KR100434674B1 - The High-Degree Refining Method Of β-Glucan - Google Patents

Abstract

The present invention relates to a high purification method of beta glucan, in the purification of beta glucan by supercritical fluid purification using carbon dioxide and cosolvent, the low purity beta glucan-containing material is added to a solution of ethanol and water After pretreatment by stirring for 30 minutes-5 hours at a temperature of 30-60 ℃, it is characterized in that the purification in a supercritical state of the temperature of 40-80 ℃, pressure conditions of 5200-6600psi, the purification method of the present invention Through this, high purity beta glucan with a content of more than 85% of beta glucan can be obtained, shorten the operation time of the supercritical refining step, which requires a lot of operation, and harden the powder frequently occurring in the supercritical refining step. It can prevent.

Description

The High-Degree Refining Method Of β-Glucan

The present invention relates to a high purification method of beta glucan, and more particularly, to a method for purifying 1,3 / 1,6 beta glucan with high purity using a supercritical fluid extraction method.

1,3 / 1,6 beta glucan was first discovered in Japan in 1964 and is known to exert a wide variety of physiological activities such as immune enhancing, antioxidant, anticancer and skin regeneration.

Such 1,3 / 1,6 beta glucan is a polymer most present in the cell wall of the yeast and serves to support the structure of the yeast by strengthening the structure of the yeast cell wall. The cell wall consisting of beta glucans is insoluble and contains cellular internals.

Representative of the conventional methods of purifying beta glucan from yeast cell walls is a method utilizing differences in solubility in acid and alkaline solutions. In the case of using an acidic solution, acetic acid solution, hydrochloric acid or phosphoric acid is used as the chemical component, and sodium hydroxide is mainly used in the process using an alkaline solution.

The method using such an acid or alkaline solution has a disadvantage in that it is difficult to selectively separate only beta glucan from the cell wall of the yeast cell complex in which beta glucan is complex with each other. For this reason, in order to purify feed yeast having a purity of about 30% of beta glucan to an edible product having a purity of about 60%, the purification method should be repeated several times. In addition, it is impossible to obtain products having a purity of 67% or more with this method.

The physiological activity of beta glucan is related to the molecular weight and purity of beta glucan, and the adverse effects on the human body are deeply related to the purity of beta glucan. Therefore, although it is very important to increase the purity of beta glucan, technical research on this is still insignificant.

As a technique which overcomes the prior art to some extent as described above, Korean Patent Application No. 2001-22650 by the present applicant is given. It describes a method of purifying beta glucan using supercritical fluid purification using carbon dioxide and a cosolvent (alcohol, etc.) as a solvent and at a temperature of 40-80 ° C. and a pressure of 3000-5000 psi.

Supercritical Fluid Purification means that if a substance is maintained at a pressure and temperature above its critical point, it is present as a 'supercritical fluid' substance that is physically and chemically distinct from solids, liquids, and gases. Supercritical fluid purification is a method of purifying a desired substance by using.

This supercritical fluid has a density similar to that of liquid and exhibits properties close to gas in terms of diffusivity and permeability. This supercritical fluid purification technology has been applied to medicine, processed food, and petrochemical fields since the 1970s.

The prior art using a supercritical fluid not only increases the purification efficiency, but also has an environmentally friendly advantage because there is no discharge of waste liquid. However, the technology is somewhat insufficient in purification efficiency, the purification time is relatively long, as well as the beta glucan powder is hardened after a certain time during purification has a problem that the workability is greatly reduced.

The present invention is to solve the problems of the prior art as described above, and an object of the present invention is to provide a method for purifying beta glucan having high purification efficiency and reduced purification time. Another object of the present invention is to prevent the beta glucan powder from hardening during operation.

In the present invention, in purifying beta glucan by supercritical fluid purification using carbon dioxide and cosolvent, the low purity beta glucan-containing substance is added to a solution of ethanol and 1-40% by volume of water at 30-60 ° C. After pretreatment by stirring for 30 minutes-5 hours at a temperature of, characterized in that the purification in a supercritical state of the temperature of 40-80 ℃, pressure conditions of 5200-6600psi.

Hereinafter, the present invention will be described in detail.

In the pretreatment process of the purification method of the present invention, the low-purity beta glucan-containing material is added to a solution of ethanol and 1-40% by volume of water and stirred for 1-5 hours at a temperature of 30-60 ° C., thereby As a result, the water-soluble substance and the fat-soluble substance corresponding to the impurities in the low purity beta glucan-containing substance are removed to some extent, and the specific surface area of the powder of the beta glucan-containing substance is increased.

Powders with an increased specific surface area have high purification efficiency, and materials removed in the pretreatment step are relatively easy to remove. If you go directly to the supercritical purification step without going through the pretreatment step, these easy-to-remove materials not only reduce the purification efficiency of the supercritical step, but do not matter on laboratory scales with chamber volumes within 100 ml. In production, when a certain time (about 2-3 hours) passes in the supercritical purification step, the powder of the beta glucan-containing substance hardens. When the powder is hardened, the purification efficiency is greatly reduced.

In the pretreatment step, the stirring may be performed by a mechanical method, or may be stirred using ultrasonic waves. In terms of efficiency, agitation by ultrasonic waves is somewhat superior. Pretreatment time is appropriate for 1-5 hours, even if this range is exceeded, there is no increased effect.

After the pretreatment is completed, the supercritical purification step is entered, and suitable conditions for the supercritical purification step of the present invention are a temperature of 40-80 ° C. and a pressure condition of 5200-6600 psi. The pressure condition in the prior art was 3000-5000 psi, but when the pretreatment step was performed, the pressure condition of 5400-6000 psi was more preferable than 5200-6600 psi. If the pressure is lower than the above range, the purification efficiency is lowered. If the pressure is higher than the above range, no further increase of the purification efficiency occurs, but only the production cost increases.

Carbon dioxide is used as the solvent, alcohol and the like is used as the co-solvent is the same as the prior art. Carbon dioxide is recycled after being transferred to the separation tank through a pressure reducing valve.

An embodiment of the present invention is as follows.

* Purification method of beta glucan

Purity of the method of purity of beta glucan obtained in the following example was added 100 mg of 1,3 / 1,6 beta glucan to 80 ml of trifluoroacetic acid and then subjected to ultrasonic stirring for 10 minutes and reacted under normal pressure in an autoclave for 1 hour to completely hydrolysis. After confirming that the hydrolyzed solution was taken and quantified by measuring the content of glucose in HPLC.

<Example 1>

A low purity beta glucan powder having a content of about 66.3 (± 3.0)% of beta glucan was added to an ethanol solution containing 20% by volume of water, pretreated for 1 hour while maintaining a temperature of 40 ° C, and carbon dioxide was used as a solvent. 20 vol% ethanol is used as a cosolvent, and it is placed in a supercritical purification chamber (chamber volume 4ℓ) at a temperature of 50 ° C. and a pressure of 5600 psi and purified for 3 hours. It was like Powder hardening did not occur during purification.

Comparative Example 1

The same procedure as in Example 1 was performed except that the pretreatment step was omitted. Beta glucan content of the obtained material was as shown in Table 1 below, and the hardening of the powder began to occur 2 hours after the supercritical purification step was started.

<Example 2>

In the same manner as in Example 1, the solution used in the pretreatment step was changed to an ethanol solution containing 40% by volume of water. Beta glucan content of the obtained material was shown in Table 1 below.

<Example 3>

In the same manner as in Example 1, but the pressure of the supercritical purification chamber was changed to 6500 psi, the beta glucan content of the obtained material was shown in Table 1 below.

Comparative Example 2

In the same manner as in Example 1, the solution used in the pretreatment step was changed to an ethanol solution containing 50% by volume of water. Beta glucan content of the obtained material was shown in Table 1 below.

Comparative Example 3

In the same manner as in Example 1, but the pressure of the supercritical purification chamber was changed to 7000 psi, the beta glucan content of the obtained material was shown in Table 1 below.

division Beta Glucan Content (%) Stiffness Example 1 86.4 (± 3.0) none Example 2 85.8 (± 3.0) none Example 3 86.9 (± 3.0) none Comparative Example 1 75.5 (± 3.0) Occur Comparative Example 2 79.8 (± 3.0) none Comparative Example 3 86.7 (± 3.0) none

As can be seen from the above examples and comparative examples, through the purification method of the present invention it is possible to obtain a high purity beta glucan content of more than 85% of the beta glucan, operation of a supercritical purification step that requires a lot of operation The time can be shortened, and the hardening of the powder frequently occurring in the supercritical purification step can be prevented.

Claims (4)

In purifying beta glucan by supercritical fluid purification using carbon dioxide and cosolvent, low-purity beta glucan-containing substance was added to a mixture of ethanol and water for 30 minutes-5 hours at a temperature of 30-60 ° C. After pretreatment by stirring, the purification method of beta glucan, characterized in that the purification in a supercritical state at a temperature of 40-80 ℃, pressure conditions of 5200-6600psi. The method of claim 1, wherein the cosolvent is beta glucan advanced purification method, characterized in that at least one selected from ethanol, methanol, water. The method of claim 1, wherein the supercritical pressure condition is 5400-6000 psi. The method of claim 1, wherein the mixed solution of ethanol and water is 1-40% by volume of water in ethanol.