JP2003081855A - Lipid metabolism-improving agent and food containing the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a lipid metabolism-improving agent capable of suppressing absorption of cholesterol from the intestinal tract and lowering the cholesterol concentration in blood and having the effect of preventing and improving arteriosclerosis. SOLUTION: This lipid metabolism-improving agent comprises a fermented soybean milk obtained by making lactic acid bacterium act on the soybean milk.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a lipid metabolism improving agent containing fermented soy milk as a main component, which is obtained by allowing lactic acid bacteria to act on soy milk, a food containing the same, and a low-density lipoprotein antioxidant.

[0002]

2. Description of the Related Art Heretofore, as a method for preventing / ameliorating arteriosclerosis, it has been generally practiced to reduce blood cholesterol level. Such a method involves hypercholesterolemia, especially high LDL (low specific gravity). Lipoprotein) Effective in improving cholesterolemia (Hiroshi Mabuchi et al. (198
7) Coronary vol4,281). On the other hand, the occurrence of arteriosclerosis is L
It is reported that the oxidative modification of DL starts.
nberg D et al. (1989) New Engl J Med, 321, 1196-1197), and a substance having an antioxidant activity against LDL is drawing attention. Further, soybean milk contains soybean protein, phospholipids, and isoflavones, and is expected to be effective in lipid metabolism. However, soy milk has a peculiar unpleasant odor and unpleasant taste, and is currently shunned by many consumers.

[0003]

SUMMARY OF THE INVENTION Therefore, an object of the present invention is to provide a lipid metabolism improving agent which is excellent in preventing and improving arteriosclerosis.

[0004]

[Means for Solving the Problems] In view of such circumstances, the present inventors have focused their attention on the active ingredient of soy milk, and as a result of earnest research, fermented soy milk obtained by allowing lactic acid bacteria to act on soy milk is The present invention has been completed by discovering that it has an excellent antioxidant activity against LDL and an effect of suppressing lipid absorption from the intestinal tract, and is useful as a lipid metabolism improving agent.

That is, the present invention comprises a fermented soymilk obtained by allowing lactic acid bacteria to act on soymilk, a lipid metabolism improving agent, an LDL antioxidant, a cholesterol intestinal absorption inhibitor, and the same. The present invention provides a food for improving lipid metabolism.

[0006] It has been attempted to ferment soy milk with lactic acid bacteria or bifidobacteria for the purpose of reducing the unpleasant odor peculiar to soy milk, but fermented soy milk still has an excellent lipid metabolism improving effect. Not reported.

[0007]

BEST MODE FOR CARRYING OUT THE INVENTION The lipid metabolism improving agent of the present invention contains fermented soymilk obtained by reacting lactic acid bacteria with soymilk as a main component. Here, the fermented soybean milk used in the present invention is obtained by allowing lactic acid bacteria to act on soybean milk, and examples thereof include those produced according to the following steps.

(1) Production of Soymilk (Step 1) The soymilk used in the present invention may be produced by any method. For example, soybean as a raw material is soaked in water, and then hot water or 0. It can be produced by adding hot water containing 5 to 1.0% by weight (hereinafter, simply indicated by%) of sodium carbonate, grinding and then removing okara, and further sterilizing by heating.

The soybean used as the raw material for soybean milk used in the present invention is not particularly limited, but it is preferable to use raw soybeans containing oils and fats, dehulled soybeans, flaked soybeans, etc., especially dehulled soybeans. It is preferable to use as a raw material.

If necessary, the soymilk may be prepared from sugar such as sucrose, glucose, fructose and invert sugar, which are used in foods, and a meat extract for the next step of microbial treatment after production. The nutrients necessary for the growth of microorganisms such as peptone, yeast extract, and peptides may be added, and the optimum pH of the microorganisms may be added.
In order to adjust the amount to the soybean milk, citric acid, malic acid, ascorbic acid, lactic acid, acetic acid and other acids used in food may be added.

(2) Fermentation of soybean milk (step 2) The method for allowing lactic acid bacteria to act on the soybean milk produced in the above step 1 is not particularly limited. For example, after inoculating the soybean milk with the cultured lactic acid bacterium solution, the microorganism Fermentation may be performed by appropriately determining conditions such as temperature, time, and anaerobic conditions suitable for anaerobic bacteria. The fermentation may be mixed fermentation in which a plurality of strains are combined, or continuous fermentation in which a plurality of strains are combined.

[0012] Here, the lactic acid bacteria acting on soybean milk, unless the lactobacillus itself or its metabolites are harmful to the human body,
Lactobacillus genus, Lactococcus genus, Streptococcus genus or lactic acid bacteria belonging to Leuconostoc genus are preferable, but not particularly limited, and particularly Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus Helveticas, Lactobacillus gasseri, Lactobacillus salivarius salivarius, Lactobacillus salivarius salicinius, Lactobacillus buchineri, Lactobacillus fermentum, Lactobacillus paracasei tolerance, Lactobacillus paracasei paracasei, Lactobacillus Sony, Lactobacillus delbrucki delbrucki, Lactobacillus delbrucki bulgaricus, Lactobacillus delbrucki lactis, Kutobachirusu-Garinaramu, Lactobacillus amylovorus, Lactobacillus brevis,
Brevis, Lactobacillus rhamnosus, Lactobacillus kefir, Lactobacillus crispatas, Lactobacillus reuteri, Lactococcus lactis
Lactis, Lactococcus lactis cremoris, Lactococcus plantarum, Lactococcus raffinolactis, Streptococcus thermophilus, Leuconostoc mesenteroides cremoris, Leuconostoc lactis and the like are preferable.

Fermented soy milk obtained by allowing lactic acid bacteria to act on soy milk can be directly used as the lipid metabolism improving agent of the present invention, but additives commonly used in foods and oral pharmaceuticals may be added. The additives used here are sugars,
Protein, lipid, vitamins, plant extract, animal extract,
Examples thereof include gelling agents, fragrances, coloring agents and the like. The fermented soymilk used for the lipid metabolism improving agent of the present invention can be used as it is or without sterilization.

The dose of the lipid metabolism-improving agent and the low-density lipoprotein antioxidant of the present invention used as a drug varies depending on the administration method, age, weight and condition of the patient. Against 10 per day
It is preferably 0 to 500 ml.

The lipid metabolism-improving agent of the present invention can be used by adding it to food in an arbitrary range to give a lipid metabolism-improving food. As food, lactic acid bacteria drink,
It may be contained in fermented milk, soy milk, milk, cheese, pudding, ice cream or the like in an amount of about 10 to 80%, preferably about 40 to 70%, and may also be contained in biscuits, bread or the like.

[0016]

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

Example 1 1% glucose (W / V) in soybean milk (made by Shikoku Kakoki)
In addition, the pH was adjusted to 6.8 with 2M citric acid and then 100
What was sterilized by steam at 30 ° C for 30 minutes was used as a soymilk medium. A seed starter of human-derived lactic acid bacteria was inoculated into this in an amount of 0.5 to 1%, and cultured at an optimum temperature (26 to 37 ° C) for 7 days.
After culturing, take 1 g of each bacterial solution into a test tube with a stopper, add 7 ml of methanol, and stir at 4 ° C overnight with occasional stirring.
I left it at. Then, the mixture was centrifuged (1500 xg, 5 ° C), and the supernatant was diluted to 10 ml to prepare a 10-fold diluted sample. This was stored at −20 ° C. until the measurement, and further diluted 4 times before use when measuring the antioxidant property.

Syrian hamsters (male 6 weeks old) were preliminarily fed with MF feed for 1 week, and then fed with MF feed supplemented with 0.5% cholesterol and 5% lard for 2 weeks. On the day before dissection, the animals were fasted for 24 hours, blood was collected from the abdominal aorta, and plasma was prepared by the EDTA method. An LDL fraction was collected from this plasma by an ultracentrifugation method, analyzed with a physiological phosphate buffer for 24 hours, and then diluted to an appropriate concentration to give an LDL for oxidation reaction.

Negative charge increases when LDL undergoes oxidative modification, and migration distance increases when electrophoresis is performed on agarose gel. Therefore, the antioxidant property of fermented soybean milk is examined using the increase in migration distance as an index of oxidation. It was

LDL (final concentration 500 μg / ml protein) is
After the addition of the sample, the mixture was incubated in the presence of 5 μM CuSO 4 for 6 hours at 37 ° C. for oxidation, and after the addition of EDTA, the reaction was stopped by cooling in ice water. Further, the one to which methanol was added instead of the sample was used as a control, and the one which was incubated at 0 ° C. after adding methanol and EDTA was used as a blank.

1 μl of the reaction solution was mixed with 1% agarose gel (multitrack LDH isozyme gel 16: CIBA-CORNIN
G), electrophoresed at 90 V for 45 minutes, and then stained with Cholecholest A (Nippon Chemifa) for cholesterol staining. After the gel was dried, the migration distance of LDL in each lane was measured. The blank was assumed to be in a state where LDL was not oxidized at all, and the rate at which the increase in the migration distance was suppressed by the addition of the sample as compared with the control was defined as the LDL denaturation suppression rate.

The calculation formula is shown in (1). The antioxidant property was expressed as a specific inhibition rate when the value of the soymilk medium was 100.

[0023]

[Equation 1]

[0024]

[Table 1]

From the above results, the fermented soybean milk produced by lactic acid bacteria had LDL antioxidant activity, and it was confirmed that the fermented soybean milk had excellent LDL antioxidant activity as compared with soybean milk.

Example 2 Lactic acid bacteria precultured in a medium containing 0.5% yeast extract and glucose as needed in 10% skim milk powder were supplemented with 1% glucose and the pH was adjusted to 6.8 to 6. The soybean milk adjusted to 9 (manufactured by Shikoku Kakoki Co., Ltd., sterilized at 100 ° C. for 90 minutes) was inoculated with 0.5 to 1% and cultured at a predetermined temperature for 2 days.

200 mg of fermented soy milk prepared as described above
In addition, 200 μm of artificial lipid micelle prepared by the method described below is used.
After adding l, leave at 37 ° C for 1 hour, then centrifuge (8000 x
g, 15 minutes) and determine the cholesterol concentration in the supernatant
It measured using 555 (Kyowa Medix). As a control, soybean milk was used, and the amount of cholesterol transferred to the precipitate of each fermented soymilk was defined as the cholesterol insolubilization rate according to the following formula (2).

[0028]

[Equation 2]

[Preparation of artificial lipid micelle] 2 g of oxgall (manufactured by DIFCO) was added to 75 ml of phosphate buffer (150 mM, pH 7.0).
Cholesterol (manufactured by Wako Pure Chemical Industries, Ltd.) 921 mg and lysophosphatidylcholine (manufactured by SIGMA) 135 mg were added in this order and dissolved, and then monooleic acid (manufactured by Tokyo Chemical Industry Co., Ltd.) 90.2 mg, oleic acid ( 702.2 mg of Wako Pure Chemical Industries, Ltd.) was added and mixed, and a phosphate buffer was added to bring the total amount to 100 ml. While stirring the solution, ultrasonic treatment (SONIFR (manufactured by BRANSON), small chip) was performed at room temperature. After stirring the emulsion and the micelle solution for a while, ultracentrifugation was performed at 100,000 × g and 25 ° C. for 16 to 18 hours. After ultracentrifugation,
Only the transparent micelle layer was collected to prepare artificial lipid micelles.

[0030]

[Table 2]

[0031]

[Table 3]

From the above results, all of the fermented soymilk by lactic acid bacteria had a stronger micelle insolubilizing effect than soymilk. In order for cholesterol to be absorbed from the small intestinal mucosa, it must be dissolved in micelles. Therefore, fermented soymilk is expected to suppress cholesterol absorption more than soymilk.

[0033]

The lipid metabolism improving agent of the present invention can be expected to suppress the absorption of cholesterol from the intestinal tract and lower the blood cholesterol concentration, and therefore can be expected to have an effect of preventing and improving arteriosclerosis. Moreover, since the lipid metabolism improving agent of the present invention has an excellent antioxidant activity against LDL, it is possible to effectively prevent the oxidative degeneration of LDL which is the origin of arteriosclerosis. Furthermore, the lipid metabolism improving agent of the present invention is fermented soy milk obtained by allowing lactic acid bacteria to act on soy milk, so it is a sensory excellent lipid metabolism improving agent with no safety problems and arteriosclerosis. It is useful for the prevention and treatment of.

   ─────────────────────────────────────────────────── ─── Continued front page    (72) Inventor, Emi Yasuda             1-1-19 Higashishimbashi, Minato-ku, Tokyo Stock market             Company Yakult Head Office (72) Inventor Norie Onodera             1-1-19 Higashishimbashi, Minato-ku, Tokyo Stock market             Company Yakult Head Office (72) Inventor Fumiho Ishikawa             1-1-19 Higashishimbashi, Minato-ku, Tokyo Stock market             Company Yakult Head Office F-term (reference) 4B018 MD58 MD86 ME04 MF13                 4B020 LB18 LC05 LG05 LK18 LP18                 4C088 AB59 AC04 CA25 MA52 NA14                       ZA45 ZA70 ZC33

Claims (4)

[Claims] 1. A lipid metabolism improving agent, which comprises fermented soy milk obtained by allowing lactic acid bacteria to act on soy milk. 2. The lipid metabolism improving agent according to claim 1, wherein the lactic acid bacterium is one or more selected from the genera Lactobacillus, Lactococcus, Streptococcus and Leuconostoc. 3. The lactic acid bacterium is Lactobacillus casei,
Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus helveticus, Lactobacillus gasseri, Lactobacillus salivarius
Salivarius, Lactobacillus salivarius salicinius, Lactobacillus buchineri, Lactobacillus
Fermentum, Lactobacillus paracasei tolerance, Lactobacillus paracasei paracasei, Lactobacillus johnsonii, Lactobacillus delbruecki delbrukki, Lactobacillus delbrukki
Bulgaricus, Lactobacillus delbrucki lactis, Lactobacillus galinarum, Lactobacillus amylobora, Lactobacillus brevis brevis, Lactobacillus rhamnosus, Lactobacillus kefir, Lactobacillus crispatas, Lactobacillus reuteri, Lactococcus Lactis lactis, Lactococcus lactis cremoris, Lactococcus plantarum, Lactococcus raffinola lactis,
The lipid metabolism improver according to claim 1 or 2, which is one or more selected from Streptococcus thermophilus, Leuconostoc mesenteroides cremoris and Leuconostoc lactis. 4. An absorption inhibitor of cholesterol from the intestinal tract, which comprises fermented soy milk obtained by allowing lactic acid bacteria to act on soy milk.