KR20020021545A - FERMENTED MILK, MILK, ICECREAM USING EGG YOLK HAVING IgY, AND ITS METHOD FOR PREPARING - Google Patents

Abstract

PURPOSE: A process of preparing the titled product by inducing microorganism harmful to the human body into laying hens and then fermenting the immunoprotein enriched egg yolk with a culture solution of lactic acid bacteria is provided, thereby producing a fermented milk product, milk and ice cream capable of providing immune function against microorganism harmful to the human body by use of an antibody present in egg yolk of artificially over-immunized laying hens. CONSTITUTION: Microorganism harmful to the human body are infused into laying hens to produce immune eggs, the egg yolk is separated from the immune eggs and then added with 5 to 10% by weight of sugar, 10 to 20% by weight of high fructose and 10 to 20% by weight of fructooligosaccharide to maintain the potency of an antibody and then sterilized at 60 to 65deg.C for 2 to 10 min to produce a solution of an egg yolk. Thereafter 0.05 to 5% by weight of the solution is added with a culture solution of lactic acid bacteria and fermented at 35 to 37deg.C for 12 to 20 hr.

Description

Fermented milk, milk, ice cream and production method using yolk solution enhanced with immunoprotein {FERMENTED MILK, MILK, ICECREAM USING EGG YOLK HAVING IgY, AND ITS METHOD FOR PREPARING}

The present invention relates to fermented milk, milk, ice cream and a method for producing the same using egg yolk with enhanced protein by injecting microorganisms harmful to the human body into a laying hen.

More specifically, the present invention is to produce an immune egg by injecting microorganisms harmful to the human body to the laying hen, and to isolate the egg yolk from the immune egg, add sugar to maintain the antibody titer, sterilization to prevent contamination of general microorganisms and yeast After treating to prepare a yolk solution, the present invention relates to lactic acid bacteria, milk, ice cream and a method for producing the yolk solution.

That is, the present invention relates to fermented milk, dairy products and a method of manufacturing the same using egg yolk solution containing an immunoprotein against microorganisms that may adversely affect human health.

Microbes that affect the human body are very diverse. Among these various microorganisms, beneficial microorganisms are purely separated for industrial use, cultured in large quantities, and industrialized. Many harmful microorganisms that do not have a beneficial effect on the human body have been studied to block the harmful effects of pathogenicity. However, even if it is a harmful microorganism, it does not cause a fatal disease to the human body or it is so gentle that it does not attract the attention of scientists. At present, it remains an unexplored field and has a negative effect on the human body. have. In recent years, these microorganisms are harmful microorganisms and have a bad effect on the human body. E. coli (Escherichia coli), Salmonella enteritis (Salmonella enteritis), or Shigella dysenteriae (Shigella dysenteriae). The harmful microorganisms are mainly pathogens that cause diarrhea through food in adults and infants, but some are weak, but some have a high mortality rate.

If the laying hens have immunity against these harmful microorganisms, the antibodies are transferred to eggs and ingested in the human body, thereby protecting them from various harmful bacteria.

On the other hand, the application of eggs to fermented milk (dairy products) was mixed in 1984 by Lin and Cunningham sterilizing 47.4% egg whites, citric acid, 28.4% alkali-treated soybean oil, and 19.0% skim milk. In addition, after adding glucose (sucrose), sucrose (sucrose), xanthan gum, vanilla (vanilla) extract, etc., L. bulgaricus and Streptococcus thermophilus ( S.thermophilus) Yogurt was developed by inoculation.

In 1984, Cunningham et al. Sterilized and cooled low-fat homogeneous oil, homogenized and mixed with egg white albumin (albumin) sterilized twice at 54.4 ° C, and then mixed inoculated Lactobacillus vulgaris and Streptococcus thermophilus. 6 hours at 40 ℃ to prepare a yogurt. In 1983, Kim et al. Prepared fermented milk by adding glucose, sucrose, and lactose to the homogeneous whole solution under a heat treatment condition of 58 ° C. for 30 minutes and lowering the pH to 4.8 by adding lactic acid bacteria.

In the case of Japan, the patent notification (Bulletin number: 57-54113) discloses a technique for preparing fermented milk by dissolving dry sterile egg white, from which lysozyme of egg white is removed, and inoculating lactic acid bacteria after adding glucose. have. Domestic patents include oils (notice: 97-574) sterilized by mixing lactose, sugar and egg whites with lactose, sugar, and water, and then lactobacillus ashdophyllus, Bifidobacterium longgum, and streptoco Disclosed is a method for preparing a lactic acid bacterium fermented beverage which is inoculated and mixed with Caucasus thermophilus. In the development of fermented milk using eggs as described above, fermented milk was prepared by mixing sugar, alkali-treated soybean oil, skim milk, etc. to promote fermentation in diluents of whole egg, egg white and egg yolk, and solved physical stability such as layer separation phenomenon of fermented milk. To do this, stabilizers such as xanthan gum were used.

The present invention is to provide a fermented milk, milk, ice cream or a method for producing the same, which maintains the activity of the immunoprotein (egg yolk antibody, IgY, Immunoglobulin Y) contained in egg yolk.

The present invention has been made in order to achieve the above object is a fermented milk manufacturing method of a preferred embodiment of the present invention is to produce an immune egg by injecting harmful microorganisms to a laying hen, maintaining the antibody titer after separating egg yolk from the immune egg Adding sugar to sterilize to prevent contamination of common microorganisms and yeast to prepare yolk solution, adding the sugar-added yolk solution to the lactic acid bacteria culture medium, and stirring, and adding the yolk solution to the lactic acid bacteria culture solution To prepare a fermented milk in a low-temperature long-term culture method.

In another preferred embodiment of the present invention, the method for preparing ice cream and milk comprises the steps of producing an immune egg by injecting harmful microorganisms to a laying hen, adding sugar to maintain antibody titer after separating egg yolk from the immune egg, and general microorganisms. And preparing a yolk solution by sterilizing to prevent contamination of the yeast, and mixing the sugar-added yolk solution with a weight ratio of 1 to 5% in skim milk powder or milk and homogenizing it.

Human harmful microorganisms include Streptococcus mutans, Staphylococcus aureus, Streptococcus pyogens, Escherichia coli, Salmonella enterosis Salmonella enteritis, or Shigella dysenteriae, is a pathogen that causes diarrhea mainly through food in adults and infants.

The sugar for maintaining the antibody titer is preferably sugar, high fructose or fructooligosaccharide, and the content of the sugar is preferably 5 to 20% by weight sugar, 10 to 20% high fructose and 10 to 20% fructooligosaccharide.

The sterilization treatment of egg yolk solution is sterilized 1 to 3 times at 60 to 65 ° C. for 2 to 10 minutes, and the yolk solution added to the lactic acid bacteria culture medium is preferably 0.05 to 5% by weight.

The fermented milk is prepared by the low temperature long-term culture method of the lactic acid bacteria culture medium to which yolk solution is added. The low temperature long-term culture method is preferably incubated at a temperature of about 25 to 37 ° C. for 12 to 20 hours.

The present inventors produce immune eggs containing antibodies from a laying hen with increased titers of antibodies against various pathogens and harmful bacteria. This is to separate only the yolk portion containing the antibody from the produced immune eggs to be used in the production of fermented milk and dairy products. In addition, the present invention was completed by studying the conditions for maintaining the antibody titer by adjusting the type, ratio, and amount of the sugars added to the yolk solution in the addition of the yolk solution to the fermented milk product.

Laying Egg Immunity of Harmful Microbes

Human harmful microorganisms distributed by the Korea Biotechnology Industry Association (KTCC), for example, Streptococcus mutans, Staphylococcus aureus, Streptococcus pyogens, S E.cherichia coli, Salmonella enteritis, or Shigella dysenteriae are incubated in a suitable medium, recovered, crushed by ultrasound, lyophilized and suspended in oil. Emulsification may be made to contact the olfactory mucosa of the laying hens or immunized by injection into the chest. Second immunization was performed three weeks after the first immunization and again immunized three weeks later to promote antibody production in laying hens.

Antibody Formation in Laying Hens

After 10 days after the second sensitization, serum was isolated by collecting blood from the wing vein of the laying hens. The separated serum was recovered and serially diluted in PBS buffer (pH 7.2) to react with the antigen immobilized on the plastic well to confirm the titer of the reaction. Antigen was prepared to immunize the laying hens pathogenic microorganisms-lyophilization after ultrasonic grinding-to 10 ㎍ / ㎖ in 0.05M carbonate buffer (pH 9.6) was fixed at 100 ℃ per well for 18 hours at 4 ℃. This is done by removing the serum reacted with the antigen and then measuring the titer of the antibody by reacting the immobilized antigen with 100 μl of the yolk dilution solution and the coloring reagent in order. The minimum dilution factor of the measured reaction values (more than 1.0 absorbance and 540 nm or more) of the control reaction value (0.3 of absorbance 540 nm) was set as the minimum titer of the antibody.

Generally, the titer of an antibody designates a dilution ratio or a unit of a certain number of reaction values that an antibody reacts with an antigen. 50% of the maximum response value is designated as the minimum titer or nonspecific according to the use of the antigen-antibody reaction. At least 2-3 times the antigenic antibody response may be expressed as the minimum titer. In the present invention, the latter was designated and used.

Preparation of Egg Yolk Using Egg Yolk of Immune Egg

Egg yolk was isolated from the egg obtained from the laying hens immunized three times or more, and then yolk (sugar-free / sugar-free) solution was prepared. Scattered immune eggs were collected when the size of the egg is greater than or equal to the standard for dividing the eggs. The eggs collected were harlaned with a harlan to separate egg yolk and egg white and homogenized each. The yolk solution was stored in a temporary storage tank and dissolved by adding sugar with gentle stirring at 20 rpm. After the sugar component was completely dissolved, the yolk solution was prepared by sterilization for 1 to 3 times at 60 to 65 ° C. for 2 to 10 minutes.

Identification of antibody titers in egg yolk solution

After 10 days after the second sensitization, egg yolk was separated from eggs laid and laid in a double PBS buffer. To the suspended yolk solution, PEG 6000 corresponding to 10% of the yolk weight was added and stirred until completely saturated at room temperature. After stirring, the precipitate was recovered by centrifugation at 5,000 × g for 30 minutes, suspended again in PBS buffer, and PEG 6000 corresponding to 4% of the initial yolk weight was added. After stirring again at room temperature, the precipitate was recovered by centrifugation and recovered at 4 ° C. Prepare by suspending at. The separated antibody was recovered and serially diluted in PBS buffer (pH 7.2) and reacted with the antigen immobilized on the plastic well to confirm the titer of the reaction. Antigen was prepared to sensitize the laying hens-pathogenic microorganisms-lyophilization after ultrasonic grinding-to 10 ㎍ / ㎖ in 0.05M carbonate buffer (pH 9.6) was fixed at 100 ℃ per well for 18 hours at 4 ℃. This is done by removing the serum that reacted with the antigen and then measuring the titer of the antibody by reacting the immobilized antigen with 100 µl of the lysate and the coloring reagent in this order. The dilution factor of the measured reaction values (more than 1.0 absorbance and 540 nm or more) of the control reaction value (0.3 of absorbance 540 nm or more) was set as the minimum titer of the antibody.

Add sugar content to protect egg yolk antibody titer

The egg yolk solution with increased antibody titer was intended to confirm the effect by adding various protein protection agents to maintain the antibody titer in the situation where the titer is difficult to be preserved in acidic fermented milk conditions for a long time. The materials used were fructooligosaccharides, galacto oligosaccharides, isomaltole oligosaccharides, sugars, lactose and high fructose at 0, 5, 10, 20 and 30% and each was applied in various ratios to distribute them in the sterilization process and fermented milk. It was confirmed that antibody titers were maintained during the period.

Manufacture of dairy products

Milk and Processed Milk

Crude oil collected from farmers is kept at 4-8 ℃, and alcohol test, microbiological test and antibiotic test are carried out and the high-temperature sterilization (UHT, 135 ℃ / 3-5 seconds) is passed. To 5% mixed and passed through a homogenizer (180 bar) to be stored homogeneously. Processed milk was prepared by adding flavor or other processed ingredients to compensate for sensory shortcomings in homogeneous milk and processed milk.

Fermented milk

Fermented milk production conditions were set in order to produce fermented milk which can maintain the titer of the antibody contained in the yolk when the fermented milk is prepared using the yolk solution of the immune egg. Existing fermented milks have generally been prepared using high temperature, short term culture methods. High-temperature short-term culture is a method of producing fermented milk by incubating at 40 ° C. for 4-5 hours using S. thermophilus and L. acidophilus, which have strong acid producing ability. However, the high temperature, short-term culture is too high acid production is not suitable for the production of fermented milk with yolk added. In the present invention, by using the low-temperature, long-term culture method incubated for more than 12 to 20 hours at 35 to 37 ℃ rather than 40 ℃ to maintain the properties of the immune components of yolk yolk added to the buffer effect of the acidic components and proteins produced in the fermented milk Developed to be conserved. The production conditions for fermented milk were L. casei HY 2177 and L. acidophilus HY2743, which are low in acid production and low in volatile flavor components. The initial lactic acid content was 0.01% and the number of live lactic acid bacteria was 1 × 10 ⁶ cfu, respectively. At this time, the preparation oil composition was more than 3% of the non-solid content and the incubation time was incubated for more than 12 to 20 hours at 35 to 37 ℃ and the acid content is less than 1%. The yolk solution prepared to protect the immune components in the fermented milk was mixed and used homogeneously.

Ice cream

Sugar, pectin, and CMC were mixed and mixed with a predetermined ratio in a skim milk powder adjusted to a fat content of 0.2-3%. After the raw material was completely dissolved, the prepared yolk solution was mixed up to 1 to 5% to prepare an ice cream raw material. The temperature of the solution was adjusted to a low temperature of about 4 ℃ to protect the immune components, and after homogeneous again filled and injected into an ice cream mixer to prepare a product.

Example 1

Antibody Stability in Fermented Milk from Normal and Immune Yolk Solutions

Egg titers of eggs obtained from laying hens without sensitizing microorganisms and egg yolks from laying hens sensitized with microorganisms were separated from egg layers used in the experiments and the titers of antibodies present in each egg yolk were confirmed. The antibodies contained in the egg yolk of each egg were identified using the method described above, and the antibody titers were as follows.

Table 1. Antibody Titers of Normal and Immune Eggs

Egg yolk dilution Antibody response (absorbance 405 nm) Remarks General column Immunity 1: 100 2.0 2.0 1: 200 1.6 2.0 1: 400 1.1 2.0 General column titer 1: 800 0.6 2.0 1: 1600 0.4 1.8 1: 3200 0.2 1.5 1: 6400 0.2 1.2 Immunity titration titer

As shown in Table 1, the antibody reaction value was measured through absorbance at 405 nm, and the high absorbance value means that the amount of protein is large. Therefore, the dilute sulfur dilution factor of general egg and immune egg is more than 1: 200. An egg yolk dilution factor (1: 400) having an absorbance of 405 nm of 1.1 at 1.1 was set to an appropriate titer, and an egg yolk dilution factor (1: 6400) having an absorbance of 405 nm of 1.1 was set at an appropriate titer. Thus, it was found that the antibody titer of the immune column was greater. In the case of the immune column, when the egg yolk dilution factor is 1: 100 to 1: 800, the measured values are identical, which means that the reaction between the color reagent and Ag-Ab is completed, that is, the saturation of the reaction. .

That is, through Table 1, the antibody to the various microorganisms are formed in the general egg, but the titer of the antibody is very low, the antibody titer of the immune egg produced in the laying hens sensitizing harmful microorganisms through the three-time immunization operation It was found that the antibody titers were high.

Example 2

Inhibitory Effect of Harmful Microorganisms Using Antibody Components of Immune Eggs

In order to confirm whether the antibody component produced in egg yolk has a substantial efficacy against harmful microorganisms, it was conducted to confirm the effect of growth inhibition on the harmful microorganisms. The antibody titer was determined by measuring the antibody titer by separating egg yolk from the immune eggs obtained from the over-immunized laying hens. The antibody titer was 1: 6,400. The harmful microorganisms were cultured to 1 × 10 ⁶ cfu / ml in Nutrient broth and centrifuged to recover the microorganisms. The recovered microorganisms were diluted to the same volume in PBS buffer (pH 7.2), and the separated yolk solution was added at various concentrations up to 0.05 to 5% and reacted at 37 ° C for 2 hours. After the reaction was completed, the reaction solution was plated in Nutrient agar and counted the number of viable cells cultured for 24 hours to confirm the microbial inhibitory effect of the immune eggs.

Table 2. Growth inhibition effect of harmful microorganisms by egg yolk antibody

Egg yolk concentration (%) Harmful microbial cell count (cfu / ml) Before yolk treatment After yolk treatment 0.05 1 × 10 ⁶ 1 × 10 ⁵ 0.1 1 × 10 ⁶ 1 × 10 ⁵ 1.0 1 × 10 ⁶ 1 × 10 ⁴ 2.0 1 × 10 ⁶ 1 × 10 ⁴ 3.0 1 × 10 ⁶ 1 × 10 ⁴ 5.0 1 × 10 ⁶ 1 × 10 ⁴

Example 3

Activity of Egg Yolk Antibody in Fermented Milk According to Sugar Type and Added Amount

In the addition of yolk solution containing antibody to fermented milk, various sugars were used to ensure the hydrophobic function and the stability of the physical environment such as heat, pressure, and light. Fermented milk contains about 1% of lactic acid, which shows acidity, and its pH is about 4.5, which can affect the stability of protein. Therefore, the antibody, an immunoprotein in egg yolk, should be secured. In order to ensure the stability of the antibody contained in the yolk yolk yolk was added to the fermented milk when yolk and sugar were not added to determine the titer of the antibody as an immunoprotein. The maximum titer of the antibody contained in egg yolk was set to 1: 6400, and the titer of the antibody shown when yolk containing each sugar component was added to the fermented milk was confirmed. The yolk solution with or without the sugar component was added to the fermented milk having the characteristic of 1% acidity and pH 4.5, and the yolk solution added with the yolk solution was stored at 4 ° C. for 7 days, and then the yolk solution presented in the composition of the present invention. The titer was measured using the antibody titer method. As a result of checking the titer of the antibody by adding the yolk added with sugar to the fermented milk, fructooligosaccharide, sugar, and high fructose were excellent sugars. Table 3

Table 3. Antibody Titers in Fermented Milk According to Sugars Added in Egg Yolk

Constellation Sugar component Sugar content (%) Antibody Titer Sample Number Remarks Egg yolk - - 1: 6400 One Egg yolk added to fermented milk Sugar-free - 1: 1600 2 Control Fructooligosaccharide 0 1: 1600 3 - 10 1: 3200 4 20 1: 6400 5 30 1: 6400 6 Galactooligosaccharide 0 1: 1600 7 - 10 1: 1600 8 20 1: 1600 9 30 1: 1600 10 I M O 0 1: 1600 11 - 10 1: 1600 12 20 1: 3200 13 30 1: 3200 14 Lactose 0 1: 1600 15 - 10 1: 1600 16 20 1: 1600 17 30 1: 1600 18 Sugar 0 1: 1600 19 - 10 1: 6400 20 20 1: 6400 21 30 1: 6400 22 High fructose 0 1: 1600 23 - 10 1: 3200 24 20 1: 6400 25 30 1: 6400 26

Example 4

Activity of Egg Yolk Antibody in Fermented Milk According to Sugar Ratio and Addition

As shown in Example 2 above, the activity or titer of the antibody contained in the egg is changed depending on the sugar component, and various sugars are mixed to adjust the ratio without using the sugar component alone. The titer of the antibody which appeared in fermented milk when it mixed and used for was confirmed. On the basis of what was confirmed in Example 2 above, when the ratio was adjusted using fructooligosaccharide and high fructose prepared using sugar or sugar, the titration of the antibody in fermented milk was performed. The content of sugar components added to egg yolk was set to 10% of the yolk weight, and the amount of each sugar component was added based on 10. When the sugar content was 10%, the maximum antibody titer was shown. When sugar and fructooligosaccharide, sugar and high fructose were mixed in the same amount, the antibody titer was the same as that of 10% sugar alone. When either fructooligosaccharide and high fructose were mixed alone or in the same amount, synergistic effect of increasing egg yolk antibody titer protection ability could not be confirmed. Table 4

Table 4. Antibody titers of fermented milk according to the ratio and amount of sugar added to egg yolk

No. Sugar composition (10% standard) Antibody Titer Remarks Sugar Fructooligosaccharide High fructose One 10 0 0 1: 6400 - 2 5 5 0 1: 6400 - 3 0 10 0 1: 3200 - 4 0 5 5 1: 3200 - 5 0 0 10 1: 3200 - 6 5 0 5 1: 6400 -

Example 5

Determination of the content of yolk solution added to fermented milk

The acidic fermented milk described in the constitution of the present invention was prepared to determine the amount of egg yolk that can be applied as a food when the fermented milk to which the yolk antibody as an immune component is added. The addition amount of the yolk solution added to the fermented milk was carried out to 0.05% to 5.0 by weight (%) of the fermented milk. At this time, the titer of the yolk antibody for each unit volume was measured using the antibody titer method of the above yolk solution, and as a result, the titer of dilution factor of 1: 3200 or more, which is the minimum titer unit that can serve as an antibody, Decided. In the present embodiment, when yolk antibody is added to fermented milk, it is determined that the minimum amount of yolk solution should be 0.05% or more, so that the antibody can play a role for antigen, so the minimum amount that can be added to fermented milk was 0.05% or more (Table 5 ). 5% of the sugar components that can be used by mixing the minimum amount having the antibody titer ability was appropriate.

Table 5. Determination of the amount of yolk added to fermented milk

No. Egg yolk added amount (%) Antibody Titer Remarks One 0.005 1: 800 Low titer 2 0.05 1: 3200 Available 3 0.1 1: 3200 Available 4 0.5 1: 6400 Available 5 0.8 1: 6400 Available 6 1.0 1: 6400 Available 7 2.0 1: 12800 Available 8 3.0 1: 25600 Available 9 4.0 1: 25600 Available 10 5.0 1: 25600 Available

table. 6 Characteristics of manufactured fermented milk

Item Characteristics pH 4.5 Acidity (%) 1.0 Brix 18.0 importance 1.070

table. 7 Composition of Fermented Milk Using Immune Yolk

ingredient content(%) crude oil 75.0 Skim milk powder 3.0 Egg yolk 0.05-5.0 And juice 2.0 Glucose 6.0 High fructose 2.0 Purified water 7.0 to 11.95

Example 6

Method for manufacturing milk, processed milk and ice cream in addition to fermented milk

In addition to the fermented milk described in the above embodiment, the manufacturing method of milk, processed milk, and ice cream was described in the configuration of the invention, and prepared according to the standards of the manufactured product, and the yolk solution was added to be 5% or less.

Sample a (milk, processed milk): acidity 0.2 or less, sugar content 10 brix or less,

Process oil 10brix or more

Sample b (ice cream): acidity 0.2 or less, sugar content 10 brix or more,

3% solid free

The antibody titers were maintained the highest when the acidity and sugar concentrations were prepared with high and low acidity and sugar concentrations.

The titer of the antibody contained in the yolk sac is maintained according to the type, ratio, and amount of the sugars added to the yolk sac according to the present invention, and the properties thereof are maintained when the yolk sac is added to milk, processed milk, fermented milk and ice cream. I could see. Accordingly, according to the present invention, milk, processed milk, fermented milk, and ice cream can be prepared by using an antibody present in egg yolk of an artificially overimmunized laying hen, which can provide immune function against microorganisms harmful to human body.

Claims (20)

Iii) injecting harmful microorganisms into the laying hen to produce an immune egg; Ii) adding a sugar to maintain antibody titer after separating egg yolk from the immune column; iii) sterilizing to prevent contamination of common microorganisms and yeast to prepare yolk solution; iv) adding the sugar-added yolk solution to the lactic acid bacteria culture medium and stirring it; v) fermented milk production method comprising the step of preparing the fermented milk cultured lactic acid bacteria cultured yolk solution added to low-temperature long-term culture method. The method of claim 1, wherein the human harmful microorganism of step iii) is Streptococcus mutans, Staphylococcus aureus, Streptococcus pyogens, Escherichia A method for producing fermented milk, characterized in that it is selected from the group consisting of Escherichia coli, Salmonella enteritis, or Shigella dysenteriae. The method of claim 1, wherein the sugar is selected from sugar, high fructose or fructooligosaccharide. The method of claim 3, wherein the sugar content is 5 to 10% by weight sugar, 10 to 20% high fructose, 10 to 20% fructooligosaccharide. The method of claim 1, wherein the sterilization treatment of step iii) is sterilized 1 to 3 times at 60 to 65 ° C. for 2 to 10 minutes. The method of claim 1, wherein the yolk solution in step iv) is characterized in that the weight ratio of 0.05 to 5%. The method of claim 1, wherein the low-temperature long-term culture method in step v) is cultured for 12 hours to 20 hours at a temperature of 35 to 37 ℃. Fermented milk produced by the manufacturing method according to claim 1. Iii) injecting harmful microorganisms into the laying hen to produce an immune egg; Ii) adding a sugar to maintain antibody titer after separating egg yolk from the immune column; iii) sterilizing to prevent contamination of common microorganisms and yeast to prepare yolk solution; iv) milk production method comprising the step of homogenizing the sugar-added weight ratio of 1 to 5% yolk solution in sterilized milk. The method of claim 9, wherein the human harmful microorganism of step iii) is Streptococcus mutans, Staphylococcus aureus, Streptococcus pyogens, Escherichia Milk production method characterized in that it is selected from the group consisting of Escherichia coli, Salmonella enteritis, or Shigella dysenteriae. 10. The method of claim 9, wherein the sugar is selected from sugar, high fructose or fructooligosaccharide. The method of claim 11, wherein the sugar content is 5 to 10% by weight sugar, 10 to 20% high fructose, and 10 to 20% fructooligosaccharide. The method of claim 9, wherein the sterilization of step iii) is performed at 60 to 65 ° C. for 1 to 3 times for 2 to 10 minutes. Milk produced by the manufacturing method according to claim 9. Iii) injecting harmful microorganisms into the laying hen to produce an immune egg; Ii) adding a sugar to maintain antibody titer after separating egg yolk from the immune column; iii) sterilizing to prevent contamination of common microorganisms and yeast to prepare yolk solution; iv) ice cream manufacturing method comprising the step of mixing the weight ratio of 1 to 5% egg yolk solution with added sugar to the skim milk milk solution and homogenizing. The method of claim 15, wherein the human harmful microorganisms of step iii) are Streptococcus mutans, Staphylococcus aureus, Streptococcus pyogens, Escherichia Ice cream manufacturing method characterized in that it is selected from the group consisting of Escherichia coli, Salmonella enteritis, or Shigella dysenteriae. 16. The method of claim 15, wherein said sugar is selected from sugar, high fructose or fructooligosaccharide. 18. The method of claim 17, wherein the sugar content is 5 to 10% by weight sugar, 10 to 20% high fructose, and 10 to 20% fructooligosaccharide. 16. The ice cream manufacturing method according to claim 15, wherein the sterilization treatment of step iii) is performed 1 to 3 times at 60 to 65 DEG C for 2 to 10 minutes. Ice cream prepared by the manufacturing method according to claim 15.