KR20000036315A - Process For Preparing Oligosaccharide Containing Sialic Acid - Google Patents

Abstract

PURPOSE: Oligosaccharide containing sialic acid is prepared economically by being extracted and purified from egg yolk without demineralization using ultra filter membrane or ion exchange resin or treatment of protease. CONSTITUTION: Yolk powder is defatted by using an alcohol and the defatted yolk is dried until the content of alcohol is about 5-50 wt.%, then 3-10 times, desirably 6-7 times, of water based on the weight of the defatted yolk is added and mixed. The pH of the solution is adjusted to pH 4-6 to increase a yield by acid hydrolysis as adding hydrochloride for foods then the oligosaccharide containing sialic acid from the solution is extracted.

Description

Process For Preparing Oligosaccharide Containing Sialic Acid

The present invention relates to a manufacturing method for extracting and purifying sialic acid-containing oligosaccharides contained in egg yolk, and more particularly, using ethanol, water, and acid in degreasing egg yolk obtained after extraction and removal of egg yolk oil using a solvent from egg yolk. By providing a method for economically producing a product containing 10 to 20% by weight of sialic acid-containing oligosaccharides.

According to a paper published in J. Carbohydrate Chemistry, 14 (6), 833-841 (1995) by Mamoru Kogetsu et al., Sialic acid in egg yolk is combined with oligosaccharides, and this oligosaccharide is combined with protein sialyl glyco It exists in the form of peptide (Sialylglycopeptides). It is therefore difficult to extract simply with water and can be extracted with proteolytic enzymes or strong ionic liquids.

Japanese Patent Laid-Open No. 6-245784 discloses a method for producing yolk-enzyme-treated sialic acid and sialic acid derivative compositions obtained by enzymatically treating defatted egg yolk, removing the polymer component by ultrafiltration with water-soluble ash, and then performing desalting treatment. Doing.

However, this method has a disadvantage in that the manufacturing cost is expensive because the protein is liquefied by adding a large amount of protease to degreasing egg yolk, and the purity of sialic acid is relatively low. In addition, it causes a secondary problem that it is difficult to industrially use insoluble proteins remaining after preparation.

Japanese Patent Application Laid-Open No. Hei 8-99988 discloses a method for producing sialic acid-containing oligosaccharides by extracting them with water or a salt solution in defatted egg yolk, but desalting a solution (extract) obtained by treating the defatted egg yolk with water or a salt solution. An additional process is needed. The desalting process takes a very long time, such as reverse osmosis, and has the disadvantage of enzymatically treating the extract separately to increase the purity or yield. In particular, the enzyme treatment process requires a long time reaction, and there is a possibility of contamination by microorganisms from the inside and the outside, which requires careful attention. In addition, since the enzyme activity temperature must be maintained at a constant temperature, it is economically expensive and there is a problem in commercial mass production.

Accordingly, the present invention is to provide a method for economically producing a high concentration of sialic acid-containing oligosaccharides industrially economical without the desalting process and protease treatment process using the ultrafiltration membrane or ion exchange resin of the prior art. As a result, the egg yolk powder is degreased using an alcohol solvent to dry the remaining skim yolk to contain about 5 to 50% by weight of alcohol, mixed with water, and the yield is increased by acid decomposition of the protein. By adjusting the pH to 4 to 6 using an acid to raise, the sialic acid-containing oligosaccharides can be industrially used without desalination and enzymatic treatment.

The present invention is to degrease using an alcohol solvent in egg yolk powder and to dry the remaining skim egg yolk to contain about 5 to 50% by weight of alcohol, to add water to the mixture and to increase the yield through acid decomposition of the protein By adjusting the pH to 4 to 6 with a harmful acid to extract an industrially available sialic acid-containing oligosaccharide without desalination and enzymatic treatment.

The content of ethanol in the defatted egg yolk is preferably contained in an amount of 5 to 50% by weight, and most preferably in an amount of 15 to 30% by weight.

When water is added to the defatted egg yolk containing 5 to 50% by weight of ethanol, the amount of water is preferably added to 3 to 10 times 300 liters to 1,000 liters based on 100 kg of the defatted egg yolk, and most preferably. Preferably, 6 to 7 times of water is added.

On the other hand, in the present invention to adjust the pH with hydrochloric acid for foods used to increase the yield through acid decomposition of the protein, the pH is preferably adjusted to 3-6.

Hereinafter, the present invention will be described in more detail with reference to Examples and Comparative Examples. However, these examples are only to aid the understanding of the present invention, and in any sense does not limit or limit the scope of the present invention.

Example 1

1,200 liters of water is added to 300 kg of defatted egg yolk containing 20 wt% ethanol, and the pH is adjusted to 4 with hydrochloric acid for food additives and stirred for 2 hours. After stirring, the mixture was filtered with a centrifugal dehydrator, the filtrate was concentrated under reduced pressure, and freeze-dried to obtain 28.8 kg of sialic acid-containing oligosaccharide.

The sialic acid content was analyzed to be 17.5% (w / w) by the resorcinol method.

In order to compare the effects of ethanol, the result of the same method using degreasing egg yolk containing 0 to 50% by weight of ethanol is shown in [Table 1].

Example 2

2,100 liters of water is added to 300 kg of defatted egg yolk containing 30% ethanol, and the pH is adjusted to 5 with hydrochloric acid for food additives and stirred for 3 hours. After stirring, the mixture was filtered with a centrifugal dehydrator, and the filtrate was concentrated under reduced pressure and freeze-dried to obtain 30.2 kg of sialic acid-containing oligosaccharide.

The sialic acid content was determined to be 19.3% (w / w) as measured by Resorcinol.

In order to compare the effect of the pH adjusted to pH 3 to 7 prepared by the same method is [Table 2].

[Table 1] Distribution of Sialic Acid-Containing Oligosaccharides According to Ethanol Content

Ethanol Content (%) 0 10 20 30 40 50 Solid content in defatted egg yolk (㎏) 240 240 240 240 240 240 Sialic acid content (%) 13.6 16.1 17.5 17.0 15.8 14.0 Quantity (kg) 25.0 26.1 28.8 31.7 32.6 32.6 Yield (%) 10.4 10.9 12.0 13.2 13.6 13.6

[Table 2] Contents and yields of sialic acid-containing oligosaccharides according to pH change

pH 3 4 5 6 7 Solid content in defatted egg yolk (㎏) 210 210 210 210 210 Sialic acid content (%) 16.2 17.0 19.3 16.8 16.5 Quantity (kg) 25.2 27.7 30.2 29.0 28.4 Yield (%) 12.0 13.2 14.4 13.8 13.5

[Table 3] Comparison of well-known citation technology (comparative example) and the present invention (example)

division Japanese Laid Open Patent Japanese Patent Application Publication 8-99988 Example 1 Example 2 Comparative Example 1 Comparative Example 2 Comparative Example 3 Comparative Example 4 Comparative Example 5 Comparative Example 6 Comparative Example 7 Solid content in defatted egg yolk (㎏) 1,000 1,000 0.01 100 100 100 One 240 210 Protease (kg) 5.0 8.0 - - - 0.03 - - - Ultrafiltration membrane use use none use use use use none none Desalination practice practice practice practice practice practice practice Never Never Sialic acid content (%) 7.5 12 13.1 15 19 11 19 17.5 19.3 Quantity (kg) 300 200 0.0011 9.5 3.9 15.0 0.05 28.8 30.2 yield(%) 30 20 11 9.5 3.9 15 5 12.0 14.4

As can be seen from [Table 3], the method of Japanese Patent Laid-Open No. 6-245784 adds a step of decomposing a large amount of protease to defatted egg yolk and decomposing it with an ultrafiltration membrane, but the purity of sialic acid-containing oligosaccharides is increased. Compared to the present invention, there is a disadvantage in that a long reaction time and an expensive enzyme and ultrafiltration system are required.

In addition, the manufacturing method of Japanese Patent Application Laid-open No. Hei 8-99988 is extracted with water or salt solution in defatted egg yolk, or purified by ultrafiltration membrane using protease in extract, but the purity and yield of sialic acid-containing oligosaccharides is similar to the present invention. The result was obtained.

The sialic acid-containing oligosaccharide prepared by the present invention is a method capable of producing industrially economical high concentrations of sialic acid-containing oligosaccharides without desalting or proteolytic enzyme treatment using ultrafiltration membranes or ion exchange resins used in the prior art. Can be provided.

Claims (6)

Method of preparing sialic acid-containing oligosaccharides by extracting with alcohol and degreasing yolk containing alcohols with water and acid, followed by freeze drying or spray drying. The method for producing sialic acid-containing oligosaccharide according to claim 1, wherein the degreasing egg yolk contains 5 to 50% by weight, most preferably 15 to 30% by weight of alcohols. The method for producing a sialic acid-containing oligosaccharide according to claim 1, wherein the alcohol is ethanol. The method for preparing sialic acid-containing oligosaccharides according to claim 1, wherein the amount of water to be added to the defatted egg yolk is added three to ten times as much as 100 kg of the defatted egg yolk. The method for preparing sialic acid-containing oligosaccharide according to claim 1, wherein the pH is adjusted to 3 to 6 with hydrochloric acid for food used to increase the yield through acid decomposition of the protein. Sialic acid-containing oligosaccharide prepared by the method of claim 1.