KR20170119571A - A method for extracting low molecular weight β-glucan from barley - Google Patents

Abstract

The present invention relates to a method for extracting low molecular weight? -Glucan from barley, and more particularly, to a method for extracting low molecular? -Glucan from barley capable of effectively extracting and extracting low molecular weight? -Glucan from a barley cell wall, a) pulverizing barley to obtain a barley powder; b) adding hot water to the barley powder and stirring to swell the? -glucan; c) treating β-glucan swelled barley powder suspension with protease and? -amylase to extract? -glucan.

Description

The present invention relates to a method for extracting low-molecular β-glucan from barley,

The present invention relates to a method for extracting low molecular β-glucan from barley, and more particularly, to a method for extracting low molecular β-glucan from barley capable of effectively extracting and extracting low molecular weight β-glucan from barley cell walls.

β-Glucan is a water-soluble polysaccharide that is linked to β-1,2-1,4-linked straight chains and contains water-soluble polysaccharides of the same form in oats. However, glucan is the most abundant in cereals It is barley.

Β-Glucan contained in barley is evaluated as an effective ingredient for the prevention of lifestyle-related diseases and has been proved effective in Korea, Japan, USA, Canada, Europe, Australia and New Zealand, and its function is also permitted .

Even if water-soluble? -Glucan is ingested, the human body does not digest an enzyme capable of digesting dietary fiber and reaches the small intestine. And then digested by intestinal bacteria to produce various physiologically active substances such as short chain fatty acids. There are hundreds to thousands of germs in the intestinal tract and there are about 100 trillions of germs in the intestinal flora. These intestinal bacterial populations include good bacteria such as Bifidus, lactobacillus, and yeast that are effective in maintaining human health and harmful bad bacteria such as Clostridium and pathogenic Escherichia coli , Bacteroides bacteria, and non-pathogenic Escherichia coli, which are known as opportunistic pathogens.

Recently, the pace at which research has progressed in relation to the study of bacterial populations in the intestinal tract and the health of the human body has surprisingly progressed to include immune system, metabolic syndrome, diabetes mellitus, allergies, And immune system diseases such as major dispression are also pointed out to be caused by the imbalance of intestinal bacterial populations and it is pointed out that it is important to maintain the balance of intestinal bacterial populations.

The main ingredient of barley is about 80% starch. The insoluble portion remaining after the separation of? -glucan contains approximately 80% of the fraction and 4% of insoluble dietary fiber. The insoluble dietary fiber and oligosaccharide obtained from the barley by heating the insoluble fraction and heating the starch to α-amylase and then transferring it to the oligosaccharide using β-amylase and Trans-glucosidase, It becomes a useful material to maintain or improve the balance of the group.

On the other hand, various methods have been proposed in connection with a method of extracting? -Glucan from cereals such as barley.

Patent Literature 0001 relates to a method for extracting? -Glucan from cereals, comprising: a first step of finely treating a cereal containing? -Glucan and adding water to the powder to extract an aqueous solution; A second step of heating the extracted aqueous solution to inactivate? -Glucanase, coagulating the protein, adding a starch hydrolyzing enzyme to the reaction solution, and separating a solution except for the solidified substance by filtration; And a third step of separating the solid? -Glucan by adding alcohol or alcohol to the separated and separated solution.

Patent Literature 0002 relates to a method for producing a high-purity and high-purity beta-glucan from barley, wherein an increase in extraction yield and inactivation of beta-glucanase are simultaneously achieved by treating the barley powder with hydrogen peroxide and heating, In order to prevent further decomposition of beta-glucan by hydrogen peroxide, catalase and phytic acid are added as hydrogen peroxide inhibitors, and alpha-amylase treatment is performed to remove residual polysaccharides and oligosaccharides to prepare water-soluble beta-glucan in high purity and high yield .

Patent Document 0004 relates to a barley culture medium and a method for producing beta-glucan from Shoshophylum corn using the barley culture medium, which comprises culturing a Shyozypiumum corneum strain in a barley medium containing 0.5 to 1.5% by weight of barley and 0 to 1.0% To produce? -Glucan.

Patent Document 0004 relates to a method for extracting β-glucan, which is characterized by obtaining barley or oat-derived water-soluble β-glucan by hot water extraction of barley rice bran, oat rice bran, crushed non-veined barley grain or crushed marine oat grain.

Patent Document 0005 relates to a lipid metabolic pathway gift and a method for producing the same, wherein an extract obtained by extracting oats or barley with an alkaline aqueous solution is precipitated by acid precipitation of the protein, and then the alcohol is added to the remaining solution to precipitate or the residue is desalted and dried Glucan as a main component.

Patent Document [0006] The present invention relates to a low-molecular-weight? -Glucan having an immunopotentiating action and, as a drug substance for immunomodulating action, a water-soluble? -Glucan derived from a barley powder whose weight average molecular weight is 5,000 to 100,000, .

Patent Document 0007 is directed to a method for producing β-glucan and β-glucan, and is characterized by providing a method for producing low-molecular-weight β-glucan derived from grains having a step of hydrolyzing enzyme.

(A) Water-soluble β-1,3-1,4-glucan having a molecular weight of 500,000 or more (B) Water-soluble β-1,3-glucan having a molecular weight of 20,000 or more and 500,000 or less, 1,4-glucan (C) is characterized by containing soluble β-1,3-1,4-glucan having a molecular weight of 500 or more and 20,000 or less.

Patent Document [0009] The present invention relates to a β-glucan-containing grain carbohydrate and a process for producing the same, wherein a grain containing β-glucan is used as a raw material and a protease having a β-glucanase activity of 0 to 10% , Liquefying enzyme having a? -Glucanase activity of 0 to 10% and starcholytic enzyme having a? -Glucanase activity of 0 to 10%, followed by solid-liquid separation, And 1 to 10% by mass of? -Glucan obtained by spray drying or freeze-drying.

In the above Patent Documents 0001 to 0009, β-glucan can be obtained from cereals such as barley but there is a problem that the separation rate of β-glucan from the barley cell wall is not high and there is a limit in obtaining low molecular weight β-glucan.

(0001) KR 1999-0065617 (August 5, 1999) (0002) KR 1489601 (2015.01.28) (0003) KR 0442149 (July 19, 2004) (0004) JP 2002-097203 (2002.04.02) (0005) JP 1994-083652 (Oct. 26, 1994) (0006) JP 2001-323001 (2001.11.20) (0007) JP 2005-307150 (November 4, 2005) (0008) JP 2009-263655 (Nov. 12, 2009) (0009) JP 2009-050226 (2009.03.12)

In order to solve the problems of the prior art, it is an object of the present invention to provide a method for extracting low molecular weight? -Glucan from barley capable of effectively separating and extracting low molecular weight? -Glucan from a barley cell wall.

According to an aspect of the present invention,

a) pulverizing barley to obtain a barley powder;

b) adding hot water to the barley powder and stirring to swell the? -glucan;

c) treating the suspension of the barley powder swollen with the? -glucan with a protease and? -amylase to extract? -glucan. The method of extracting low molecular? -glucan from barley is also provided.

In step b), hot water at 40 to 50 ° C is added to the barley powder and stirred.

In the step c), 3 to 6 parts by weight of protease and 3 to 6 parts by weight of? -Amylase are preferably treated in 100 parts by weight of the barley powder suspension.

In particular, the Lichenase activity of the protease is 0.1 to 0.3 unit / g as a substrate and the Lichenase activity of the α-amylase is 0.1 to 0.2 unit / g as a substrate.

And extracting the above-mentioned? -Glucan and isolating the liquid β-glucan extract by solid-liquid separation.

In addition, the present invention provides a method for separating β-glucan from a liquid phase by solid-liquid separation, α-converting the remaining solid starch-separated material with hot water, and subjecting the starch to α-glycosylation to produce an oligosaccharide transfer liquid The present invention provides a method for producing an oligosaccharide transfer liquid.

The method for extracting low molecular weight? -Glucan from barley of the present invention is characterized in that the? -Glucan contained in the hot barley powder is effectively swollen, and the protease and? -Amylase are added to disperse and dissolve the low molecular? It is possible to extract low molecular β-glucan having a distribution ratio of 90% or more.

In addition, the oligosaccharide transferring solution prepared by the saccharification treatment after the? -Glucan extract is separated into the solid phase and separated into the remaining solid starch separates, and the lactic acid bacteria and Bifidus bacteria are proliferated and the flavor is improved have.

Hereinafter, a method for extracting low molecular β-glucan from the barley of the present invention will be described in detail.

The method for extracting low molecular weight? -Glucan from the barley of the present invention comprises the steps of obtaining barley powder by pulverizing barley, adding hot water to the barley powder and stirring to swell? -Glucan, Treating the suspension with protease and? -Amylase.

First, barley is pulverized by a pulverizer or the like to obtain barley powder. At this time, the barley powder is preferably pulverized to 30 to 100 mesh. When grinding to a size of 30 mesh or less, the separation rate of? -Glucan is not good, and when grinding exceeds 100mesh, the synergistic effect of separation of? -Glucan is insufficient and economical efficiency is poor.

Next, hot water is added to the barley powder and stirred to swell the? -Glucan. In order to effectively swell the? -Glucan contained in the barley powder, hot water at 40 to 50 ° C is preferably added to the barley powder at 4 to 6 times the weight of the barley powder and stirred.

Then, the β-glucan is swollen and the protease and α-amylase are added to the swollen barley powder suspension to disperse and dissolve the β-glucan, and then the β-glucan is extracted while maintaining the water base at 40 to 50 ° C.

At this time, in order to achieve low molecular weight and effective extraction of? -Glucan, 0.4-1.0 parts by weight of protease is added to 100 parts by weight of the barley powder suspension, the amount of Lichenase activity is 0.1-0.3 unit / g as a substrate,? -Amylase is used as a barley powder suspension 100 0.4 to 1.8 parts by weight based on 100 parts by weight of the enzyme is added and the content of Lichenase activity is 0.1 to 0.2 unit / g as a substrate.

Then, the suspension of the barley powder from which the? -Glucan was extracted was quenched, quenched and then subjected to solid-liquid separation to obtain a liquid β-glucan extract. Solid phase separated starch fractions were used for the transfer reaction to oligosaccharides.

The liquid β-glucan extract was separated by solid-liquid separation, and the remaining solid starch-separated material was subjected to hydrolysis in the state of adding hot water to the separated starch, and the oligosaccharide transfer liquid was prepared by the sugar transfer reaction.

Specifically, the starch-separated material was heated to 80 to 90 캜 for 15 minutes by adding warm water and adjusted to a solid content concentration of 30%, and then subjected to α-sterilization, followed by cooling to 50 캜. Amylase was added to the cooled adjusting solution in an amount within the acceptable range of Lichenase activity to leave insoluble dietary fiber and subjected to saccharification treatment at 55 ° C for 60 to 120 minutes and heat treatment at 85 to 90 ° C for 15 minutes to obtain α- Remove the vitality of amylase and cool to 55 ℃.

Amylase and transglucosidase were added to the solution after completion of the glycation reaction, and the sugar transfer reaction was carried out at 55 ° C for 120 to 360 minutes while stirring. From the measurement of the maltose content, Lt; 0 > C for 10 minutes to remove the energetic activity of the enzyme, followed by cooling to below 10 DEG C to obtain oligosaccharide.

Hereinafter, a method for extracting low molecular weight? -Glucan from the barley of the present invention will be described in detail as follows.

[ Example  One; [beta] -glucan extraction]

Barley (varieties; pickle barley) were pulverized using a pulverizer to obtain barley powder of 30 to 100 mesh. Then, 20 g of the barley powder and 80 ml of hot water at 40 to 50 캜 were added to a 200 ml Erlenmeyer flask and dispersed with stirring. Then, 0.8 g of Papain F and 0.4 g of Sumizyme L were added and dispersed and dissolved. The mixture was allowed to stand for 120 minutes in a water base at 45 to 55 ° C to extract β-glucan and cooled to 10 ° C.

Then, a liquid β-glucan extract was obtained by solid-liquid separation by vacuum suction filtration using a polyethylene filter cloth.

On the other hand, Papain F was used with 0.1 to 0.3 unit / g of Lichenase activity as a substrate and Sumizyme L with 0.1 to 0.2 unit / g of Lichenase activity as a substrate.

Measurement of? -glucan content and molecular weight distribution

The content of β-glucan in the liquid β-glucan extract was measured by solid-liquid separation, and a part of the extract was purified and used for measurement of the molecular weight. As a result, the molecular weight distribution of β-glucan was as shown in Table 1 .

At this time, 0.05 gEq / ml.in H ₂ O of the purified β-glucan solution was filtered on No. 5C filter paper, and the filtrate at 0.45 μm was used as a molecular weight measurement sample.

 The column was filled with 1 μl of sample in HPLC apparatus using a Sugar KS-805, MillQ water, flow rate of 1.0 ml / min, temperature of 70 ° C and detector Corona. Pullulan standard samples were used to estimate the molecular weight.

P5: Glucose, P-5. 10, 50, 200, and 800 were used as control samples at 1 mg / ml. In H ₂ O, respectively.

Molecular weight of? -glucan Distribution ratio 80 × 10 ⁴ or more 0.7% 80 × 10 ⁴ to 60 × 10 ⁴ 6.4% 60 x 10 ⁴ to 50 x 10 ⁴ 51.8% 50 × 10 ⁴ to 40 × 10 ⁴ 20.8% 40 x 10 ⁴ to 20 x 10 ⁴ 3.3% 20 x 10 ⁴ or less 16.9%

As can be seen in Table 1, the molecular weight distribution of the extracted? -Glucan is 0.7% in the molecular weight of 800,000 or more, while 99.3% of the molecular weight of 200,000 to 800,000 is most distributed, and particularly 92.8% It can be seen that most of them are occupied.

[ Example  2; Oligosaccharide conversion]

To 60 g of solid starch separated by solid-liquid separation in Example 1, 140 ml of hot water was added to adjust the concentration of the residue to 30%, and the mixture was heated at 80 to 90 ° C for 15 minutes to α -vertize and sterilize the starch Lt; 0 > C. Then, 2.0 g of Sumizyme L was added to the α-starch solution and saccharification reaction was carried out for 60 to 120 minutes. The mixture was heated at 80 to 90 ° C. for 15 minutes and then cooled.

After completion of glycation, 0.6 g of β-amylase # 1500 and 0.3 g of Transglucosidase L-500 were added, and the mixture was allowed to stand at 55 ° C. for 5 hours and then sterilized and cooled at 85 to 90 ° C. for 15 minutes to obtain an oligosaccharide transfer liquid.

Lactobacillus, Bifidus  Culture test

Supplying insoluble dietary fiber, water-soluble dietary fiber and oligosaccharide to humans is important to maintain the balance of good bacteria and bad bacteria alive in the intestines, and is important for maintaining balance of good bacteria such as Bifidus bacteria. Therefore, a culture test was carried out to confirm the effect of the oligosaccharide transfer liquid of Example 2 on lactic acid bacteria and Bifidus bacteria.

Commercial milk was used as a raw material for the fermentation test. LPBA (manufacturer: Proquiga / Spain) was used as a kind of lactic acid bacteria. Fermentation was carried out at 37 ° C for 7 hours and immediately after fermentation, the product was cooled. Milk, oligosaccharide transferring solution, 10% lactose solution and lactic acid bacteria culture were fermented in a mixing ratio (volume ratio) as shown in Table 2.

A B C D E milk 100 80 60 80 60 Oligosaccharide transfer liquid - - - 20 40 10% lactose solution - 20 40 - - Lactic acid bacteria culture solution 3 3 3 3 3

The number of lactic acid bacteria was measured by the BCP medium for the fermentation broths A to E of Table 2, and the number of Bifidus bacteria was measured by the BL medium. The sensory evaluation on the flavor was performed by five adult panelists The results are shown in Table 3.

A B C D E PH 4.3 4.3 4.3 4.3 4.3 Number of lactic acid bacteria 2 × 10 ⁹ 1 x 10 ⁹ 1 x 10 ⁹ 2 × 10 ⁹ 3 × 10 ⁹ Bifidus bacteria 1 x 10 ⁸ 5 × 10 ⁷ 5 × 10 ⁷ 1 x 10 ⁸ 2 x 10 ⁸ zest 23 20 19 25 27

As can be seen in Table 3, the fermentation broths D and E containing the oligosaccharide transferring solution were found to be superior in the number of lactic acid bacteria and flavor to A, B and C, respectively. Since barley? -Glucan is not digested in the human body but is fermented and decomposed by bacteria in the intestine after reaching the intestines, the effect on the intestinal bacterial group is significant, and the growth of Bifidus, which is the most beneficial bacteria in human body, It is considered that it has a considerable influence.

Claims (8)

a) pulverizing the barley to obtain a barley powder;
b) adding hot water to the barley powder and stirring to swell the? -glucan;
c) extracting β-glucan from the barley by treating the β-glucan swelled barley powder suspension with protease and α-amylase to extract β-glucan.
The method according to claim 1,
Wherein the step b) comprises adding hot water at 40 to 50 캜 to the barley powder and stirring the resultant. The method of extracting low molecular β-glucan from barley.
3. The method of claim 2,
Wherein the step c) comprises treating 3 to 6 parts by weight of protease and 3 to 6 parts by weight of? -Amylase in 100 parts by weight of the barley powder suspension, and extracting the low molecular weight? -Glucan from the barley.
The method of claim 3,
Wherein the Lichenase activity of the protease is 0.1 to 0.3 unit / g as a substrate.
5. The method of claim 4,
Wherein the Lichenase activity of the? -Amylase is 0.1 to 0.2 unit / g as a substrate. A method for extracting low molecular? -Glucan from barley.
6. The method of claim 5,
Glucan, and extracting the β-glucan from the liquid phase by solid-liquid separation after extracting the β-glucan, and extracting the low-molecular β-glucan from the barley.
[Claim 6] The method according to claim 6, wherein the liquid β-glucan extract is separated by solid-liquid separation, and the remaining solid starch separation product is subjected to glycosylation by adding hot water to the separated starch product, and the oligosaccharide transfer liquid is produced by the sugar transfer reaction Of the oligosaccharide transfer liquid.
An oligosaccharide transfer liquid characterized by being produced according to claim 7.