JP2002326946A - Lipid metabolism ameliorator and food including the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a lipid metabolism ameliorator that has excellent blood hypercholesteremia inhibitory action and low-density lipoprotein antioxidative activity. SOLUTION: The objective lipid metabolism amelioration agent and lipid metabolism amelioration food mainly comprises the fermented soybean milk that is prepared by allowing a microorganism in Bifidobacterium genus to act on soybean milk as a main active ingredient.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a lipid metabolism improver containing fermented soymilk using a microorganism of the genus Bifidobacterium and a food.

[0002]

2. Description of the Related Art Soy milk contains soy protein, phospholipid and isoflavone, and is expected to be effective for lipid metabolism. However, at present, soymilk is avoided by many consumers because of its unique unpleasant odor and unpleasant taste. In order to reduce the unpleasant odor peculiar to soymilk, fermentation of soymilk with lactic acid bacteria and bifidobacteria has been attempted, but it has not yet been reported that fermented soymilk has an excellent lipid metabolism improving effect.

[0003]

In order to prevent or ameliorate arteriosclerosis, it is general to lower the blood cholesterol level. Is effective (Hiroshi Mabuchi et al. (1987) Coronary vol 4,281). It has also been reported that arteriosclerosis is initiated by oxidative degeneration of LDL (Steinberg, D. et al. (1989)).
New Engl J. Med., 321, 1196-1197), a substance having an antioxidant activity against LDL has attracted attention.

Accordingly, an object of the present invention is to provide an excellent lipid metabolism improving agent effective for preventing and improving arteriosclerosis.

[0005]

Means for Solving the Problems In view of such circumstances, the present inventors have focused on the active ingredient of soymilk and conducted intensive research. As a result, the microorganisms belonging to the genus Bifidobacterium, particularly bifidobacterium Breve, bifidobacterium longum, bifidobacterium infantis, bifidobacterium addressentis, bifidobacterium bifidum, bifidobacterium angularatum, bifidobacterium catenulatum, bifidobacterium catenulatum Fermented soymilk obtained by acting Fidobacterium pseudocatenulatum has an excellent blood cholesterol increase inhibitory effect and an inhibitory effect on lipid intestinal absorption, and is useful as a lipid metabolism improving agent. Heading, the present invention has been completed.

That is, the present invention relates to an agent for improving lipid metabolism containing fermented soymilk as a main component obtained by allowing a microorganism of the genus Bifidobacterium to act on soymilk, an agent for suppressing absorption of cholesterol from the intestinal tract, and a lipid metabolism containing the same. It is to provide an improved food.

[0007]

BEST MODE FOR CARRYING OUT THE INVENTION The soymilk used as a raw material in the present invention is preferably a raw material such as whole soybean, dehulled soybean, or flaked soybean containing fats and oils. Good.

[0008] Soy milk is prepared by immersing raw materials in water and then adding hot water or 0.1% soy milk.
After adding hot water containing sodium carbonate in an amount of 5 to 1.0% by weight (hereinafter simply referred to as%) and grinding, the okara is removed,
Further, it can be produced by heat sterilization, but the soymilk used in the present invention may be produced by any method.

In soy milk, sugars used in foods such as sucrose, glucose, fructose, invert sugar and the like for subsequent microbial treatment;
Nutrients required for the growth of microorganisms such as meat extract, peptone, yeast extract, and peptide may be added. Further, acids used in foods such as citric acid, malic acid, ascorbic acid, lactic acid, and acetic acid may be added to soymilk in order to adjust the pH to the optimum for microorganisms.

[0010] The lipid metabolism improving agent of the present invention contains, as a main component, fermented soymilk obtained by allowing a microorganism of the genus Bifidobacterium to act on soymilk. The microorganisms belonging to the genus Bifidobacterium that act on soy milk are not particularly limited, but include Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium infantis, Bifidobacterium addressentis. , Bifidobacterium bifidum, Bifidobacterium angularum, Bifidobacterium catenulatum,
Bifidobacterium pseudocatenulatum and the like.

The method of causing these microorganisms of the genus Bifidobacterium to act on soymilk is not particularly limited. For example, after inoculating the cultured soybean milk of the microorganism of the genus Bifidobacterium into the soybean milk, a temperature suitable for the microorganism may be used. For anaerobic bacteria, fermentation may be performed by appropriately determining conditions such as anaerobic time. The fermentation may be a mixed fermentation in which a plurality of strains are combined, or a continuous fermentation in which a plurality of strains are combined. Further, the above-mentioned mixed fermentation or continuous fermentation using a microorganism of the genus Bifidobacterium and other microorganisms may be used.

The fermented soymilk obtained by allowing a microorganism of the genus Bifidobacterium to act on soymilk can be directly used as the lipid metabolism improving agent of the present invention. May be added. Examples of additives used herein include sugars, proteins, lipids, vitamins, plant extracts, animal extracts, gelling agents, flavors, coloring agents, and the like. In the present invention, the fermented soymilk can be used after being sterilized.

The dosage of the lipid metabolism-improving agent and LDL antioxidant of the present invention varies depending on the administration method, the age, weight and condition of the patient. It is preferably 100 to 500 ml per day.

The lipid metabolism-improving agent of the present invention can be used by adding it to food in an arbitrary range, and can be used as a lipid metabolism-improving food. As food, lactic acid bacteria drinks,
10-80 for fermented milk, soy milk, milk, cheese, pudding, etc.
%, Preferably about 40 to 70%, and may also be contained in biscuits, breads and the like.

[0015]

The present invention will be described in more detail with reference to the following examples, but the present invention is not limited to these examples.

Example 1 Soybean milk (manufactured by Shikoku Kakoki Co., Ltd., solid content 12.0%, crude fat 2.48%,
Crude protein (4.71%) was steam sterilized at 100 ° C for 90 minutes.
bacterium breve YIT4065 (FERM P-15488) (Soy milk medium)
Was inoculated and cultured for 30 hours. PH of bacterial solution after completion of culture
Was 4.82 and the titratable acidity was 8.25. The composition of the freeze-dried fermented soymilk obtained by freeze-drying this was almost the same as the composition of the raw soymilk. As a control, a control mixture was prepared in which the protein of fermented soymilk was replaced with casein, the lipid was replaced with corn oil, and the rest was replaced with sucrose. Using these fermented soy milk, soy milk and control mixture, a cholesterol-free diet shown in Table 1 and a cholesterol-added diet shown in Table 2 were prepared, respectively. In Tables 1 and 2, the mixture of vitamins and salts is based on AIN-76.

[0017]

[Table 1]

[0018]

[Table 2]

5 week old male Syrian hamster (SLC)
Using 36 animals, MF (Oriental Yeast Co., Ltd.) solid food for 3 days, MF powdered food for 4 days, and after preliminary breeding,
Divide into 6 groups so that there is no difference in average weight (6
) Were bred on the above diet in individual bracket cages in an environment of room temperature 24 ± 1 ° C and humidity 55 ± 5%. Food and water were freely available. The intake was measured every 2 to 3 days, 7 days after the start of the test, 4 hours after fasting, dissected under Nembutal anesthesia, blood was collected from the abdominal aorta, plasma was separated, and lipid was analyzed.

The analysis showed that plasma lipids were total cholesterol, HD
Clinical test kit for L (high density lipoprotein) -cholesterol and triglyceride (Detamina TC555 / Kyowa Medix Co., Ltd., HDL-Cholesterol Test Wako / Wako Pure Chemical Co., Ltd., Triglyceride G Test Wako / Wako Pure Chemical Co., Ltd. )). For statistical processing, multiple comparison was performed by Tukey's test after analysis of variance (analysis of variance after log conversion when equal variance was not recognized). The results are shown in Tables 3 to 6. Values were expressed as mean and standard deviation (n = 6). The significance level was 0.05. Values with the same letter indicate no significant difference. In Tables 3 and 5, the final weight indicates the weight before fasting. The arteriosclerosis index was calculated according to the formula (1).

[0021]

(Equation 1)

[0022]

[Table 3]

[0023]

[Table 4]

[0024]

[Table 5]

[0025]

[Table 6]

In the cholesterol-free diet, no effects of soymilk and fermented soymilk on the final body weight, weight gain, and feed efficiency were observed. The intake amount of soymilk and fermented soymilk tended to be higher than that of the control, but no difference was detected by multiple comparison. The levels of plasma triglycerides in soymilk and fermented soymilk were halved compared to the control. Although there was no difference in total cholesterol among the three groups, HDL-cholesterol was increased in the fermented soymilk group compared to the control group, and (VLDL (ultra low density lipoprotein) + LDL) -cholesterol levels were reduced. The arteriosclerosis index was also lower in the fermented soymilk group than in the control group. In the soy milk group
HDL-cholesterol, (VLDL + LDL) -cholesterol,
Neither control nor fermented soymilk showed any difference in arteriosclerosis index.

In the cholesterol-added diet, the body weight gain, the amount of food consumed, and the feed efficiency of the soymilk and fermented soymilk groups respectively increased as compared with the control group. The plasma triglyceride level was reduced to about 1/4 in the soymilk and fermented soymilk groups as compared to the control group.
Also, the total cholesterol in the soymilk group and the fermented soymilk group was reduced by about 15% as compared with the control group. In the soymilk and fermented soymilk groups, an increase in HDL-cholesterol and a decrease in (VLDL + LDL) -cholesterol were observed, and the atherosclerosis index decreased by about half, approaching that of the control group without cholesterol.

From the above results, the fermented soy milk was clearly more evident than the control, with or without cholesterol.
An increase in HDL-cholesterol and a decrease in (VLDL + LDL) -cholesterol were observed, confirming the effect of improving lipid metabolism. Furthermore, fermented soy milk, in the absence of cholesterol,
It was confirmed that it had an effect of improving lipid metabolism clearly superior to soymilk.

Example 2 Human-derived Bifidobacterium microorganisms (10 strains) precultured in an anaerobic GAM medium were subjected to soybean milk (sterilized at 100 ° C. for 90 minutes).
Was inoculated at 1% and cultured at 37 ° C. for 48 hours. 3 in 1 ml of culture
After adding and stirring methanol at room temperature overnight at 4 ° C., the mixture was centrifuged (3000 rpm, 10 minutes) to obtain a supernatant (4-fold dilution). This was further diluted with methanol to prepare a 40-fold diluted sample.

Syrian hamster (male, 6 weeks old) was MF
After pre-breeding on a feed for one week, they were reared on an MF feed supplemented with 0.5% cholesterol and 5% lard for two weeks. One day before the dissection, the animals were fasted for 24 hours, blood was collected from the abdominal aorta, and plasma was prepared by the EDTA method. The LDL fraction was collected from this plasma by ultracentrifugation, and was treated with physiological phosphate buffer.
After dialysis for an hour, dilute to an appropriate concentration and LD for oxidation reaction
L

LDL (final concentration 250 μg / ml protein) was oxidized by adding the sample, incubating at 37 ° C. for 4 hours in the presence of 5 μM CuSO ₄ , adding EDTA, cooling to stop the reaction, and adding Thiobarbituric acid in the reaction solution. acid reactive s
The ubstance (hereinafter referred to as TBARS) was determined from the absorbance measurement by a colorimetric method. The control was prepared by adding methanol instead of the sample, and the LDL was calculated by equation (2).
The lipid oxidation inhibition rate (%) was determined. Table 7 shows the results. The antioxidant property was expressed as a specific inhibition rate when the value of sterilized soybean milk was set to 100.

[0032]

(Equation 2)

[0033]

[Table 7]

From the above results, the fermented soymilk produced by the microorganism of the genus Bifidobacterium has LDL antioxidant activity,
Excellent LDL antioxidant activity was observed as compared to soy milk.

Example 3 Bifidobacterium microorganisms (24 strains) pre-cultured in an anaerobic GAM medium were inoculated at 1% into soybean milk (manufactured by Shikoku Koki Co., sterilized at 100 ° C for 90 minutes) and incubated at 37 ° C for a predetermined time. After air culture,
Stored at -20 ° C until measurement. To 200 mg of the fermented soymilk prepared as described above, 200 μl of the artificial lipid micelle prepared by the method described below was added, and the mixture was allowed to stand at 37 ° C. for 1 hour.
00 rpm, 15 minutes) and determine the cholesterol concentration of the supernatant
It was measured using TC555 (Kyowa Medix). Raw soymilk was used as a control, and the amount of cholesterol transferred to the precipitate of each fermented soymilk was defined as the cholesterol insolubilization rate. Table 8 shows the results.

[Preparation of artificial lipid micelle] 2 g of oxgall (DIFCO), 921 mg of cholesterol (manufactured by Wako Pure Chemical Industries, Ltd.), 921 mg of lysophosphatidylcholine (SIGMA) in 75 ml of phosphate buffer (150 mM, pH 7.0) Added and dissolved in the order of 135 mg, then 90.2 mg of monooleic acid (manufactured by Tokyo Chemical Industry Co., Ltd.)
702.2 mg of oil of oleic acid (manufactured by Wako Pure Chemical Industries, Ltd.) was added and mixed, and a phosphate buffer was added to make a total volume of 100 ml.
While the solution was stirred, ultrasonic treatment (SONIFR, small chip) was performed at room temperature. After stirring this emulsion and micelle solution for a while, 100,000 × g (RP50-2,40,000 rpm), 2
Ultracentrifugation was performed at 5 ° C. for 16-18 hours. After ultracentrifugation, only the transparent micelle layer was collected to prepare artificial lipid micelles.

[0037]

[Table 8]

From the above results, all the fermented soymilk produced by the microorganism of the genus Bifidobacterium had a stronger micelle insolubilizing effect than the soymilk. In order for cholesterol to be absorbed from the small intestinal mucosa, it must be dissolved in micelles.
Therefore, it was suggested that fermented soymilk suppressed absorption of cholesterol more than soymilk.

[0039]

Industrial Applicability The lipid metabolism improving agent of the present invention increases HDL-cholesterol and decreases (VLDL + LDL) -cholesterol, so that an effect of preventing or improving arteriosclerosis can be expected. In particular, under the condition of no cholesterol addition, there is no difference between the control mixture and soy milk,
The lipid metabolism-improving agent of the present invention has an effect of significantly reducing (VLDL + LDL) -cholesterol, and can be expected to effectively prevent arteriosclerosis even in ordinary people. Further, since the lipid metabolism improving agent of the present invention has excellent antioxidant activity against LDL, it is possible to effectively prevent oxidative denaturation of LDL, which is a starting point of arteriosclerosis. Furthermore, since the lipid metabolism improver of the present invention is composed of fermented soy milk obtained by allowing a microorganism of the genus Bifidobacterium to act on soy milk, a lipid metabolism improver that is functionally excellent without any problem in safety. And is useful for prevention and treatment of arteriosclerosis.

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI theme coat ゛ (Reference) A61P 9/10 101 A61P 9/10 101 43/00 111 43/00 111 (72) Inventor Satoshi Matsubara Tokyo 1-1-19 Higashi-Shimbashi, Minato-ku Yakult Honsha Co., Ltd. (72) Inventor Yasuhisa Shimakawa 1-1-1 Higashi-Shimbashi, Minato-ku, Tokyo Yakult Honsha Co., Ltd. (72) Inventor Emi Yasuda Tokyo 1-1-1 Higashi-Shimbashi, Minato-ku Yakult Honsha Co., Ltd. (72) Inventor Fumibo Ishikawa 1-1-1 Higashi-Shimbashi, Minato-ku, Tokyo F-term F-term (reference) 4B018 MD58 MD87 ME04 MF13 4B020 LB18 LC05 LG05 LK18 LP18 4C088 AB61 AC04 BA08 CA03 CA25 MA17 MA52 NA14 ZA45 ZC02 ZC33

Claims (4)

[Claims] 1. A lipid metabolism improver containing fermented soymilk as a main component, which is obtained by allowing a microorganism of the genus Bifidobacterium to act on soymilk. 2. The microorganism of the genus Bifidobacterium is Bifidobacterium breve or Bifidobacterium breve.
Longum, Bifidobacterium infantis,
One or more selected from Bifidobacterium addressentis, Bifidobacterium bifidum, Bifidobacterium angularum, Bifidobacterium catenulatum and Bifidobacterium pseudocatenulatum The lipid metabolism improving agent according to claim 1. 3. An agent for suppressing absorption of cholesterol from the intestinal tract containing fermented soymilk obtained by allowing a microorganism of the genus Bifidobacterium to act on soymilk. 4. A lipid metabolism-improved food containing fermented soymilk obtained by allowing a microorganism of the genus Bifidobacterium to act on soymilk.