KR20200071028A - Solubilizing composition of insoluble material comprising ligosaccharides prepared using lactic acid bacteria - Google Patents

Abstract

The present invention relates to a solubilizing composition for an insoluble material, the composition comprising oligosaccharides prepared using lactic acid bacteria. More specifically, the composition comprising fermented oligosaccharides according to the present invention significantly increases the levels of solubilization of turmeric powder, pterostilbene, epigallocatechin gallate (EGCG), quercetin, idebenone, and oleanolic acid, which are insoluble materials. Thus, the composition can be advantageously used for solubilizing insoluble materials in aqueous solutions, being applicable to cosmetics, food, and drugs of various kinds.

Description

An aqueous solution solubilizing composition of a poorly water-soluble substance containing an oligosaccharide prepared using a lactic acid bacteria strain {SOLUBILIZING COMPOSITION OF INSOLUBLE MATERIAL COMPRISING LIGOSACCHARIDES PREPARED USING LACTIC ACID BACTERIA}

The present invention relates to an aqueous solution solubilizing composition of a poorly water-soluble substance containing an oligosaccharide prepared using a lactic acid bacteria strain.

Recently, in the cosmetics and food and drug industry, the importance of developing natural ingredients having excellent biocompatibility and biosafety as well as its activity has emerged in order to discover bioactive materials. However, these natural ingredients have different overall characteristics, and thus a formulation capable of sufficiently stabilizing all of them has not been developed.

Currently, natural ingredients that show high efficacy are expected to have great potential, and about 40% of them are not even entering the development stage due to low solubility. The process of improving the solubility by manipulating such a poorly water-soluble substance is called solubilization, and a method for solubilizing the poorly water-soluble substance in water has been developed by many researchers. Since such a solubilization technology has a very large ripple effect on the industry, it is expected that the solubilization technology of a poorly water-soluble material will be further accelerated in the future.

The hydrophobic, poorly water-soluble material is characterized by forming a molecular arrangement that is difficult to solubilize in water due to strong hydrophobic interaction between molecules when the hydrophilic group is absent or very small, or in a position that interferes with the molecular arrangement. If the molecular arrangement of the hydrophobic material can be modified, the poorly water-soluble material may be dispersed or solubilized, and a surfactant capable of inducing a change in the molecular arrangement is a surfactant.

Various surfactants are useful in the cosmetic and food and drug industries. Conventionally, a technique for nanoparticles has been developed in a physical condition using an ultra-high pressure emulsifier (ultrahomogenizer) using synthetic and natural surfactants. However, the above technique has a limitation in solubilizing a poorly water-soluble substance at a high concentration and its stability, and thus, development of a new method is required.

In this regard, Korean Patent Registration No. 10-1138258 discloses a method of forming a structure having a hydrophobic pupil structure using an oligomer derived from two types of hydrophilic natural polymers, and encapsulating a poorly water-soluble substance in the pupil structure to solubilize it. Doing.

Republic of Korea Registered Patent No. 10-1138258

An object of the present invention is to provide an aqueous solution solubilizing composition of a poorly water-soluble substance containing fermented oligosaccharides.

In order to achieve the above object, the present invention provides an aqueous solution solubilizing composition of a poorly water-soluble substance containing fermented oligosaccharides.

In addition, the present invention provides a method for solubilizing an aqueous solution of a poorly water-soluble substance, comprising dissolving a poorly water-soluble substance in the aqueous solution-soluble composition.

Furthermore, the present invention provides a method for producing fermented oligosaccharides comprising inoculating and culturing lactic acid bacteria strains.

The composition comprising the fermented oligosaccharide according to the present invention significantly increases the solubility of aqueous solutions of the water-insoluble substances such as turmeric powder, pterostilbene, epgallocatechin gallate (EGCG), quercetin, idebenone, and oleonolic acid, the composition Can be usefully used for solubilizing aqueous solutions of poorly water-soluble substances applicable to various types of cosmetics and food and drug products.

1 is a photograph of the result of visually confirming the degree of solubilization of an aqueous solution by dissolving turmeric powder in steviol oligosaccharide (Ste-NGOS), fermented oligosaccharide (NGOS), or water, respectively.
2 is a view showing a standard curve obtained by using turmeric powder dissolved in ethanol to quantify the amount of turmeric powder solubilized in aqueous solution.
3 is a graph showing the results of quantifying the amount of turmeric powder solubilized in aqueous solution using a standard curve.
Figure 4 is a result of analyzing the aqueous solution solubilized pterostilbene by TLC method.
5 is a graph showing the result of quantifying the amount of aqueous solution solubilized pterostilbene using the TLC method.
FIG. 6 is a photograph showing results of visually confirming the degree of solubilization of an aqueous solution by dissolving EGCG in fermented oligosaccharide (A) or steviol oligosaccharide (B), respectively.
7 is a view showing the results of analyzing the aqueous solution solubilized EGCG by TLC method.
8 is a photograph showing the result of visually confirming the degree of solubilization of an aqueous solution by dissolving quercetin in fermented oligosaccharide (A) or steviol oligosaccharide (B), respectively.
Figure 9 is a result of dissolving idebenone in steviol oligosaccharide and analyzing the degree of solubilization of aqueous solution by TLC method, and confirming the results under visible light (A) or UV (B).
10 is a photograph showing the result of dissolving idebenone in steviol oligosaccharide and visually confirming the solubility of the aqueous solution.
11 is a view showing the result of analyzing the degree of solubilization of an aqueous solution by dissolving oleonolic acid in steviol oligosaccharide by TLC method.

Hereinafter, the present invention will be described in detail.

The present invention provides an aqueous solution solubilizing composition of a poorly water-soluble substance containing fermented oligosaccharides.

As used herein, the term “fermented oligosaccharide” refers to a water-soluble crystalline substance having a sweetness in which 2 to 8 sugars are bound by synthesis of an enzyme. The fermented oligosaccharide is excellent in physiological functions and physicochemical properties, and can be applied to various food and drug products. In particular, it is mainly used as a substitute for sugar because it has similar physical properties and sweetness to sugar.

The fermented oligosaccharide is glucose, fructose, galactose, sucrose, maltose, maltotriose, lactose, tagatose, allulose, stevioside, stevia, steviol glycoside, mogroside ), any one or more selected from the group consisting of cucurbitane glycoside, phyllodulcin and glycyrrhizin may be fermented. The fermented oligosaccharide may be prepared by a conventional method, or may be prepared by a method appropriately modified by a person skilled in the art as necessary. Specifically, the fermented oligosaccharide may be an oligosaccharide fermented by lactic acid bacteria.

The fermentation may be carried out at a temperature of 25 to 50 ℃, 30 to 50 ℃, 40 to 50 ℃, 25 to 47 ℃, 30 to 47 ℃ or 40 to 47 ℃. In addition, the fermentation for 3 to 20 hours, 3 to 17 hours, 3 to 13 hours, 6 to 20 hours, 6 to 17 hours, 6 to 13 hours, 8 to 20 hours, 8 to 17 hours or 8 to 13 hours Can be performed.

The lactic acid bacteria used for fermentation of the fermented oligosaccharide according to the present invention may include all lactic acid bacteria known in the art. Specifically, the lactic acid bacteria may be strains of the genus Leukonostock , for example, the strains of the genus Leukonostock are Leuconostoc ticreum , Leuconostoc mesenteroide and Leuconostoc mesenteroide Leuconostoc lactis ) may be any one or more selected from the group consisting of.

In one embodiment of the present invention, the leuconostoc mesenteroid strains are leuconostoc mesenteroides B-1355C/KM strain (KCCM 11730P) and leuconostoc mesenteroides B-512F/KM strain ( KCCM 11728P) may be any one or more selected from the group consisting of. The Leukonostock Mesenteroides B-1355C/KM strain and Leukonostock Mesentoroides B-512F/KM strain may be mixed and used in appropriate amounts by a person skilled in the art.

The poorly water-soluble material may include all materials known in the art to be poorly water-soluble materials. For example, the poorly water-soluble material may be any one or more selected from the group consisting of turmeric powder, pterostilbene, epigallocatechin gallate (EGCG), quercetin, idebenone and oleonolic acid.

In addition, the present invention provides a method for solubilizing an aqueous solution of a poorly water-soluble substance, comprising dissolving a poorly water-soluble substance in the aqueous solution-soluble composition.

The aqueous solution solubilizing composition used in the method for solubilizing an aqueous solution of a poorly water-soluble substance according to the present invention may have the characteristics as described above. In one example, the aqueous solution solubilizing composition may include a fermented oligosaccharide. The fermented oligosaccharide may be an oligosaccharide fermented with lactic acid bacteria, and specifically, the lactic acid bacteria may be a strain of genus Leuconostock. The strain of genus Leukonostock may be any one or more selected from the group consisting of Leuconostock citreum, Leuconostock mesenteroid, and Leuconostock lactis.

On the other hand, the water-soluble material capable of increasing water solubility by the aqueous solution solubilization method of the water-soluble material according to the present invention is a group consisting of turmeric powder, pterostilbene, EGCG (epigallocatechin gallate), quercetin, idebenone and oleonolic acid It may include any one or more selected from.

Furthermore, the present invention provides a method for producing fermented oligosaccharides comprising inoculating and culturing lactic acid bacteria strains.

Fermented oligosaccharides prepared by the method for producing fermented oligosaccharides according to the present invention may have the characteristics as described above. In one example, the fermented oligosaccharide is glucose, fructose, galactose, sucrose, maltose, maltotriose, lactose, tagatose, allulose, stevioside, stevia, steviol glycosides, moglosides, cquerbitane glycosides, Any one or more selected from the group consisting of pilodulcine and glycyrrhizin may be fermented. Specifically, in one embodiment of the present invention, the oligosaccharide can be prepared by mixing sucrose or stevioside with maltose and sodium acetate buffer, and inoculating it with lactic acid bacteria.

The lactic acid bacteria strain may be a strain of genus Leukonostock. The strain of genus Leukonostock may be any one or more selected from the group consisting of Leuconostock citreum, Leuconostock mesenteroid, and Leuconostock lactis.

The fermentation can be performed under conventional conditions, and can be carried out by appropriate modification by a person skilled in the art. Specifically, the fermentation can be carried out at a temperature of 25 to 50 ℃, 30 to 50 ℃, 40 to 50 ℃, 25 to 47 ℃, 30 to 47 ℃ or 40 to 47 ℃. In addition, the fermentation for 3 to 20 hours, 3 to 17 hours, 3 to 13 hours, 6 to 20 hours, 6 to 17 hours, 6 to 13 hours, 8 to 20 hours, 8 to 17 hours or 8 to 13 hours Can be performed.

Hereinafter, the present invention will be described in detail by the following examples, provided that the following examples are only for illustrating the present invention, and the present invention is not limited by them. Anything that has substantially the same configuration and the same operation and effect as the technical idea described in the claims of the present invention is included in the technical scope of the present invention.

Example 1. Preparation of fermented oligosaccharide

Fermented oligosaccharides were prepared as follows.

First, leuconostoc mesenteroides ( leuconostoc mesenteroides ) B-1355C/KM strain (KCCM 11730P) (hereinafter,'B-1355C strain') and leuconostoc mesenteroides B-512F/KM strain (KCCM 11728P) ( Hereinafter,'B-512F strain') was cultured by a conventional method to prepare each culture solution. 1 unit/ml B-1335C strain culture, 7 unit/ml B-512F strain culture, 2 M sucrose, 0.25 M maltose and 20 mM sodium acetate buffer (pH 5.2) at 45° C. By reacting for 10 hours, fermented oligosaccharide (NGOS) was prepared.

Example 2. Preparation of steviol oligosaccharide

Instead of sucrose, steviol oligosaccharide (Ste-NGOS) was prepared in the same conditions and method as in Example 1, except that 3% (w/v) of stevioside (Daepyeong, Korea) was added.

Experimental Example 1. Solubilization of aqueous solution of turmeric powder

1-1. Preparation of solution containing aqueous solution solubilized turmeric powder

Using the prepared fermented oligosaccharides and steviol oligosaccharides, turmeric powder was solubilized in an aqueous solution as follows.

Specifically, fermented oligosaccharide or steviol oligosaccharide was added to 10% (w/v) turmeric powder. At this time, water was added to the control group. To this, ethanol was added to a final concentration of 30% (v/v), which was stirred for 1 hour. The stirred material was centrifuged for 30 minutes under conditions of 12,000 rpm and 4° C. to obtain a supernatant, and ethanol present in the obtained supernatant was evaporated to finally obtain a solution containing turmeric powder solubilized in aqueous solution. The results obtained by visually checking the obtained solution are shown in FIG. 1.

As shown in FIG. 1, the water solubility of turmeric powder was high in a solution prepared using fermented oligosaccharides and steviol oligosaccharides. On the other hand, turmeric powder hardly melted in the solution prepared using the control water.

1-2. Quantitative confirmation of turmeric powder water solubility

By performing a thin layer chromatography (TLC) using the solution containing the aqueous solution solubilized turmeric powder prepared in Experimental Example 1-1, the amount of the aqueous solution solubilized turmeric powder was quantitatively confirmed. At this time, when the aqueous solution was solubilized by adding fermented oligosaccharide or steviol oligosaccharide as a control, a solution without adding 30% (v/v) ethanol was prepared and used.

TLC was performed by a conventional method using a fixed phase silica gel 60, a mixed solvent in which acetonitrile and water were mixed in a volume ratio of 85:15 as a developing solvent. The absorbance of each fraction obtained through TLC was measured, and the result obtained by quantitatively obtaining the amount of turmeric powder solubilized in aqueous solution was applied to the standard curve (FIG. 2) obtained with the turmeric powder solution dissolved in ethanol. Shown.

As shown in Fig. 3, when solubilizing turmeric powder in aqueous solution, the amount of turmeric powder solubilized in aqueous solution was significantly increased compared to the case where 30% (v/v) of ethanol was added.

Experimental Example 2. Confirmation of the degree of solubilization of aqueous solution of pterostilbene

2-1. Preparation of solution containing aqueous solution solubilized pterostilbene

Pterostilbene was added instead of turmeric powder, and a solution containing pterostilbene solubilized in aqueous solution was obtained in the same manner and conditions as in Preparation Example 1, except that water and DMSO were used as controls.

2-2. Quantitative confirmation of pterostilbene water solubility

TLC was performed in the same method and conditions as described in Example 1-2 using the solution containing aqueous solution solubilized pterostilbene prepared in Experimental Example 2-1, and the TLC analysis results are shown in FIG. 4, AlphaEaseFC 4.0. The results of quantitatively obtaining the amount of aqueous solution solubilized pterostilbene using the program are shown in FIG. 5. At this time, the amount of pterostilben solubilized in aqueous solution was calculated based on the value of the sample in which sterostilbene was dissolved in DMSO.

As shown in Figure 4, pterostilbene was dissolved in a solution solubilized in aqueous solution using fermented oligosaccharides and steviol oligosaccharides.

5, it was confirmed that 2.21 mg/ml of pterostilben was dissolved in a solution solubilized in an aqueous solution using steviol oligosaccharide.

Experimental Example 3. Solubilization of aqueous solution of EGCG (epigallocatechin gallate)

3-1. Preparation of solution containing aqueous solution solubilized EGCG

Preparation Example 1, except that 100 mg/ml of EGCG was added instead of turmeric powder and fermented oligosaccharide or steviol oligosaccharide was added to be 0, 10, 30, 50, 70 or 100% (v/v). A solution containing EGCG solubilized in an aqueous solution was obtained under the same method and conditions. The result of solubilization of the aqueous solution of EGCG in the prepared solution was visually confirmed and the results of the photographing are shown in FIG. 6.

As shown in Fig. 6, the solubilization of the aqueous solution of EGCG increased depending on the concentration of the fermented oligosaccharide or steviol oligosaccharide used.

3-2. Increased EGCG water solubility

TLC was performed in the same method and conditions as described in Example 1-2 using the solution containing the aqueous solution solubilized EGCG prepared in Experimental Example 3-1, and the results are shown in FIG. 7. At this time, as a control, a solution of 10, 20, 30, 50, 70 or 100 mg/ml EGCG dissolved in ethanol was used.

As shown in Figure 7, the solubilization of the aqueous solution of EGCG increased depending on the concentration of fermented oligosaccharides used.

Experimental Example 4. Solubilization of aqueous solution of quercetin

4-1. Preparation of solution containing aqueous solution solubilized quercetin

Preparation Example 1, except that 500 mg/ml of quercetin was added instead of turmeric powder and fermented oligosaccharide or steviol oligosaccharide was added to be 0, 10, 30, 50, 70 or 100% (v/v) A solution containing quercetin solubilized in an aqueous solution was obtained under the same method and conditions. The result of solubilization of the aqueous solution of quercetin in the prepared solution was visually confirmed and the results of the photographing are shown in FIG. 7.

As shown in FIG. 8, the aqueous solution solubilization of quercetin increased depending on the concentration of the fermented oligosaccharide or steviol oligosaccharide used, and particularly the aqueous solution solubilization of quercetin in steviol oligosaccharide was further increased.

4-2. Quantitative confirmation of quercetin water solubility

TLC was performed in the same method and conditions as described in Example 1-2 using the solution containing the aqueous solution solubilized quercetin prepared in Experimental Example 4-1. On the other hand, the amount of quercetin solubilized in aqueous solution is shown in Table 1 below using a standard curve obtained by using a solution in which quercetin is dissolved in DMSO.

0% (v/v) 10% (v/v) 30% (v/v) 50% (v/v) 70% (v/v) 100% (v/v) Degree of solubilization to fermented oligosaccharide (mg/ml) ND ND 0.02 0.03 0.05 0.14 Degree of solubilization to steviol oligosaccharide
(Mg/ml) ND 0.02 0.07 0.25 0.41 3.12

As shown in Table 1, as the concentration of fermented oligosaccharides or steviol oligosaccharides increased, solubilization degree of quercetin also increased.

Experimental Example 5. Solubilization of aqueous solution of idebenone

5-1. Confirmation of increased water solubility of idebenone

1% (w/v) of idebenone was added to 0, 10, 30, 50 or 70% (v/v) of steviol oligosaccharide and stirred sufficiently for 1 hour to obtain a mixture. The mixture was centrifuged at 12,000 rpm and 4° C. for 20 minutes to take only the supernatant. TLC was performed using the supernatant in the same method and conditions as described in Example 1-2, and the results are shown in FIG. 9.

As shown in Fig. 9, solubilization of the aqueous solution increased depending on the concentration of steviol oligosaccharide in which idebenone was used.

5-2. Quantitative confirmation of idebenone water solubility

From the TLC results performed in Experimental Example 5-1, the amount of idebenone accommodated in steviol oligosaccharide was quantitatively confirmed using the AlphaEaseFC 4.0 program (Alpha Inotech, USA). At this time, the amount of idebenone solubilized in aqueous solution is shown in the following Table 2 with a standard curve obtained using a solution of idebenone dissolved in DMSO. In addition, the solubilization of the aqueous solution of idebenone in the prepared solution was visually confirmed and the results of the photographing are shown in FIG. 10.

0% (v/v) 10% (v/v) 30% (v/v) 50% (v/v) 70% (v/v) Degree of solubilization to steviol oligosaccharide (mg/ml) ND ND ND ND 10.3

As shown in Table 2 and FIG. 10, idebenone was dissolved at a concentration of 10.3 mg/ml in 70% (v/v) steviol oligosaccharide.

Experimental Example 6. Solubilization of aqueous solution of oleanolic acid

The experiment was conducted under the same conditions and methods as in Experimental Example 5, except that oleonolic acid was used instead of idebenone, and solubilization of the aqueous solution of oleonolic acid was confirmed. The results of TLC analysis are shown in FIG. 10, and the result of solubilization of the aqueous solution of oleonolic acid is visually shown in FIG. 11, and the amount of oleonolic acid solubilized in the aqueous solution is shown in Table 3 below.

0% (v/v) 10% (v/v) 30% (v/v) 50% (v/v) 70% (v/v) Degree of solubilization to steviol oligosaccharide (mg/ml) ND 1.0 1.6 2.8 2.3

As shown in FIG. 10, as a result of TLC, solubilization of the aqueous solution increased depending on the concentration of steviol oligosaccharide in which oleonolic acid was used. This, as shown in Table 3 and Figure 11, was confirmed more specifically in the result of quantifying the amount of aqueous solution solubilized oleonolic acid, shows the highest degree of solubilization in 50% (v / v) of steviol oligosaccharides , In the steviol oligosaccharide having a concentration higher than that, it was confirmed that the degree of solubilization was saturated.

Claims (9)

An aqueous solution solubilizing composition of a poorly water-soluble substance containing fermented oligosaccharides.
The method of claim 1, wherein the fermented oligosaccharide is glucose, fructose, galactose, sucrose, maltose, maltotriose, lactose, tagatose, allulose, stevioside, stevia, steviol glycosides , Any one or more selected from the group consisting of mogroside, cucurbitane glycoside, phyllodulcin, and glycyrrhizin, fermented aqueous solubility aqueous solution composition.
According to claim 1, wherein the fermentation is fermentation by lactic acid bacteria, aqueous solubilizing composition of a poorly water-soluble substance.
According to claim 3, wherein the lactic acid bacteria are strains of the genus Leuconostock, aqueous solubilizing composition of a poorly water-soluble substance.
The method of claim 4, wherein the strain of the genus Leuconostoc is any one selected from the group consisting of Leuconostoc ticreum , Leuconostoc mesenteroide , and Leuconostoc lactis . The aqueous solution solubilizing composition of the above-mentioned water-insoluble substance.
According to claim 1, wherein the water-insoluble material is turmeric powder, pterostilbene, EGCG (epigallocatechin gallate), quercetin, idebenone, and any one or more selected from the group consisting of oleonolic acid, aqueous solubilizing composition of the water-soluble material .
A method for solubilizing an aqueous solution of a poorly water-soluble substance, comprising dissolving the poorly water-soluble substance in the aqueous solution-solubilizing composition of claim 1.
Method of producing a fermented oligosaccharide comprising the step of inoculating and culturing the lactic acid bacteria strain.
The method of claim 7, wherein the lactic acid bacteria strain is a strain of genus Leuconostock.

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