KR102508306B1 - Method for Producing Egg Having Improved Lutein and Zeaxanthin Content, Egg Produced By Same, and Method for Preparing Egg Yolk Oil or Egg Yolk Powder - Google Patents

Abstract

The present invention relates to a method for producing eggs with increased contents of lutein and zeaxanthin, eggs produced therefrom, and a method for obtaining egg yolk oil or egg yolk powder from the eggs, and production of eggs with increased contents of lutein and zeaxanthin according to the present invention The method can produce eggs with 2.5 to 4 times higher content of lutein and zeaxanthin than normal eggs, and the color and taste of the prepared eggs are excellent, so that consumers' preference is high and health functional food that is beneficial to health can be provided. . In addition, according to the present invention, by preparing egg yolk oil and egg yolk powder having a high content of lutein and zeaxanthin from the egg, it can be applied to various functional foods that are beneficial to eye health, particularly capable of improving macular degeneration, as well as storage stability and taste. This excellent food can be provided.

Description

Method for producing eggs with increased lutein and zeaxanthin content, eggs produced therefrom, and method for producing egg yolk oil or egg yolk powder from the eggs Preparing Egg Yolk Oil or Egg Yolk Powder}

The present invention relates to a method for producing eggs with increased contents of lutein and zeaxanthin, eggs produced thereby, and a method for producing egg yolk oil or egg yolk powder from the eggs, and more particularly, to microalgae containing large amounts of lutein and zeaxanthin. It relates to a method for producing eggs with greatly increased lutein and zeaxanthin contents by feeding the eggs to laying hens, and producing egg yolk oil or egg yolk powder with increased lutein and zeaxanthin contents from the eggs produced.

Macular degeneration is a disease that causes visual impairment due to degeneration of the macula, a nerve tissue located in the center of the inner retina of the eye. Since most of the photoreceptor cells are gathered in the macula and the place where the image of an object is formed is also the center of the macula, the macula is the center of vision. plays a very important role in

Macular pigment reduces age-related macular deterioration caused by the central part of the retina and plays a role in preventing damage to retinal tissue caused by bright light. (xanthophyll). Pigments belonging to xanthophylls include lutein and zeaxanthin.

Lutein acts as an antioxidant that protects the inside of the eye from damage by oxygen free radicals produced naturally in the body, kills cancer cells by reducing the growth of blood vessels that supply cancerous tumors, and kills cancer cells in the breast, colon, and lungs. , It is known to have some effect on preventing ovarian cancer and skin cancer.

Animals cannot produce xanthophylls, and can only be obtained through food intake. Such xanthophylls exist together with chlorophyll and carotene in green parts of plants, such as leaves, flowers, and fruits. The recommended daily intake of lutein/zeaxanthin is 10mg/day, which is difficult to meet with normal dietary intake.

Lutein and zeaxanthin are usually extracted from marigold flowers, but marigold has a problem of low production compared to the cultivation area, long cultivation period, and high unit price due to the large number of resources used for cultivation.

In order to solve these problems, research for producing lutein or zeaxanthin from microalgae has been continued. For example, Korean Patent Registration No. 10-2019448 describes a method for producing lutein from Chlorella HS3, and Korean Patent Registration No. 10-2020144 uses Oxenochlorella protothecoide MM0011 strain to obtain a high concentration of lutein. describes how to produce it.

However, studies on obtaining xanthophyll pigments from microalgae have been difficult to obtain at least a certain amount of both lutein and zeaxanthin because the content of zeaxanthin is too low.

In addition, a study on a method of obtaining eggs in which macular pigment is accumulated by feeding laying hens, rather than directly extracting macular pigment from microalgae, is described in Korean Patent Publication No. 10-2017-0121052.

However, methods for obtaining eggs having high lutein and zeaxanthin contents have not yet been optimized, and studies on utilizing the produced eggs as food compositions are insufficient.

An object of the present invention is to provide a method for producing eggs with increased lutein and zeaxanthin content by feeding microalgae.

Another object of the present invention is to provide an egg produced by the above method.

Another object of the present invention is to provide a method for extracting egg yolk oil from eggs in which the lutein and zeaxanthin are increased.

Another object of the present invention is to provide a method for preparing egg yolk powder from eggs enriched in lutein and zeaxanthin.

Another object of the present invention is to provide a food composition comprising the egg yolk powder.

In order to achieve the above object, the present invention includes the step of obtaining eggs by feeding a feed or feed additive containing 1.5 to 5% by weight of microalgae containing lutein and zeaxanthin to laying hens, wherein the content of lutein and zeaxanthin is It provides a method for producing an enlarged egg.

In the present invention, the microalgae are preferably Dunaliella sp., Chlamydomonas sp., or genetic mutants thereof.

In the present invention, the microalgae is preferably included in 1.5 to 3.0% by weight of the total feed or feed additives.

In the present invention, the feed or feed additive is preferably fed for 7 to 60 days.

The present invention also provides an egg with increased lutein and zeaxanthin contents produced by the above method.

The present invention also includes (a) separating the egg yolk from the egg; (b) adding an organic solvent to the egg yolk and stirring; (c) separating the supernatant by centrifuging the mixture; and (d) evaporating the solvent from the supernatant to obtain egg yolk oil.

In the present invention, the organic solvent is preferably ethanol.

In the present invention, after step (c), it is preferable to repeat steps (b) and (c) until the color of the precipitate becomes colorless.

The present invention also provides egg yolk oil with increased contents of lutein and zeaxanthin, prepared by the above method.

The present invention also includes (a) separating the egg yolk from the egg; (b) homogenizing the egg yolk; and (c) gelatin, lactose, glucose, starch, modified starch, starch digest, gum arabic, tragant gum, dextrin, maltodextrin, collagen, casein, egg white albumin, sodium alginate, skim milk powder, whole milk, soybean protein, After mixing with one or more excipients selected from the group consisting of soy milk, cacao oil, coconut oil, animal oil, plant extract, methyl cellulose and carboxymethyl cellulose, spray drying while maintaining the product temperature at 35 to 45 ° C. It provides a method for producing egg yolk powder with increased lutein and zeaxanthin content.

In the present invention, the excipient is preferably a mixture of gelatin, lactose, pre-starch phosphate, gum arabic and maltodextrin.

In the present invention, the excipient comprises 20 to 25% by weight of gelatin, 15 to 20% by weight of lactose, 15 to 20% by weight of prestarch phosphate, 15 to 20% by weight of gum arabic, and 15 to 20% by weight of maltodextrin. desirable.

The present invention also provides an egg yolk powder with increased contents of lutein and zeaxanthin, prepared by the above method.

The present invention also provides a food composition comprising the egg yolk powder.

In the present invention, the food composition is preferably a mayonnaise composition.

The method for producing eggs with increased contents of lutein and zeaxanthin according to the present invention can produce eggs with 2.5 to 4 times higher content of lutein and zeaxanthin than ordinary eggs, and the produced eggs have excellent color and taste, which is good for consumers. It is possible to provide a health functional food that is beneficial to health while having a high degree of preference.

In addition, according to the present invention, by preparing egg yolk oil and egg yolk powder having a high content of lutein and zeaxanthin from the egg, it can be applied to various functional foods that are beneficial to eye health, particularly capable of improving macular degeneration, as well as storage stability and taste. This excellent food can be provided.

1 shows a process flow chart of the extraction method of egg yolk oil of the present invention.
Figure 2 compares the egg yolks of eggs produced from laying hens fed a regular feed and a feed containing 0.8% by weight of Dunaliella tertiorecta mutant.
Figure 3 shows the results of measuring the chromaticity of the egg yolks of the control group and the experimental group fed with 0.2, 0.4 and 0.8% by weight of microalgae.
Figure 4 shows the results of measuring the lutein and zeaxanthin contents of egg yolks produced according to an embodiment of the present invention.
Figure 5 shows the results of calculating the amount of pigment increase in egg yolk for the contents of fed lutein and zeaxanthin (lutein: 6.6mg/g, zeaxanthin: 3.5mg/g).
6 shows egg yolk oil prepared according to an embodiment of the present invention.
Figure 7 shows a comparison between the egg yolk oil emulsion of the present invention and a commercially available egg yolk oil emulsion.
8 shows egg yolk powder prepared according to an embodiment of the present invention (A: egg yolk powder prepared by spray drying commercially available egg yolks, F: egg yolk powder prepared by freeze-drying commercially available egg yolks, eB: lutein and Egg yolk powder prepared by spray-drying zeaxanthin-enriched egg yolk using three excipients, eC: Egg yolk powder prepared by spray-drying lutein and zeaxanthin-enriched egg yolks of the present invention using 5 excipients, eF: Egg yolk powder prepared by freeze-drying egg yolk enriched with lutein and zeaxanthin).
9 shows mayonnaise prepared according to an embodiment of the present invention.
10 shows the results of storing 5 types of mayonnaise for 3 weeks.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In general, the nomenclature used herein is one well known and commonly used in the art.

The present invention relates to a method for producing eggs with increased lutein and zeaxanthin content.

The present invention provides a method for producing eggs with significantly increased lutein and zeaxanthin content by feeding microalgae with increased macular pigment content, particularly lutein and zeaxanthin, to laying hens.

As the microalgae, it is preferable to use microalgae containing a large amount of macular pigment, particularly lutein and zeaxanthin. For example, Dunaliella sp., Chlamydomonas genus It is more preferable to use microalgae such as sp.), and Dunaliella tertiolecta and Chlamydomonas reinhardtii are most preferable.

The Dunaliella genus algae are unicellular green algae with isotonic biflagellates, cells are spindle-shaped, the size is 7 to 25 μm in length depending on the type, and has an embryonic chloroplast. It can grow in an environment with high salt concentration. 67% and 27% of the dry weight have the ability to be produced as lipids and carbohydrates, respectively, and maintain a pure culture by showing a rapid growth rate even in a high-salinity environment, and there is no hard cell wall, so the product is It has the advantage of being easy to extract and genetically manipulate.

In addition, the Chlamydomonas reinhardtii ( Chlamydomonas reinhardtii ) is a eukaryotic organism distributed in various environments such as freshwater and marine, and has a doubling time of 6 to 8 hours. In addition, as one of the most widespread microalgae model systems, it can be produced in a bioreactor.

In a preferred embodiment of the present invention, as the microalgae, macular pigments, that is, microalgae having improved contents of lutein and zeaxanthin may be used through genetic mutation.

In the present invention, "mutation" refers to a change in nucleotide sequence due to insertion, deletion, or substitution of bases in the original nucleotide sequence. It means mutation, and the number of bases to be inserted may vary depending on the mutation, and is not limited accordingly. "Delete mutation" refers to a mutation in which a base is lost in an original nucleotide sequence, and "substitution mutation" refers to a change in an original nucleotide sequence to another base without a change in number.

Microalgae mutants usable in the present invention, for example, Dunaliella sp. MP3, KCTC 12990BP) algae described in Republic of Korea Patent Registration No. 10-1901608 can be used. there is. The mutant strain has a significantly higher zeaxanthin production capacity per cell than conventional wild-type Dunaliella genus algae, and the original alga Dunaliella sp. Zea1, BIOTECHNOLOGY AND BIOENGINEERING , VOL. 81, NO.1) Compared to zeaxanthin, the accumulation of zeaxanthin per cell is 20 to 30% or more, so it can be effectively used as an alga for xanthophyll production.

The microalgae may be supplied to laying hens in the form of feed or feed additives, and may be preferably mixed with general feed. It is preferable to use the microalgae of the present invention by centrifuging and concentrating them, then mixing them with feed in a state in which moisture is removed as much as possible, or used in a dried state by completely removing moisture using an oven.

In the present invention, in order to produce eggs with increased lutein and zeaxanthin content, the feed fed to laying hens may contain 0.1 to 10% by weight of the total feed amount of the microalgae, preferably 1.5 to 5% by weight. It can be used, and most preferably 2 to 3% by weight.

In one embodiment of the present invention, it was confirmed that the macular pigment content of eggs gradually increased as the amount of feed added to microalgae increased, but did not increase or rather decreased after a certain point. Therefore, it was confirmed that eggs having the highest lutein and zeaxanthin contents could be produced in the range of 2 to 3% by weight of the feed of microalgae.

However, since it is not efficient to add a large amount of microalgae mutants that are difficult to obtain or expensive, it is preferable from another aspect to feed within an optimal range of pigment increase compared to the amount added. In one embodiment of the present invention, as a result of examining the amount of pigment increase in eggs with respect to the amount of lutein or zeaxanthin added in the feed, the highest amount of pigment increase was shown at the feeding rate of 0.8% by weight, and as the feeding rate increased, it was confirmed that the efficiency gradually decreased. . Therefore, when considering the increasing efficiency of macular pigment, it can be fed within the range of 0.5 to 3.0% by weight, preferably 0.6 to 2.5% by weight, and most preferably 0.6 to 1.2% by weight.

Therefore, when considering both the content of lutein and zeaxanthin and the efficiency of pigment increase compared to the added amount, the highest lutein and zeaxanthin content can be obtained in the range of 1.5 to 3.0% by weight, and efficiency can be obtained, and 1.5 to 2.5% by weight is most preferable

In the egg production method of the present invention, the feed for laying hens can be supplied once or several times daily for 7 to 60 days, preferably for about 2 weeks. In addition, obtaining eggs 7 days after the start of spawning is suitable in terms of stable egg production yield after the start of spawning.

In one embodiment of the present invention, it was confirmed that the eggs produced by the method of the present invention had about 3.8 times the lutein content and about 2.6 times the zeaxanthin content compared to eggs produced by feeding normal feed.

The present invention also relates to a method for producing egg yolk oil using eggs with increased lutein and zeaxanthin contents produced by the above method.

In the present invention, "egg yolk oil" is a component extracted from the yolk of an egg, and contains a large amount of lecithin, which is very beneficial to the human body, and contains phospholipids, which are components of cell membranes, as well as vitamins A and E, linoleic acid, etc. It is characterized by containing the macular pigments lutein and zeaxanthin.

In general, methods for producing egg yolk oil include a socho method of heating and pressing, a supercritical extraction method, and a solvent extraction method using an organic solvent. The socho method has the advantage that the process is simple and the cost is low, but it has the disadvantage that the functional component of the egg is destroyed and the consumer preference is very low due to the heat smell because it is treated at high temperature.

In the present invention, egg yolk oil is prepared using a solvent extraction method that can be treated at room temperature and is easy to preserve physiologically active substances. In particular, a method for extracting egg yolk oil using an organic solvent harmless to the human body is provided.

1 shows a process flow chart of the extraction method of egg yolk oil of the present invention.

The method for producing egg yolk oil with increased macular pigment content according to the present invention includes the steps of (a) separating egg yolk from eggs; (b) adding an organic solvent to the egg yolk and stirring; (c) separating the supernatant by centrifuging the mixture; and (d) evaporating the solvent from the supernatant to obtain egg yolk oil.

The organic solvent is preferably alcohol, and most preferably ethanol, which is an organic solvent safe for the human body. Solvent extraction using ethanol at room temperature can minimize the decomposition of lutein and zeaxanthin, and as a result, clear yellow oil without heating odor can be obtained.

The organic solvent is preferably added in an amount of 5 to 40 times the weight of the egg yolk, and after adding the solvent, stirring may be performed at 100 to 3,000 rpm for 1 to 60 minutes.

In the present invention, after step (c), the supernatant may be collected by repeating steps (b) and (c) until the color of the precipitate becomes colorless.

Evaporation of the solvent from the pooled supernatant can yield egg yolk oil. Evaporation of the solvent may be performed using an evaporation method commonly used in the art, for example, vacuum distillation is preferable.

According to the method of the present invention, it is possible to prepare egg yolk oil containing a high content of lutein and zeaxanthin, which are macular pigments, without being damaged, so that it can be used as a food or beverage for promoting eye health.

In addition, while maintaining the yellow color of the egg yolk, it is possible to produce egg yolk oil, which is more yellow in color than normal egg yolk, to provide food that meets consumer preference, and has a high phospholipid content of 32.3% when manufacturing beverages using egg yolk oil. Even if an emulsifier is not additionally added or added in a small amount, it is economical because an emulsion can be prepared.

It is safe to store the egg yolk oil at a temperature of 45 ° C or lower, and at a temperature of 50 ° C or higher, the content of lutein and zeaxanthin may be greatly reduced. In addition, storage in a dark environment without light can be stored for a long time without reducing the lutein and zeaxanthin content.

In the present invention, the egg yolk oil is preferably prepared in the form of an emulsion in that it is easy to use in food compositions and improves storage stability.

The egg yolk oil emulsion of the present invention may be prepared by mixing the egg yolk oil and water in a weight ratio of 1:1 to 1:10, preferably 1:3 to 1:5, and then stirring. The stirring may be performed at 5,000 to 30,000 rpm using a general stirring device such as a homogenizer, more preferably at 10,000 to 15,000 rpm, and preferably for 1 to 5 minutes.

The egg yolk oil emulsion according to the present invention has excellent stability according to temperature and salt concentration, so that long-term stability can be ensured when applied to food compositions, so it is highly utilized. The egg yolk oil emulsion of the present invention has excellent stability even when stored at room temperature, and is preferably stored at a temperature of less than 30 ° C, particularly less than 37 ° C. In addition, it is preferable in terms of storage stability of lutein and zeaxanthin to store under air exposure conditions rather than nitrogen substitution in a dark environment without light.

The present invention also relates to a method for producing egg yolk powder using eggs produced by the above method.

Egg yolk powder is powdered by drying egg yolk, and is used as a food ingredient for health functions. Conventional methods for producing egg yolk powder mainly use spray drying, but since drying is performed under high temperature conditions, useful components of eggs may be denatured and physiologically active substances may be destroyed.

According to the method of the present invention, by using a wall material having a specific component ratio, spray drying can be performed at a low temperature of about 40 ° C to proceed with powdering, and at the same time, the content of lutein and zeaxanthin is increased and the sensory quality is improved. Egg yolk powder can be prepared.

Excipients usable in the present invention are gelatin, lactose, glucose, starch, modified starch, starch decomposition product, gum arabic, tragant gum, dextrin, maltodextrin, collagen, casein, egg white albumin, sodium alginate, skim milk powder, whole milk, soybean protein , Soymilk, vegetable oil (cocoa oil, coconut oil, etc.), animal oil (fish oil, etc.), plant extract, methylcellulose, and carboxymethylcellulose may be used. In addition, the modified starch includes distarch phosphate, monostarch phosphate, phosphated distarch phosphate, acetylated distarch adipate, and acetylated distarch phosphate. phosphate), starch sodium octenyl succinate, starch acetate, hydroxypropyl distarch phosphate, and the like can be used.

Preferably, the excipient is preferably one or more selected from the group consisting of gelatin, lactose, pre-starch phosphate, gum arabic, and maltodextrin, and most preferably, a mixture of all five types is used.

In the excipients, gelatin is preferably used in an amount of 20 to 25% by weight based on the weight of the total excipient mixture, and lactose, prestarch phosphate, gum arabic, and maltodextrin are used in an amount of 15 to 20% by weight based on the weight of the total excipient mixture it is desirable to be When mixed in the above range, egg yolk powder having a high content of lutein and zeaxanthin and excellent taste can be prepared.

In the present invention, for the spray drying of egg yolk, first, the egg yolk may be homogenized at 55 to 65° C. for 1 to 20 minutes. At this time, homogenization may be performed at 5,000 to 50,000 rpm, and 20,000 to 30,000 rpm is preferable.

The homogenized egg yolk may be sterilized in a constant temperature water bath at 55 to 65° C. for 10 to 20 minutes.

The sterilized egg yolk is preferably mixed with an excipient and purified water, and spray drying may be performed while maintaining the product temperature at 35 to 45° C. in a spray drying apparatus. The spray drying may be performed in a commonly used spray drying apparatus.

The excipient may be added in an amount of 5 to 50 parts by weight, more preferably 7 to 15 parts by weight, based on 100 parts by weight of the egg yolk.

The purified water may be added in an amount of 10 to 100 parts by weight, more preferably 30 to 70 parts by weight, based on 100 parts by weight of the egg yolk.

The egg yolk powder prepared by the method of the present invention exhibits a darker yellow color than general egg yolk powder, and thus may have high consumer preference.

Egg yolk oil, egg yolk oil emulsion, or egg yolk powder having an increased content of lutein and zeaxanthin of the present invention can be effectively used as a raw material for food, health functional food, feed, or pharmaceuticals for eye health, particularly for improving macular degeneration. .

In addition, since the composition may be added to food or feed to achieve a special purpose, it may be a food composition, a food additive composition, a feed composition, or a composition for feed additives in this respect. When the composition is used for feed or food, the body's health can be maintained or enhanced by accumulated pigments and lipids, including lutein and zeaxanthin.

In the present invention, "for additives" includes all components added to food or feed other than the main raw material, and specific examples include effective active substances having functionality in food or feed or food and drug safety added for coloring, preservation, etc. in processed foods. It may be a food additive defined by the government. The food may be a health functional food. More specifically, it may be a health functional food for eye health.

In a preferred embodiment of the present invention, the food composition may be a mayonnaise composition. In one embodiment of the present invention, when mayonnaise was prepared using egg yolk powder with increased lutein and zeaxanthin contents, it was confirmed that consumer preference was the best in terms of color, taste, aroma, stickiness, and flavor.

In the present invention, the feed composition may be prepared in the form of fermented feed, formulated feed, pellet form, and silage.

The composition is mixed with carriers and flavorings commonly used in the food or pharmaceutical field to form tablets, troches, capsules, elixirs, syrups, powders , it can be prepared and administered in the form of a suspension or granule. As the carrier, binders, lubricants, disintegrants, excipients, solubilizers, dispersants, stabilizers, suspending agents, and the like may be used. The administration method may be oral, parenteral or application method, but oral administration is preferred. In addition, the administration dose can be appropriately selected depending on the degree of absorption, inactivity rate and excretion rate of the active ingredient in the body, age, sex, condition of the recipient, and the like. The pH of the composition can be easily changed according to the manufacturing conditions of drugs, foods, etc. in which the composition is used.

Example

Hereinafter, the present invention will be described in more detail through examples. These examples are only for exemplifying the present invention, and it will be apparent to those skilled in the art that the scope of the present invention is not to be construed as being limited by these examples.

Example 1: Egg production with increased lutein and zeaxanthin content

58-week-old laying hens were divided into 11 groups, and group 1 was fed 100 g of regular feed daily, and groups 2 to 11 were fed regular feed with Macular pigment 3 (MP3) ( Dunaliella sp. MP3, KCTC 12990BP) algae was supplied at 0.2, 0.4, 0.6, 0.8, 1.2, 1.6, 2.4, 3.2, 4.0 and 5.0% by weight, respectively, and 100 g in total was supplied daily for 15 days. In each experimental group, two laying hens were housed in a space containing a feed trough, water trough, and perch.

The color of egg yolk (Yolk) for eggs produced one week after the start of spawning was compared and shown in FIG. 2. In FIG. 2, the egg yolk of group 5 fed with 0.8% by weight of Dunaliella tertiorecta mutant showed a stronger orange color than the control group fed only with normal feed.

In order to compare the specific chromaticity, 10 g of egg yolk from the control group and the experimental groups fed 0.2, 0.4, and 0.8% by weight were well mixed and spread in a Petri dish, and the chromaticity was measured with a colorimeter, and shown in FIG. 3.

In FIG. 3, there was almost no difference in whiteness and yellowness depending on the experimental group, but as the microalgae were added, the redness gradually increased, confirming that the egg yolk showed a darker color.

In order to measure the lutein and zeaxanthin content of egg yolk, 0.5 g of egg yolk was diluted 20 times with 90% acetone, mixed well, centrifuged, and the supernatant was filtered through a 0.2 μm filter. The results of HPLC analysis are shown in Table 1 below. and shown in FIG. 4 .

Feed rate of microalgae (wt%) Lutein (μg/g) Zeaxanthin (μg/g) 0 10.4 9.9 0.2 12.6 9.6 0.4 15.2 12 0.6 20.5 16.3 0.8 26.9 19.7 1.2 32.1 22.4 1.6 34.9 24.0 2.4 38.9 26.0 3.2 38.4 23.2 4.0 37.6 22.9 5.0 37.3 22.3

In FIG. 4, the content of lutein and zeaxanthin in egg yolk gradually increased as the microalgae mutant was added, but after showing the highest value at 2.4% by weight, it gradually decreased or did not increase anymore. From this, it can be seen that it is not possible to obtain eggs having higher lutein and zeaxanthin contents simply by feeding a large amount of microalgae.

In order to evaluate the efficiency of microalgae feeding, the amount of pigment increase in egg yolk for the fed lutein and zeaxanthin contents (lutein: 6.6 mg/g, zeaxanthin: 3.5 mg/g) was calculated and shown in FIG. 5.

In FIG. 5, the rate of increase of lutein and zeaxanthin increased rapidly as the amount of microalgae was increased, peaked at 0.8% by weight, and then showed a graph in which the efficiency gradually decreased as the amount of addition increased.

Example 2: Egg yolk oil extraction

A test was performed to extract egg yolk oil from the egg yolk prepared in Example 1.

First, 100 mL of ethanol was added to 15 g of egg yolk produced by feeding 2.4 wt% algae, and stirred at 150 rpm for 10 minutes. Thereafter, the mixture was centrifuged, and the supernatant was collected separately, and extraction was repeated until the color of the precipitate became colorless.

The collected supernatant was distilled under reduced pressure to evaporate ethanol and egg yolk oil was obtained. The prepared egg yolk oil is shown in FIG. 6 . Clear yellow lutein/zeaxanthin enriched egg yolk oil was obtained, and the average yield was 23.4%.

To measure the phospholipid content of the prepared egg yolk oil, 5 ml of hexane and 10 ml of cold acetone were added to 1 g of egg yolk oil, centrifuged, and the precipitate was dried and weighed. The phospholipid content of egg yolk oil was 31.5%, which was very high compared to the 1.2% phospholipid content of commercially available egg yolk oil.

The carotenoid content of egg yolk oil was calculated by HPLC analysis after 100 μl of egg yolk oil was diluted 20-fold with THF and filtered through a 0.2 μm filter.

The lutein/zeaxanthin enriched egg yolk oil was measured to contain 0.0979 mg/g and 0.0668 mg/g of lutein and zeaxanthin, respectively, showing very high lutein and zeaxanthin contents.

Example 3: Preparation of egg yolk oil emulsion

Water was added in a weight ratio of 1:4 to the egg yolk oil prepared in Example 2, and stirred at 13,600 rpm for 2 minutes using a homogenizer to prepare an egg yolk oil emulsion.

Commercially available socho method egg yolk oil was added in a water ratio of 1:4 in the same manner and stirred to prepare an egg yolk oil emulsion. The egg yolk oil emulsion of the present invention and the commercially available egg yolk oil emulsion are shown in FIG. 7 in comparison.

It was confirmed that the lutein/zeaxanthin-enriched egg yolk oil emulsion of the present invention extracted with ethanol was pale and clear yellow and had a light egg smell, but the emulsion made from commercially available egg yolk oil was light brown and had a strong heating odor.

In order to test the storage stability of the egg yolk oil emulsion, 20 μl was taken from the bottom immediately after preparing the emulsion, diluted 200 times with distilled water, and the absorbance at 500 nm was measured as A0.

The emulsion was stored at 4, 25, and 37° C. for one day, respectively, and 20 μl was taken from the bottom of the emulsion, diluted 200 times with distilled water, and the absorbance at 500 nm was measured as A1.

In addition, the same amount of solution having the salt concentration as shown in Table 2 below was added to the emulsion, mixed well, stored at 25 ° C. for one day, 20 μl was taken from the bottom of the emulsion and diluted 200 times with distilled water The value measured for absorbance at 500 nm was set as A2.

Emulsion stability (ESI) was calculated as A1/A0 and A2/A0 and is shown in Table 2 below.

ESI Storage temperature (℃) Salt concentration (mM) 4 25 37 50 100 300 Example 0.950 0.900 0.946 0.412 0.289 0.325 comparative example 0.310 0.252 0.175 0.198 0.184 0.153

In the above table, the egg yolk oil emulsion of the present invention has a very high phospholipid content of 31.5%, so the stability against temperature changes is very high, but the commercially available egg yolk oil emulsion has a very low phospholipid content of about 1.2%, so its storage stability according to temperature is very low. It can be seen that it is very unstable.

In addition, in the case of storage stability according to salt concentration, the lutein and zeaxanthin-enriched egg yolk oil emulsion of the present invention showed much better stability than commercially available egg yolk oil emulsion at all salt concentrations.

In order to test the storage stability of the lutein and zeaxanthin contents of the egg yolk oil emulsion, the egg yolk oil emulsion was stored for 5 days under each condition described in Tables 3 to 5 below, and then diluted 10-fold with THF to perform HPLC analysis.

mg/ml 4℃ 25℃ 37℃ lutein 13.7107 13.3724 9.2121 dark condition,
nitrogen substitution zeaxanthin 10.3697 10.0770 6.7297

Table 3 shows the concentration of lutein / zeaxanthin in the emulsion when only the storage temperature is different under the same light and air conditions. At 4 ° C and 25 ° C, there was no significant difference in the contents of lutein and zeaxanthin, but it can be seen that the contents of lutein and zeaxanthin were significantly reduced at 37 ° C.

mg/ml light conditions dark condition lutein 6.0900 13.3733 nitrogen substitution,
25℃ zeaxanthin 4.4733 10.0767

Table 4 shows the concentration of lutein/zeaxanthin in the emulsion when only the light conditions were changed while the air and temperature conditions were the same. It can be seen that lutein/zeaxanthin is more decomposed in the light condition than in the dark condition, and the content is reduced.

mg/ml air exposure nitrogen substitution lutein 10.0000 6.0900 light condition,
25℃ zeaxanthin 7.1067 4.4733

Table 5 shows the concentrations of lutein and zeaxanthin in the emulsion when the light and temperature conditions were the same and only the air conditions were stored differently. It can be seen that lutein and zeaxanthin are less degraded under air exposure conditions.

Example 4: Preparation of egg yolk powder

Egg yolk powder was prepared for commercially available normal eggs and lutein- and zeaxanthin-enriched eggs prepared in Example 1, respectively.

First, the yolk of the egg was separated and each sample was homogenized at 60 ° C. at 25,000 rpm for 5 minutes. After sterilization in a constant temperature water bath at 60 ° C. for 15 minutes, egg yolk powder was prepared by spray drying while maintaining the product temperature at 40 ° C. by adding the excipients shown in Table 6 below.

division Egg yolk (g)
/ DS excipient (g) recovery
(g) recovery rate
(%) G L DP GA DT A 146 / 73 - - - - - 54.91 37.6 eB 220/110 7 7 7 - - 99.64 41.3 eC 220/110 5 4 4 4 4 104.79 43.5

※ In the above table, A: commercially available egg yolk, eB, eC: lutein/zeaxanthin-enriched egg yolk, G: gelatin, L: lactose, DP: starch disphosphate, GA: gum arabic, DT: glusidex (maltodextrin), DS : Purified water

In addition, in order to prepare egg yolk powder by freeze-drying, the egg yolk was sterilized in a constant temperature water bath at 60° C. for 3.5 minutes and then cooled at 0° C. It was put into a sterile receiver and frozen while maintaining at -68 ° C. for 2 hours, and then freeze-dried. The recoveries of lyophilized egg yolk were 51.25% and 46.78% for commercially available egg yolk (F) and lutein/zeaxanthin enriched egg yolk (eF), respectively.

The prepared egg yolk powder is shown in FIG. 8 . 8, it can be seen that the egg yolk powders prepared from lutein/zeaxanthin-enriched egg yolks (eB, eC, and eF) exhibit a darker yellow color than the egg yolk powders prepared from commercially available egg yolks (A, F).

Example 5: Preparation of mayonnaise using egg yolk powder

Mayonnaise was prepared using the five egg yolk powders prepared in Example 4.

After preparing the ingredients in Table 7 below, all other samples except corn oil were first mixed, and then mayonnaise was prepared by homogenizing at 10,000 rpm for 5 minutes while slowly adding corn oil. The prepared mayonnaise is shown in FIG. 9 .

ingredient Percentage (wt%) egg yolk powder 5 corn oil 70 water 15 salt 1.5 sugar 5 vinegar 3.5

9, it can be visually confirmed that mayonnaise prepared using lutein/zeaxanthin-enriched egg yolk powder exhibits a deeper yellow color than mayonnaise prepared using normal egg yolk powder.

In order to test the storage stability of the prepared mayonnaise, the results are shown in FIG. 10 after storing 5 types of mayonnaise for 3 weeks.

In FIG. 10, all five types of mayonnaise maintained emulsion stability for 21 days (3 weeks) or more without oil leakage. Therefore, it was confirmed that mayonnaise prepared using egg yolk powder had excellent emulsion stability and was sufficiently commercialized.

Example 6: Sensory evaluation of mayonnaise

Sensory evaluation of the mayonnaise prepared in Example 5 was performed.

Sensory evaluation was evaluated by 20 panelists aged 20 to 35 years, and mayonnaise samples were provided in a standard amount (5 g) in a disposable cup. All samples were coded with three random numbers and asked to rinse their mouths with bottled water between assessments.

The sensory properties of mayonnaise were rated on a 9-point scale (1 = 'extremely dislike', 9 = 'extremely good'). The results are shown in Table 8 below.

color incense taste stickiness zest overall acceptance A 4.95±1.50 5.05±1.43 4.35±1.93 4.05±1.76 4.10±2.05 4.4±1.70 F 6.00±1.26 4.95±1.54 5.10±1.55 4.40±1.73 5.20±1.58 5.15±1.76 eB 6.45±1.32 5.30±1.38 5.65±1.46 5.85±1.76 5.50±1.61 5.75±1.71 eC 6.45±1.23 6.05±1.47 6.30±1.38 6.10±1.80 6.00±1.62 6.30±1.81 eF 6.40±1.27 6.00±1.62 5.65±1.66 4.80±1.61 5.65±1.79 5.85±1.60

In the table above, mayonnaise prepared with lutein/zeaxanthin-enriched egg yolk powder (eB, eC, eF) showed better sensory evaluation overall than mayonnaise prepared with normal egg yolk powder (A, F).

Among mayonnaises prepared with lutein/zeaxanthin-enriched egg yolk powder, mayonnaise using egg yolk powder (eC) prepared through spray drying using 5 types of excipients showed the best sensory evaluation in all items.

As above, specific parts of the contents of the present invention have been described in detail, and for those skilled in the art, these specific descriptions are only preferred embodiments, and the scope of the present invention is not limited thereby. It will be clear. Accordingly, the substantial scope of the invention will be defined by the appended claims and their equivalents.

Claims (15)

A method for producing eggs with increased content of lutein and zeaxanthin, comprising the step of obtaining eggs by feeding a feed or feed additive containing 1.5 to 2.5% by weight of microalgae containing lutein and zeaxanthin to laying hens,
The microalgae are Dunaliella sp. or genetic mutants thereof,
A method for producing eggs with increased content of lutein and zeaxanthin, wherein the content of lutein in the egg yolk of the produced egg is 34.9 to 38.9 μg/g. delete delete According to claim 1,
Method for producing eggs with increased content of lutein and zeaxanthin, characterized in that the feed or feed additive is fed for 7 to 60 days. delete delete delete delete delete delete delete delete delete delete delete

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