi Yongwu, Jiu Mi Cun Hui: The effect of lactic acid bacteria on blood uric acid value in rats with dietary hyperuricemia

In view of the above circumstances, the problem to be solved by the present invention is to provide a food or food product for use in prevention and/or treatment of high uric acid. The lactic acid bacteria of the bacterium also provides a composition for preventing and/or treating hyperuricemia using the above lactic acid bacteria.

In order to solve the above problems, the inventors of the present invention have made repeated efforts to study. First, various lactic acid bacteria were cultured in the presence of inosine and guanosine, and the above-mentioned nucleoside consumption and decomposition products (hypoxanthine, guanine) production amount were measured, and the selection was remarkable. A plurality of lactic acid bacteria that have the ability to decompose nucleosides. A lactic acid bacterium having a high nucleoside-decomposing ability based on the above selection is orally administered to a rat bred with a sputum feed, and the general condition and serum uric acid value of the rats are measured, and the lactic acid bacteria are administered for serum uric acid. The impact of the value. As a result, it was found that the lactic acid bacteria having a significant inhibitory effect on the increase in serum uric acid value were Lactobacillus oryzae OLL2779 strain and Lactobacillus gasseri OLL2959 strain. Further, the inventors of the present invention used the above-described lactic acid bacteria to prepare a yogurt to confirm whether or not the above lactic acid bacteria are suitable for food processing applications containing yoghurt. The lactic acid bacterium of the present invention has an inhibitory action against an increase in serum uric acid value, and can be effectively utilized for a pharmaceutical having a preventive and/or therapeutic use for high uric acidosis and gout. Further, the inhibitory effect of the increase in serum uric acid value of the lactic acid bacteria of the present invention is also confirmed by an experiment for oral administration, and it is confirmed that it is suitable for use in actual food processing, and it has a remarkable high use as a food application. Accordingly, the present invention relates to a lactic acid bacterium capable of preventing and/or treating hyperuricemia and an application thereof, and in particular, the present invention provides the following invention.

(1) A Lactobacillus lactic acid bacterium having a cockroach 呤 Decomposition ability, no gas production capacity, (2) Lactobacillus gasseri, as shown in (1) above, is Lactobacillus gasseri.

(3) A Lactobacillus oris having a decomposing ability, and (4) a Lactobacillus lactic acid bacterium as described in the above (2), which is a Lactobacillus gastrin OLL2959 strain (Lactobacillus gasseri OLL2959) : NITE BP224), (5) Lactobacillus lactic acid bacteria as described in (3) above, which is Lactobacillus kawaii OLL2779 (Lactobacillus oris OLL2779) (Accession No.: NITE BP-223), (6) one for blood A food or drink having an inhibitory effect on an increase in uric acid value, comprising the lactic acid bacterium according to any one of the above (1) to (5), the lactic acid bacteria-containing substance and/or a processed product thereof, and (7) a uric acid value in blood A medicinal product having an inhibitory effect, comprising the lactic acid bacterium according to any one of the above (1) to (5), the lactic acid bacteria-containing substance and/or a processed product thereof, and (8) one for prevention and/or treatment A uric acid bacterium according to any one of the above (1) to (5), the lactic acid bacteria-containing substance and/or a processed product thereof, and (9) a drug for prevention and/or treatment A food or drink of hyperuricemia, which contains as set forth in any one of the above (1) to (5) Lactic acid bacteria, lactic acid bacteria-containing composition described above and / or a processed material (10) an amount of one purine by the method of inhibition of food intake, wherein The lactic acid bacteria according to any one of the above (1) to (5), the lactic acid bacteria-containing substance and/or the processed product thereof, and (11) a method for suppressing an increase in uric acid value in blood, which is characterized in that The lactic acid bacteria according to any one of the above (1) to (5), the lactic acid bacteria-containing material and/or a processed product thereof, and (12) a method for producing a food or drink having a reduced amount of sputum, which comprises a raw material of a food or drink or The intermediate product is in contact with the lactic acid bacteria according to any one of the above (1) to (5), the lactic acid bacteria-containing substance and/or the processed product thereof, and (13) one for prevention and/or treatment of hyperuricemia And a method of any of the diseases or symptoms of a group consisting of gout, renal dysfunction, urinary calculi, and arteriosclerosis, characterized by administering as set forth in any one of the above (1) to (5) And a lactic acid bacterium according to any one of the above (1) to (5), wherein the lactic acid bacteria-containing substance and/or a processed product thereof is used. A food or drink and/or a pharmaceutical product which suppresses an increase in uric acid value in blood of a subject to be administered.

The present invention relates to a lactic acid bacterium of the genus Lactobacillus which has the ability to decompose hydrazine and has no gas generating ability. The present invention is a Lactobacillus lactic acid bacterium (hereinafter referred to as "the Lactobacillus lactic acid bacterium of the present invention" having the cockroach decomposition ability and having no gas generating ability, which was first discovered by the present inventors.

Lactobacillus is a genus of bacteria represented by lactic acid bacteria and contains more than 80 species of bacteria. Examples of strains contained in the genus Lactobacillus are Lactobacillus delbrueckii subsp. burgalicus, Lactobacillus delbrueckii subsp. lactis, Lactobacillus subfamily (Lactobacillus). Paracasei subsp.paracasei), Lactobacillus helveticus, Lactobacillus helveticus subsp. jugurti, Lactobacillus acidophilus, Lactobacillus acidophilus crispatus, Lactobacillus amylovorus, Lactobacillus gallinarum, Lactobacillus gasseri, Lactobacillus oris, Lactobacillus casei subsp. rhamnosus, about Lactobacillus johnsonii, Lactobacillus fermentum, Lactobacillus brevis. The lactic acid bacterium of the genus Lactobacillus of the present invention has a cockroach-decomposing ability, but has no gas-producing ability, and is preferably any one of the genus Lactobacillus, and is preferably Lactobacillus gasseri.

The quinone-based nucleic acid constituents are supplied to the living body via de novo synthesis, salvage pathways, nuclear proteins in the prey, and the like, and unnecessary sputum can be excreted by the liver. Uric acid is the ultimate metabolite of cockroaches in humans, higher primates, birds, reptiles and other animals.

By hydrazine in the present specification is meant a compound having a fluorene structure. Representative examples of sputum are, for example, purine nucleotides (adenine nucleotides, deoxyadenosine nucleotides, guanine nucleotides, deoxyguanine nucleotides), nucleosides (adenosine, deoxygenation) Adenosine, guanosine, deoxyguanosine), purine bases (adenine, guanine), purine-containing oligonucleotides and polynucleotides. In addition to the nucleic acid constituents, the purine base is also a component of various living organisms such as ATP, GTP, cAMP, cGMP, coenzyme A, FAD, and NAD. The reference in this specification is not limited to those having a skeleton, and all the components in the above-mentioned living body are also included.

The lanthanide in the body is metabolized in the form of uric acid. The metabolic pathway of the uric acid form of sputum is well known. The AMP is treated with 5'-nucleotidase to form adenosine, and the adenosine is inosine to become hypoxanthine. The GMP is treated with 5'-nucleotidase to form guanosine, and then guanine is formed. Hypoxanthine is treated with xanthine oxidase, and guanine is treated with guanine deaminase, which is metabolized to xanthine, which is then treated with xanthine oxidase to form uric acid.

The so-called cockroach decomposition ability of the present invention has at least one hydrazine decomposition ability regardless of whether the decomposition product has a sacral backbone. That is, the ability to decompose sputum to form a compound having no sacral stem, or to decompose sputum to form other sputum (a compound having a sacral backbone) is included in the so-called sputum decomposition ability of the present invention.

The so-called Lactobacillus lactic acid bacteria of the present invention can be isolated by a known method. For example, bacteria obtained by cultivating feces from mammals such as humans are separated from the shape and physiological characteristics of the cultured bacteria. A bacterium belonging to the genus Lactobacillus is tested for its ability to decompose and produce gas. It is selected from the genus Lactobacillus which has the ability to decompose sputum and has no gas producing ability and can be isolated. The detection of the decomposing ability and the detection of the gas generating ability can be carried out by a conventional method, and can be carried out, for example, by the method of the present embodiment.

The culture system of the Lactobacillus lactic acid bacteria of the present invention is preferably a medium suitable for general Lactobacillus culture, and a carbon source containing glucose, lactose, galactose, fructose, sea bath sugar, sucrose, mannose, cellobiose or the like can be used. A medium for extracting inorganic substances such as meat extract, peptone, yeast extract, casein, whey protein, or the like, and inorganic nutrients such as magnesium sulfate, iron sulfate, and manganese sulfate. A preferred example is Lactobacilli MRS broth (produced by Difco Co., Ltd.). The culture condition is a condition in which the lactic acid bacteria in the intestine can be propagated, and is not particularly limited. Preferred conditions are, for example, pH 5.0 to pH 8.0, temperature 20 ° C to 45 ° C, and more preferable conditions, an anaerobic environment, pH 5.0 - pH 7.0, temperature 30 ° C - 40 ° C.

As described below, the inventors of the present invention orally administered the Lactobacillus lactic acid bacteria of the present invention to a model animal, and confirmed the inhibitory effect of the lactic acid bacteria on the increase in uric acid value in the blood. Therefore, since the Lactobacillus lactic acid bacterium of the present invention can suppress an increase in uric acid value in blood, it can be applied to prevent and/or treat hyperuricemia. Further, by using the cockroach decomposition ability of the lactic acid bacterium of the lactic acid bacterium of the present invention, it is conceivable to manufacture a food having a reduced amount of mash.

A specific example of the "Lactobacillus lactic acid bacterium having a sputum-decomposing ability and a gas-producing ability" of the present invention is a specific L. lactis OLL2959 strain (Lactobacillus gasseri OLL2959) having the accession number: NITE BP-224. The present inventors have examined the decomposition ability of various lactic acid bacteria and The presence or absence of gas production ability revealed that Lactobacillus lactic acid bacteria named Lactobacillus gasseri OLL2959 had the ability to decompose and produce gas. And in vivo test, it was found that Lactobacillus kawaii OLL2959 strain (Lactobacillus gasseri OLL2959) obviously inhibited the increase of uric acid value in blood. The present inventors deposited the above-mentioned strains in the patent microbial deposit center of the independent administrative legal person product evaluation technology base institution. The following is the content of the specific deposit:

(1) Depository authority: Patent Microbiology Depository Center for Product Evaluation Technology Basement Organization of Independent Administrative Corporation (Address: 2-5-8, Post Code, 292-0818, Kishozu, Kisarazu, Chiba Prefecture, Japan)

(2) Original registration date: March 31, 2006 based on the Budapest Treaty, transferred by the domestic depository management day: November 21, 2007

(3) Deposit number: Lactobacillus gasseri OLL2959 strain (registration number NITE BP-224)

The above-mentioned strains are additionally deposited in the Food Industry Development Research Institute of the Republic of China. The deposit number is BCRC 910379, and the deposit date is December 24, 1996.

Further, the present invention relates to Lactobacillus oris having a decomposing ability. The present invention is based on the first discovery by the present inventors that Lactobacillus oris (hereinafter referred to as "Lactobacillus brevisii" of the present invention) having a decomposing ability.

The Lactobacillus brevis strain of the present invention is isolated by the same method as described above. ‧ The Lactobacillus lactic acid bacterium of the present invention is cultured. Further, the Lactobacillus brevis of the present invention is also useful for treating and/or preventing hyperuricemia because it has an inhibitory effect on the increase in uric acid value in blood.

A specific example of the "Lactobacillus oris having a decomposing ability" of the present invention is a specific Lactobacillus oris OLL2779 strain (Lactobacillus oris OLL2779) having the accession number: NITE BP-223. The inventors of the present invention have found a lactic acid bacterium having a sputum-decomposing ability among various lactic acid bacteria, Lactobacillus oryzae OLL2779 strain (Lactobacillus oris OLL2779), and confirmed that the Lactobacillus oryzae OLL2779 strain (Lactobacillus oris OLL2779) has apparently passed through an in vivo test. Inhibition of increased uric acid levels in the blood. The present inventors deposited the above-mentioned strains in the patent microbial deposit center of the independent administrative legal person product evaluation technology base institution. The following is the content of the specific deposit:

(1) Depository authority: Patent Microbiology Depository Center for Product Evaluation Technology Basement Organization of Independent Administrative Corporation (Address: 2-5-8, Post Code, 292-0818, Kishozu, Kisarazu, Chiba Prefecture, Japan)

(2) Original registration date: March 31, 2006 based on the Budapest Treaty, transferred by the domestic depository management day: November 21, 2007

(3) Deposit number: Lactobacillus oris OLL2779 strain (registration number: NITE BP-223)

The above-mentioned strain was deposited in the Food Industry Development Research Institute of the Republic of China, and the deposit number is BCRC 910378. The deposit date is December 24, 1996.

The Lactobacillus lactic acid bacterium of the present invention and the Lactobacillus oral bacterium of the present invention can be used for a food or drink or a pharmaceutical product which suppresses an increase in uric acid value in blood, or a pharmaceutical or food or drink which can prevent and/or treat hyperuricemia.

When the Lactobacillus lactic acid bacterium of the present invention and the Lactobacillus acidophilus of the present invention are used as foods and drinks, the properties and types thereof are not limited, and functional foods, specific health foods, health foods, and foods for care are preferable. Fruits, lactic acid bacteria beverages, cheese and yogurt, and other seasonings are also good. The shape of the food and drink is also unlimited, and it is solid, liquid, flowing, jelly, lozenge, granule, capsule, etc., and the shape of the food and beverage that is generally on the market is acceptable. The production of the above-mentioned foods and drinks is carried out by the usual methods of the practitioner. When manufacturing the above-mentioned food and beverage products, it is limited to those who do not affect the growth and reproduction of lactic acid bacteria, and may add sugars, proteins, lipids, dietary fibers, vitamins, and trace metals (manganese sulfate, zinc sulfate, magnesium chloride, potassium carbonate, etc.) and spices required for living organisms. And other matching additives.

The lactic acid bacterium of the genus Lactobacillus of the present invention, the lactic acid bacterium of the present invention, and the contents of the lactic acid bacteria and/or the processed material thereof (for example, culture, concentrate, paste, spray dried product, lyophilized product, vacuum) The dried product, the dried product of the drum, the liquid, the diluted product, and the crushed product are used in the processing of the general foods and drinks containing the above-mentioned dairy products and fermented milk, and the manufacture of dairy products and fermented milk such as Yogurt or cheese. It is manufactured by a starter or the like. The initial flora is limited to the Lactobacillus lactic acid bacteria and the present invention of the present invention. The growth and proliferation of Lactobacillus brevis is not inhibited, and it is also possible to mix with other microorganisms if it is not restricted to the production of dairy products. For example, it is also preferable to mix with Lactobacillus delbruekii subsp. bulgaricus, Streptococcus thermophilus, Lactobacillus acidophilus, and the like, which are the main species of lactic acid bacteria. In addition, it can also be mixed with a strain for general use or cheese as a starting flora. The production of the dairy product and the fermented milk by the above-mentioned starting flora can be carried out according to a usual method. For example, a dairy product or a dairy product which is heated, mixed, homogenized, and sterilized and cooled is mixed with the above-mentioned starting flora, fermented and cooled to produce an original flavor.

The lactic acid bacterium of the genus Lactobacillus of the present invention and the lactic acid bacterium of the present invention may be mixed with a physiologically acceptable carrier, excipient, or diluent, etc., and may be administered as a pharmaceutical composition by oral or parenteral administration, preferably. The method of investment is administered orally. The orally administered preparation may be in various known dosage forms, and may be, for example, a granule, a powder, a lozenge, a pill, a capsule, a liquid, a syrup, an emulsion, a suspension, a throat, or the like. Further, the enteric preparation prepared by a known method by a practitioner is not affected by gastric acid, and the Lactobacillus lactic acid bacteria of the present invention can be efficiently operated into the intestine.

By using the lactic acid bacteria of the lactic acid bacterium of the present invention and the medicinal products and foods and beverages produced by the lactic acid bacillus of the present invention, it is expected that the effect of the same bacteria in the food and beverage can be exerted to suppress the increase in uric acid value in the blood and the high uric acid. Prevention and/or treatment of blood.

Further, the Lactobacillus lactic acid bacterium of the present invention and the present invention can be used. The decomposition ability of Lactobacillus brevis is used to produce foods and drinks with reduced sputum. The method for producing a food or drink having a reduced amount of the present invention includes the step of bringing the Lactobacillus lactic acid bacteria of the present invention or the Lactobacillus oral bacterium of the present invention into contact with a raw material or an intermediate product of a food or beverage. Through this step, the amount of rhenium contained in the raw material or intermediate product can be effectively reduced. The above steps are preferably carried out under the conditions in which the Lactobacillus lactic acid bacteria of the present invention and the Lactobacillus oral bacterium of the present invention survive. In the manufacturing method of the present invention, the above steps are included in the usual steps of the pulverizing step, the mixing step, the drying step, the sterilizing and filling steps, and the like. The foods and drinks produced by the method of the present invention are not limited in terms of their properties and types, such as functional foods, specific health foods, health foods, and foods for care, and general foods such as foods classified as hobby goods are also preferred. A patient who needs to limit the disease or symptoms of sputum intake is particularly useful as a daily or supplementary food or drink for preventing the above diseases or symptoms. In the method of the present invention, it is necessary to limit the amount of sputum intake, or it is desirable to limit the amount of sputum intake, and it is possible to ingest a food having a large amount of food and enjoy the nutrients and flavor of the food.

Moreover, all of the prior art documents cited in the specification can be included in the present specification as a reference to the present specification.

Example

Hereinafter, the present invention will be more specifically described based on examples of the invention. However, the invention is not limited to the embodiments described below. Further, in the examples, the strain whose strain name is JCM is a standard strain obtained by the Microbial Materials Development Laboratory of the Biological Resource Center of the Institute of Physical and Chemical Research, Institute of Physical and Chemical Research, and the strain name is recorded by ATCC. The strain contained is a standard strain obtained by the American Type Culture Collection, and the strain whose strain name is MEP is a strain possessed by Meiji Dairy Co., Ltd.

[Example 1] In vitro experimental method relating to the action of lactic acid bacteria to reduce uric acid

Various lactic acid bacteria were studied for the presence or absence of their decomposition ability by the following method. Various lactic acid bacteria (bacteria) are made of Difco Lactobacilli MRS Broth (made by BD) as a medium, and an oxygen sorbent "anaerobic culture package" (manufactured by Mitsubishi Gas Co., Ltd.). It was placed in a closed container and anaerobic cultured overnight at a temperature of 37 °C. The cultured suspension of the cells was centrifuged at 3000 rpm for 10 minutes at a temperature of 4 ° C, and the isolated cells were recovered by precipitation (serum collection).

This cell was prepared with a suspension of 1×10 ⁹ CFU/mL of the cell suspension in 0.1 M sodium citrate buffer.

After preparing various cell suspensions, inosine and guanosine were separately added to various cell suspensions to a concentration of 1.25 mM. These cell suspensions were placed in a 37 ° C thermostat and shake cultured at a horizontal speed of 140 rpm for 30 minutes or 2 hours.

For the cell suspension (reaction solution) after vibration culture, 5-bromouracil was used as an internal standard, and the amount of nucleoside consumption and the amount of nucleoside-degrading base (hypoxanthine and guanine) were determined by HPLC. . In 780 μL of the mobile phase A, 200 μL of the reaction solution and 20 μL of 5-bromouracil (1.6 mg/mL) as an internal standard were added and mixed. After the mixture was filtered through a filter (pore size: 0.45 μm), 50 μL of the filtrate was poured into HPLC. HPLC The body operating conditions are as follows.

HPLC: Waters alliance 2690

Column: CAPCELL PAK C ₁₈ SG120, particle size 5μm, column size 4.6×250mm (Shiseido)

Mobile phase: A: 25 mM KH ₂ PO ₄ (0.1% methanol) B: 25 mM KH ₂ PO ₄ (0.1% methanol) / methanol (75: 25)

Gradient A/B(min): 100/0(0)-100/0(10)-20/80(20)-20/80(25)-100/0(26)-100/0(40)

Verifier: Photodiode array (Waters 996) detected wavelength 254nm

Flow rate: 1mL/min

Column temperature: normal temperature

The results are shown in Figures 1 to 3. The quantification of each compound was based on the peak area values in the HPLC plot. Further, the decomposition rates of the first and second figures are calculated according to the following equation.

Decomposition rate = 100 - (inosine or guanosine amount / blank control group inosine or guanosine amount) × 100

In addition, the calculation method of the result of Fig. 3 is in accordance with the following formula.

(second dose of jaundice + amount of guanine) / 5 - bromo uracil amount

From the results of Figs. 1 to 3, lactic acid bacteria which were judged to have significant nucleoside decomposition ability were selected.

[Example 2] In vivo test method relating to uric acid lowering action of lactic acid bacteria

According to the method described in the prior literature (Non-Patent Document 1), a model animal of causative hyperuricemia is produced, and the effect of the microorganism (lactic acid bacteria) on the serum uric acid value of the animal is examined. In the above method, specifically, a mixed bait containing 2.5% by weight of potassium phenylaminocarbonate and 1.0% by weight of RNA was prepared, and the rats were ingested with the mixed baits, and the uric acid value in the blood of the negative group and the control group after ingestion was compared. It is known that the oral administration of the uric acid-producing blocker allopurinol to the model animal of the present method significantly inhibits the uric acid value in the blood of the model animal (Food Function Research News (No. 14), 2005 3 Issued on the 9th of June, (Mechanian Cleantec Environmental Inspection Center, http://www.m-cleantec.com/gizyutu/news_0503.htm ). The above method showing the effectiveness evaluation system for hyperuricemia foods is effective.

[2-1 Materials and Experimental Procedures]

[microorganism]

Those who have high nucleoside-dissolving ability in the above-mentioned in vitro test are Lactobacillus fermentum MEP181504 (hereinafter referred to as "Lactobacillus" or "L.") and Lactobacillus sp. MEP181507 (L. brevis MEP181507). There were 5 strains of Lactobacillus sp. JCM8787 strain (L. gasseri JCM8787), Lactobacillus delbrueckol OLL2959 strain (L. gasseri OLL2959), and Lactobacillus acidophilus OLL2779 strain (L. oris OLL2779). Various lactic acid bacteria were prepared into the same cell suspension as in the in vitro test. The bacterial suspension was orally administered to the rats in an amount of 1 × 10 ⁹ CFU/10 mL / kg.

[experimental animal]

Rats (Wister SPF, male, 7 weeks old) were used. The breeding (domestication and test) process uses a plastic cage for rats, and each cage contains one rat. The light and dark cycle is from 7 am to 7 pm (12 hours) in the Ming Dynasty.

[prepared breeding (domestication) and grouping]

After the experimental animals were moved in, they were pre-incubated for 1 week (domestication). The domestication process can be freely fed, with AIN-93G (Oriental Yeast Industry Co., Ltd.) as the feeding ground (feed) and tap water as the drinking water. The rats that were prepared for breeding (after 7 days, 8 weeks, and 0 days) were collected from the tail vein in the morning, not in the case of non-fasting. The blood was allowed to stand at room temperature for 30 minutes or more, and the serum was separated by centrifugation at 10,000 rpm for 10 minutes, and the uric acid value in the serum was measured by the phosphotungstic acid method.

The grouping was performed in such a manner that each group had the same serum uric acid value. In the experiment, 5 rats in group 1 were set as the negative group (groups 1 and 5), the control group (groups 2 and 6), and the bacterial administration group (groups 3, 4, 7-9). A total of 9 groups. The group name, the prey, the dosing (dosing amount), and the number are as follows.

‧ Negative group (Groups 1 and 5): Give "AIN-93G" prey, cast "physiological saline" (10mL/Kg), 5

‧ Control group (Groups 2 and 6): The "AIN-93G mixed with 2.5% by weight of phenylaminocarbonate and 1.0% by weight of RNA" was administered, and "physiological saline" (10 mL/kg) was administered. only

‧ Bacterial administration group (Groups 3, 4, 7 to 9): Each group was given a "bait of AIN-93G mixed with 2.5% by weight of potassium phenylaminocarbonate and 1.0% by weight of RNA", and a group of five. The amount of cells administered in each group and the amount administered were as follows: Group 3 was administered "Lactobacillus fermentum MEP181504 strain (L. fermentum MEP181504) suspension (1 × 10 ⁸ CFU/mL)" (10 mL/kg), 4th The group was administered with "Lacillus brevis MEP181507 strain (L. brevis MEP181507) suspension (1 × 10 ⁸ CFU/mL)" (10 mL / kg), and the seventh group was administered with "L. oris OLL2779 strain (L. oris OLL2779). Suspension (1×10 ⁸ CFU/mL)” (10 mL/kg), Group 8 was administered with “L. gasseri JCM8787 suspension (1×10 ⁸ CFU/mL) (10 mL/kg) and the ninth group were administered "L. gasseri OLL2959 strain (L.gasseri OLL2959) suspension (1 × 10 ⁸ CFU/mL)" (10 mL/kg).

[Formal breeding (test)]

From the next day after grouping, the "AIN-93G" feed (negative group) and the "AIN-93G + aniline potassium carbonate + RNA" feed (control group, bacterial administration group) were given to the bait feeder. Free to eat on the 8th. The start date of the feed of the above feed is taken as the first day, and the number of days is counted in the order of the day. The "AIN-93G + potassium aniline potassium carbonate + RNA" feed contained 2.5% by weight of potassium phenylaminocarbonate (100 g, ALDRICH product) and 1.0% by weight of RNA (500 g, product of MP Biomedicals. Inc.). The experimental animals of the bacterial administration group were orally administered with 1 x 10 ⁹ CFU/10 mL/kg by the above-mentioned bacterial suspension. The negative suspension group and the control group were not administered with the suspension of the cells, but the physiological saline was forcibly administered orally with 10 mL/kg.

[Measurement and inspection, etc.]

‧General status observation and weight measurement

All (all groups) observe the general state from the 1st to the 8th consecutive day, and the 0th, 1st, 5th, and 8th are in the afternoon. The weight is measured regularly from the first 9 to 10 o'clock.

‧ Determination of food intake and water intake

The whole (all groups) measured the food intake and the amount of water intake at the first day (set value), the fifth day (residual value, the set value), and the eighth day (residual value) at 9:10 pm.

‧ Blood collection and biochemical examination

All (all groups) were collected from the tail vein on day 0 (noon), day 2 (after 1 hour of administration), day 5 (after 1 hour of administration), and day 8 (before administration). The blood taken was centrifuged at 10,000 rpm for 10 minutes to separate the serum, and the uric acid value in the serum was measured by the phosphotungstic acid method. As described above, the uric acid value in the serum on the day of the measurement was measured on the 0th day, and then separately measured.

‧ necropsy and biochemical examination

All (all groups) were subjected to blood collection from the tail vein on the 8th day, and the suspension of the cells was orally administered. One hour later, after anesthesia with pentobarbital (trade name: Nembutal) (pentobarbital 40 mg/kg), blood was collected from the abdominal aorta. The blood taken was centrifuged at 3000 rpm for 15 minutes to remove the serum, and creatinine, uric acid, and urea nitrogen in the serum were measured.

‧ Determination of organ weight

The kidneys of the rats were removed and the wet weight was measured.

[Statistical processing]

The results were shown as mean ± standard deviation, and the control group and each bacterial cell administration group were compared. The numerically-defined test value dispersion ratio is performed by F-test, and the dispersion is measured by Student's T test, and the unequal dispersion is analyzed by Aspin-Welch T test. . Statistical processing was performed using Excel 2004 for statistical analysis with a minimum significant level of 5% on both sides.

[2-2 results]

The results of the general state are shown in Table 1 (L. fermentum and L. brevis) and Table 2 (L. oris and L. gasseri), and the changes in serum uric acid values are shown in Figs. 4 and 5.

The administration of various lactic acid bacteria can cause a decrease in serum uric acid value in Fig. 5, and the L. oris OLL 2779 strain administration group and the L. gasseri OLL 2959 strain administration group are considered to have statistically significant differences. The general status shows that no matter what group of kidney function (creatinine value, serum urea nitrogen, kidney weight), there is no problem, and there is no problem in body weight, food intake, and water intake. In addition, as seen from the general state, no significant difference was observed between various lactic acid bacteria. The scientific characteristics of L. oris OLL 2779 strain and L. gasseri OLL 2959 strain are shown in Table 3.

[Example 3] Production of fermented milk

(Production Example 1 of fermented milk)

The original yoghurt was prepared by Lactobacillus kawaii OLL2959 strain (L. gasseri OLL2959) (NITE BP-224), Lactobacillus bulgaricus JCM1002 strain (L. burgalicus JCM1002), and Streptococcus thermophilus ATCC19258 strain (S. thermophilus ATCC19258). First, a 10% medium of skim milk powder was used to prepare a bulk starter of Lactobacillus kawaii OLL2959 strain (NITE BP-224), Lactobacillus bulgaricus JCM1002 strain, and Streptococcus thermophilus ATCC19258 strain. Next, the Yogurt mixture (SLF-free: 9.5%, fat fraction (FAT): 3.0%) was heat-treated at 95 ° C for 5 minutes. The heat-treated yoghurt mixture was inoculated with 1% of the starting strains of Lactobacillus bulgaricus JCM1002 strain and Streptococcus thermophilus ATCC19258 strain, and 5% of the starting strain of Lactobacillus gasseri OLL2959 strain (NITE BP-224). The fermentation was carried out at 43 ° C for 4 hours to obtain the original yogurt. After the original yogurt was cooled in a refrigerator (5 ° C), the flavor and physical properties were confirmed. At this time, both flavor and physical properties were good.

(Production Example 2 of fermented milk)

The original yoghurt was prepared by Lactobacillus oryzae OLL2779 strain (Lactobacillus oris OLL2779) (NITE BP-223), Lactobacillus bulgaricus JCM1002 strain (L. burgalicus JCM1002), and Streptococcus thermophilus ATCC19258 strain (S. thermophilus ATCC19258). First, a 10% medium of skim milk powder was used to prepare a bulk starter of Lactobacillus acidophilus OLL2779 strain (NITE BP-223), Lactobacillus bulgaricus JCM1002 strain, and Streptococcus thermophilus ATCC19258 strain. Then, will be excellent The mixture (SLF: 9.5%, fat fraction (FAT): 3.0%) was heat treated at 95 ° C for 5 minutes. The heat-treated yoghurt mixture was inoculated with 1% of the starting strains of Lactobacillus bulgaricus JCM1002 strain and Streptococcus thermophilus ATCC19258 strain, and 5% of the starting strain of Lactobacillus acidophilus OLL2779 strain (NITE BP-223). Fermentation was carried out at 43 ° C for 4 hours to obtain the original yogurt. After the original yogurt was cooled in a refrigerator (5 ° C), the flavor and physical properties were confirmed. At this time, both flavor and physical properties were good, and it was confirmed that Lactobacillus sp. OLL2779 strain (NITE BP-223) was extremely suitable for food production.

Industrial availability:

The present invention provides a lactic acid bacterium which can lower the uric acid value in blood. Since the lactic acid bacteria of the present invention can reduce the uric acid value in the blood by oral ingestion, the lactic acid bacteria of the present invention can be applied to foods or medicines for preventing and/or treating gout and hyperuricemia. In particular, the lactic acid bacteria of the present invention have been confirmed to have no problem of gas production in food production, and are suitable for use in foods. Further, with the lactic acid bacteria of the present invention, it is conceivable to produce a processed food having a reduced amount of mash. Therefore, the lactic acid bacteria of the present invention are highly utilized in the food and pharmaceutical industries.

Fig. 1 is a schematic diagram showing the decomposition ability of each lactic acid bacterium when various lactic acid bacteria are cultured in the presence of sputum (inosine). A strain (asterisk) which is considered to have a high decomposition ability is used as an experimental object of the animal model.

Fig. 2 is a schematic diagram showing the decomposition ability of each lactic acid bacterium when various lactic acid bacteria are cultured in the presence of sputum (inosine). Recognized as high decomposing ability The strain (asterisk) was used as an experimental object of the animal model.

Fig. 3 is a graphical representation of the decomposing ability of Lactobacillus fermentum (L. fermentum) and Lactobacillus brevis (L. brevis) by (hypoxanthine + guanine) / 5-bromouracil.

Fig. 4 is a schematic diagram showing the results of measuring the serum uric acid value by orally administering a lactic acid bacteria (fermentative lactobacillus, short Lactobacillus) having a high decomposing ability to an animal model of esophageal hyperuricemia.

Fig. 5 is a schematic diagram showing the results of measuring the serum uric acid value by oral administration of a lactic acid bacteria (Lactobacillus brevis and Lactobacillus kawaii) having a high sputum decomposition ability to an animal model of esophageal hyperuricemia. In the Lactobacillus brevis OLL2779 strain administration group (Group 7) on the 2nd and 5th days after the start of administration, and in the Lactobacillus kawaii OLL2959 strain administration group (Group 9) after the start of administration 5th On the day, a significant inhibition of the increase in serum uric acid value (# in the figure) was observed.

Claims (9)

A Lactobacillus lactic acid bacterium, which is Lactobacillus gasseri OLL2959 (Accession No.: BCRC 910379), has a deuterium decomposition ability and has no gas producing ability. A food or drink which has an inhibitory effect on an increase in uric acid value in blood, which comprises the lactic acid bacterium as described in claim 1 and/or the lactic acid bacteria-containing substance. A pharmaceutical product which has an inhibitory effect on an increase in uric acid value in blood, which comprises the lactic acid bacterium as described in claim 1 and/or the lactic acid bacteria-containing substance. A pharmaceutical product for preventing and/or treating hyperuricemia, which comprises the lactic acid bacteria and/or the lactic acid bacteria content as described in claim 1 of the patent application. A food or drink for preventing and/or treating hyperuricemia, which comprises the lactic acid bacteria and/or the lactic acid bacteria contents as described in claim 1 of the patent application. A method for suppressing the amount of cockroaches taken up by a foodstuff, characterized by administering the lactic acid bacterium according to the first aspect of the patent application and/or the lactic acid bacteria-containing substance. A method for producing a food or drink for reducing the amount of blood in a blood, which comprises the step of contacting a raw material or an intermediate product of a food or beverage with a lactic acid bacterium as described in claim 1 and/or a lactic acid bacteria-containing substance. a lactic acid bacterium as described in claim 1 and/or The lactic acid bacteria-containing substance is used for the prevention and/or treatment of a disease or a symptom of any one or more of the group consisting of hyperuricemia, gout, renal dysfunction, urinary calculi, and arteriosclerosis Use of food or beverages or pharmaceuticals. A lactic acid bacterium according to the first aspect of the invention, and/or the lactic acid bacterium-containing material, for use in the manufacture of a food or drink and/or a pharmaceutical product having an inhibitory effect on an increase in uric acid value in blood of a subject.