KR100392904B1 - Composition containing egg yolk antibody, and food containing it - Google Patents

Abstract

The present invention is applied to Helicobacter Philolyli antibody-containing skim yolk powder, 0.02-0.1% by weight of catechin, 0.1-1.0% by weight glycoprotein (GMP), 5-10% by weight of sugar, 5-10% by weight relative to the total weight of the composition The present invention relates to an egg antibody-containing composition for preventing Helicobacter pylori infection, and a food containing the same, comprising a% sugar alcohol, 5-20% by weight phospholipid and purified water.

Description

Egg antibody-containing composition and food containing it {COMPOSITION CONTAINING EGG YOLK ANTIBODY, AND FOOD CONTAINING IT}

The present invention relates to an egg antibody-containing composition and a food containing the same.

Helicobacter pylori is known to not only survive in the stomach under strong acidic conditions but also to be involved in the development and recurrence of peptic ulcers such as gastric ulcer and duodenal ulcer and chronic gastritis. In addition, many studies on the causal relationship with gastric cancer have been reported through epidemiological investigations or animal experiments.

In most cases, pneumatic ulcer patients have been identified as Helicobacter pylori infection, and the method of disinfecting the bacterium has been reported to reduce the recurrence rate within 10% within 2 years. It is recognized.

Currently used treatments around the world include a combination of proton pumps, which are powerful acid secreting agents used in the treatment of peptic ulcers, and two kinds of antibiotics (Criraceromycin and Amoxicillin or Metronidasol). Kind combination therapy is widely practiced as a general therapy. The bactericidal rate of these three types of combination therapy is about 80-90%, but this method uses a large amount of antibiotics, which poses problems of side effects and resistant bacteria.

In order to solve this problem, methods for preventing and disinfecting by natural products have been invented. Japanese Patent Laid-Open No. Hei 10-287585 discloses an effective bactericidal method of Helicobacter pylori by using a food containing yolk-derived antibodies specific for Helicobacter pylori.

In general, the first step in gastrointestinal infections is the attachment of pathogenic viruses or pathogens to the organ or tissue surface. It is recognized that a specific antibody produced using such an infectious agent as an antigen is effective in preventing and treating an infectious disease by binding to a corresponding pathogen and inhibiting the pathogen from adhering to digestive organ epithelial cells.

Egg-derived antibodies are also suitable for mass production and effective in actual production, but egg yolk-derived antibodies are active proteins whose activity is derived from the structure of the hypervariable region at the tip of the antibody. Protein denaturation) is very fatal considering the activity of the antibody.

In general, since proteins are known to be weak in heat, pH, etc., the yolk-derived antibody, which is a protein, is also very difficult to use under heating and acidic conditions related to direct denaturation.

Therefore, the yolk-derived antibody has low thermal stability, pH stability, solubility, dispersibility, etc., and when the antibody-containing yolk itself is applied to the liquid food, it is difficult to apply the food to the food because the separation occurs in the food form.

On the other hand, catechin, the main component of green tea, is known to have some excellent physiological activities. Among the green tea extracts, catechins (Epigallocatechin gallate, gallocatechin gallte) have an excellent physiological activity such as weakening the production of intestinal hemorrhagic E. coli O-157 H7 and vero toxin, in vivo antibacterial activity, prevention of cancer, and improvement of intestinal flora. It is reported to have.

Glycomacropeptide (GMP), a glycoprotein derived from milk protein, is a hydrophilic part of κ-casein, and many physiological activities, such as proliferation activity of intestinal lactic acid bacteria and neutralization activity of cholera toxin, have been reported. GMP has a problem in that it is difficult to be applied to food because it has a problem that the amount of solubility is low when added to food.

The present invention for solving the above problems by adding a certain ratio of green tea extract, GMP, sugars, sugar alcohol, phospholipid to the egg-derived antibody can not only increase the protective effect of gastritis, gastric ulcer and duodenal ulcer, but also heat, pH, etc. It is an object of the present invention to provide an egg antibody-containing composition having improved heat resistance, solubility and processability and suitable for application to food, and a food containing the composition.

1 is a graph of the bactericidal effect on the Helicobacter pylori of the rats of the egg antibody-containing composition (1) and egg antibody-containing composition (2), (3), (4),

2 is a graph comparing the solubility and heat resistance of the defatted yolk solution, egg antibody-containing composition (1) and egg antibody-containing composition (2),

Figure 3 is a graph showing the effect of inhibiting adhesion to Helicobacter pylori through the urease content for the samples of Table 3.

In order to achieve the above object, according to the present invention, Helicobacter pylori antibody-containing skim yolk powder, 0.02-0.1% by weight of catechin, 0.1-1.0% by weight glycoprotein (GMP) based on the total weight of the composition There is provided an egg antibody-containing composition for preventing Helicobacter pylori infection, comprising 5-10 wt% sugar, 5-10 wt% sugar alcohol, 5-20 wt% phospholipid and purified water, and a food containing the same.

Hereinafter, the present invention will be described in detail.

The antibody-containing egg yolk used in the present invention is preferably used by degreasing in powder form.

Catechin, a high-purity green tea extract, is more effective with epigallocatechin gallate (Egcg) of about 65% or more. If catechin is added more than 0.1% to the total weight of the composition, the taste is felt and affects the flavor. If it is added less than 0.02%, the catechin is preferably adjusted to 0.02 to 0.1% by weight. Can be.

In addition, GMP, which is a glycoprotein derived from a milk protein in the present invention is not particularly limited, but is separated and purified by a known method such as an ion exchange resin method or a membrane separation method, and GMP is contained in about 70% or more of the total milk protein. It is preferable to use it because the higher the purity, the better the effect. In addition, the higher the content of sialic acid is better. GMP is preferably added in an amount of 0.1 to 1.0% by weight based on the total weight of the composition. If it is less than 0.1% by weight, there is no effect of addition, and if it is more than 1.0% by weight, there is a problem in solubility.

In the present invention, sugars and sugar alcohols are not particularly limited, but it is preferable to use high fructose or sucrose as sugars, and sugar esters as sugar alcohols. Among them, it is particularly preferable to use high fructose as the sugar and sorbitol as the sugar alcohol. The amount of sugar added is preferably 5-10% by weight of the total weight of the composition, and the amount of sugar alcohol is preferably 5-10% by weight of the total weight of the composition. This is because when the sugar and the sugar alcohol are added in less than 5% by weight, there is no effect of addition, and when it exceeds 10% by weight, the sugar content becomes too high.

In addition, soybean phospholipid (lecithin), egg yolk lecithin and the like may be used, and high purity phospholipid derived from egg yolk is more preferable in flavor and processability. The addition amount is preferably 5-20% by weight based on the total weight of the composition, since less than 5% by weight becomes too thin and more than 20% by weight becomes too viscous.

For example, the egg antibody-containing composition according to the present invention is added with sugars, sugar alcohols, phospholipids, etc., using a stirrer with a rotary stir blade, and then stirred, followed by mixing water, catechin, and GMP in this order and foaming. It is preferable to mix | blend so that it may mix.

By combining glycoproteins derived from catechins and milk proteins with skim yolk containing Helicobacter pylori, the inhibitory effect on adhesion, which is the first stage of Helicobacter pylori infection, is improved, and high purity phospholipids and sugars are added to the food addition process. The heat resistance and acid resistance which are problematic become markedly improved.

In addition, since the composition according to the present invention improves heat resistance, solubility, and dispersibility when food is added, food can be prepared by adding the egg antibody-containing composition of the present invention as described above.

Examples of the food include, but are not limited to, various beverages such as dairy products, soft drinks, and health supplement foods.

In the present invention, the term "dairy product" is not particularly limited. Examples thereof include milk (fresh milk, processed milk), baby formula, yogurt, drink yogurt, ice cream, skim milk powder, mixed milk powder, and the like.

When the egg antibody-containing composition of the present invention is added to dairy products, various beverages, and health supplements, the addition amount is not particularly limited but is preferably 1% by weight or more based on the total weight of the product.

The following examples are provided to illustrate the present invention, but the present invention is not limited thereto.

(Example 1)

(1) Preparation of Antigen

Helicobacter pylori clinical isolates were incubated for 4 days at 37 ° C. in agar medium containing 7% metal fiber (10% CO ₂ , 10% air, humidification). After incubation, the resulting colonies were removed, suspended in physiological saline, formalin was added to the suspension at 0.5% (v / v), and left at room temperature for 18 hours to inactivate the cells.

Next, the cells were collected by centrifugation at 3,000 × g for 10 minutes and washed three times with physiological saline. The cells were dispersed in physiological saline, homogenized with histotron, and the absorbance was 0.8 at a wavelength of 660 nm to obtain antigen solution A.

(2) Production of immune egg yolk

1 ml of antigen was injected per egg into the laying hens (Heytreghorn). Immunization was repeated once a week, four times in total, and immunization was performed once every month for the purpose of maintaining the antibody titer.

The specific antibody titer in egg yolk was measured by aggregate titer by microtiter method. Egg yolk is separated from the eggs collected every two weeks, 1 g of yolk and 9 ml of lambda -carrageenan aqueous solution (1.5% w / v) are mixed and left at room temperature for 30 minutes, followed by centrifugation at 1,000 × g for 10 minutes. An egg yolk soluble protein fraction containing an antibody was obtained to prepare a sample solution.

The sample solution was diluted twice with physiological phosphate buffer pH 7.4 (hereinafter referred to as PBS) to obtain a sample solution.

50 microliters of each test solution and 50 microliters of antigen solution were mixed in a microplate, and left to stand at 37 ° C for 24 hours to check for coagulation. The antigen solution was obtained by diluting the antigen used for immunization with PBS 10 times (absorbance 0.8 at wavelength 660 nm). The aggregated antigenic value is expressed as the inverse of the maximum dilution factor of the sample to be aggregated.

1 kg of egg and egg yolk obtained by coagulating antigens of more than 320 eggs were homogenized with a homogenizer. Each solution was sterilized at 63 ° C. for 3 minutes, and then powder was prepared by spray drying to obtain 470 g of egg and egg yolk powder containing specific antibodies to Helicobacter pylori. The conditions of spray drying were performed at the blowing temperature of 145 degreeC, and the ventilating temperature of 85 degreeC.

An equal amount of ethanol was added to the obtained antibody-containing yolk powder for degreasing. Lipids were repeatedly degreased to be 2% or less. Defatted egg yolk powder obtained from the antigen solution A was referred to as sample powder A.

(3) Preparation of Egg Antibody-Containing Composition

25.1% purified water was added to the obtained sample powder A to prepare a comparative egg antibody-containing composition (1), and to the sample powder A, phospholipid (10%), high fructose (5%) and sorbitol (5%) were added. Purified water, catechin and GMP were combined in the following ratios and stirred until homogenized with a homogenizer, then refrigerated.

Egg antibody-containing composition Purified water Catechin GMP (2) 4 % 0.1% One % (3) 4.98% 0.02% 0.1% (4) 4.55% 0.05% 0.5%

(Test Example 1)

Antibacterial Effect of Helicobacter Philophyllum Containing Egg Antibody-Containing Compositions

The egg antibody-containing composition prepared in Example 1 was orally administered once a day for 14 days to the rats infected with Helicobacter pylori. After 14 days, the number of Helicobacter pylori cells adhered to the gastric mucosa was measured as follows.

The stomach tissues were homogenized after 8 washes with PBS pH 7.2. After inoculation into selective medium (Poremedia H. pylori isolated medium, Eiken kagaku), and incubated for 5 days at 37 ℃ by the gas pack method.

As shown in FIG. 1, the number of helicobacter pylori in the stomach tended to decrease depending on the amount of the egg antibody-containing composition administered. Among them, the egg antibody-containing composition (2) was the most effective. In the case of increased catechin and GMP content, there was a decrease in the number of helicobacter pylori in the stomach, but when it was added more than 0.1% by weight of catechin, the taste was not good, and it was not good for flavor. It became a problem.

(Test Example 2)

Machining stability test

The egg antibody-containing compositions (1) and (2) were heated at 60-72 ° C. for 5 minutes, and then the antibody activity in the powder was measured to examine the thermal stability of the powder. An acidic solution of 4.5 was added to check pH stability. The results are shown in Tables 1 and 2, respectively.

Comparison of Thermal Stability of Egg Antibody-Containing Compositions 60 ℃ 62 ℃ 64 ℃ 66 ℃ 68 ℃ 70 ℃ 72 ℃ Remaining titer of egg antibody-containing composition (1) (antibody titer: 8000) 7500 7200 6800 6400 5600 5200 4800 Remaining titer of egg antibody-containing composition (2) (antibody titer: 8000) 8000 8000 7850 7850 7500 7450 7200 * For potency measurement, ELISA method is applied.

Comparison of pH Stability of Egg Antibody-Containing Compositions pH3.2 pH3.4 pH3.6 pH3.8 pH4.0 pH4.2 pH4.5 Remaining titer of egg antibody-containing composition (1) (antibody titer: 8000) 3200 4800 5200 6000 6400 7200 7600 Remaining titer of egg antibody-containing composition (2) (antibody titer: 8000) 4000 6400 7000 7300 7500 7500 7600

(Test Example 3)

Processing characteristics

Using the egg antibody-containing compositions (1) and (2), liquid foods (yogurts) were prepared as follows, and the separation states of the yolk solution and the yogurt culture solution were compared.

ingredient Prescription 1 Prescription 2 milk 96.0% 96.0% Skim milk powder 0.5% 0.5% Egg antibody-containing composition (2) 2.0% - Egg antibody-containing composition (1) - 2.0% Purified water 1.5% 1.5%

As a result of examining the separation status of the yolk component of the prepared food, in the food of the formula 1 containing the egg antibody-containing composition according to the present invention, the separation of the yolk component was not confirmed and the stability was maintained and good dispersibility and solubility were shown.

On the other hand, Formula 2 was confirmed that the separation of the egg yolk component floating. In addition, the sensory evaluation also showed satisfactory results with the food of prescription 1. This improvement in stability and solubility has enabled the application of liquid products such as drink yogurt.

(Test Example 4)

Synergy Confirmation Test

Preparation of Helicobacter pylori was carried out in the same manner as in Example 1. Colonies of Helicobacter pylori were collected and washed with 10 mM and PBS pH 7.4. The cells collected by centrifugation at 3,000 x g for 10 minutes are suspended again in 10 mM PBS pH 7.4, about 2 x 10 ² CFU in 1 ml.

To a 24-well polystyrene plate coated with 0.2% swine gastric mucosa mucin (Sigma) solution, the comparison with the bacteria solution degreasing egg yolk powder (10mg / ml), the egg antibody-containing composition (1), (2), catechin composition and 1 ml of each GMP composition was added, incubated for 1 hour, washed with physiological saline, and the number of bacteria of Helicobacter pylori remaining on the plate was calculated by measuring the urease produced by the same bacteria. For the activity of urease, the absorbance having a wavelength of 557 nm was determined by the indophenol method.

As a control, using a skim powder obtained from a commercial egg yolk, the amount of adhesion was investigated in the same manner as above, and the measurement results when the value was 1 were shown in Table 3 and FIG. 3.

Sample Name ingredient [One] Comparative degreasing solution Defatted yolk powder (74.9%), purified water (25.1%) [2] Egg antibody-containing composition (1) Anti-degred egg yolk (74.9%), purified water (25.1%) [3] Catechin Composition Antibody degreasing (74.9%), purified water (5%), phospholipids (10%), high fructose (5%), sorbitol (5%), catechin (0.1%) [4] GMP composition Antibody degreasing (74.9%), purified water (4.1%), phospholipid (10%), high fructose (5%), sorbitol (5%), GMP (1%) [5] Egg antibody-containing composition (2) Antibody degreasing (74.9%), purified water (4%), phospholipid (10%), high fructose (5%), sorbitol (5%), catechin (0.1%), GMP (1%) [6] Egg antibody-containing composition (3) Anti-degred egg yolk (74.9%), purified water (5.1%), phospholipid (10%), high fructose (5%), sorbitol (5%) [7] Egg antibody-containing composition (4) Anti-degred egg yolk (74.9%), purified water (25%), catechin (0.1%) [8] Egg antibody-containing composition (5) Antibody degreasing (74.9%), purified water (24.1%), GMP (1%)

In the above results, compared with the comparative skim egg yolk powder solution and egg antibody-containing composition (1), (3), (4), (5), catechin composition, GMP composition, etc., the egg antibody-containing composition (2) according to the present invention Significantly improved adhesion inhibitory effect was observed in.

(Manufacture example 1)

Preparation of foods to prevent gastritis and stomach and duodenal ulcer

Gastritis, gastric and duodenal ulcer foods were prescribed as follows.

1) Yogurt to prevent gastritis, gastric ulcer and duodenal ulcer

ingredient Content (% by weight) milk 96% Skim milk powder 0.5% Egg antibody-containing composition (2) One% Purified water 2.5%

2) Mixed milk powder to prevent gastritis, gastric ulcer and duodenal ulcer

ingredient Content (% by weight) Skim milk powder 95% Vitamins and minerals 3.5% Egg antibody-containing composition (2) One% Purified water 0.5%

3) Prevention of gastritis, gastric ulcer and duodenal ulcer

ingredient Content (% by weight) Salt-free Butter 7.0% Battery condensed milk 10.0% milk 35.0% Whole milk powder 2.5% Refined sugar 4.0% 75% Brix 14.0% Emulsifying Stabilizer 0.5% Egg antibody-containing composition (2) 1.0% water 26.0% Spices moderation

According to the present invention, the yolk-derived antibody composition having excellent bactericidal effect on Helicobacter pylori, which is effective in preventing gastritis, gastric ulcer and duodenal ulcer, as well as heat resistance, acid resistance, solubility, dispersibility and processability, and containing the same You can get food.

Claims (2)

To skim yolk powder containing Helicobacter pylori antibody, 0.02-0.1% by weight of catechin, 0.1-1.0% by weight of glycoprotein (GMP), 5-10% by weight of sugar, 5-10% by weight of the total composition An egg antibody-containing composition for preventing Helicobacter pylori infection comprising a mixture of wt% sugar alcohol, 5-20 wt% phospholipid and purified water. Food containing the egg antibody containing composition of Claim 1.