JP2860489B2 - Food material, bifidobacterium growth promoter and method for producing them - Google Patents

Description

Description: TECHNICAL FIELD The present invention relates to a food material containing a specific trisaccharide as a functional component, a bifidobacterium growth promoter, and a method for producing the same.

In the following description, the trisaccharide compound name “O-α-D-glucopyranosyl- (1 → 6) -O-α” according to the systematic nomenclature is used.
“-D-glucopyranosyl- (1 → 2) β-D-fructofuranoside” is referred to as “Seanderose”.

In the present invention, "%" indicating the blending unit / content means "% by weight" unless otherwise specified.

<Conventional Technology> In recent years, many studies have been conducted on the effects of intestinal bacteria on health, and bifidobacteria (Bifidobacteriu
The growth of m) has been found to be favorable for human health. In other words, the bifidobacteria dominating the intestinal flora is suppression of the production of putrefactive products by putrefactive bacteria,
It is effective in preventing the production of toxic amines and suppressing the growth of pathogenic bacteria.

<Problems to be Solved by the Invention> As a carbohydrate for selectively growing this bifidobacterium,
A bifidobacterium growth activator (proliferation promoter) containing a branched oligosaccharide having an α (1 → 6) bond of glucose molecule (glucose) has been proposed (JP-A-61-227777).

However, the branched oligosaccharides comprising only glucose units such as isomalttriose disclosed in the above publication have relatively low sweetness (see Table 2). In many cases, a combination with a sweetener was necessary.

Also, since isomalttriose is a reducing sugar,
When subjected to heat treatment, the color was easily discolored (see the discoloration degree test described later), and was unsuitable for foods in which color was important.

In addition, when producing from the oligosaccharide raw material, it is desirable that the yield of the functional component is good (especially when used as a bifidobacterium growth promoter). In view of the above, the present invention provides a food material or a bifidobacterium growth promoting agent which has a sufficient function of promoting the growth of bifidobacteria, has a high degree of sweetness, does not easily discolor, and can be expected to be applied over a wide range. The purpose is to:

Another object of the present invention is to provide a method for producing a food material or the like which can obtain the above-mentioned food material or the like with a high content of functional components, that is, in a good yield.

In addition, it is known that seandellose is contained in honey (Advances in Carbohydrate Chemistry, 2
5,285-, 1970) is extremely small. As far as the present inventors know, there is no food material containing Canderylose as a functional ingredient.

<Means for Solving the Problems> In the course of research and development for solving the above problems, the present inventors have found that Ceanderose is well assimilated to bifidobacteria among intestinal bacteria, and E. coli and Clostridium, etc. It is hardly assimilated by harmful bacteria, that is, it has a function of selectively promoting the growth of bifidobacteria (see Table 1), and the sweetness of seandellose is the above α (1 → 6) Two new findings were obtained: higher than oligosaccharides with linkages (see Table 2). Based on these findings, the present inventors have conceived a configuration of the present invention "a food material and a bifidobacterium proliferating agent containing seanderose as a functional component for promoting the growth of bifidobacteria".

Further, the food material of the present invention and the like, in order to obtain a good yield, found that it is sufficient to act a specific enzyme agent,
The production method of the present invention `` A transglycosidase derived from Aspergillus niger is allowed to act as an enzymatic agent on a mixed solution of maltooligosaccharide and sugar to produce a food material or the like containing seanderose as a functional component for promoting the growth of bifidobacteria. Method ".

The food material of the present invention is a food material containing seandellose, and can be in the form of syrup, powder, granule, or the like. At this time, bulking agents, sweeteners, carbohydrates,
Other components such as vitamins and bifidobacteria can be contained, but the content of Canderoose is desirably 1% or more from the viewpoint of its effect.

The bifidobacterium growth promoter of the present invention is a liquid or powdered sweetener, beverages such as soft drinks and milk drinks, breads, candy, candy and the like, various foods and drinks such as health foods, drinks, and tablets. It can be obtained by incorporating seanderose into feeds such as medicines, livestock, and pets. At this time, it is desirable that the content of the seanderose is 1% or more from the viewpoint of the effect.

The method for producing seandellose of the present invention is characterized in that a mixed solution of maltooligosaccharide and sugar is added to a transglucosidase derived from Aspergillus niger (a lower concept of α-glucosidase).
Is made to act.

The maltooligosaccharide used here as a raw material mainly contains maltooligosaccharides such as maltose, maltotriose, and maltotetraose, and includes, for example, those obtained by hydrolyzing starch with an enzyme and / or acid. Among these, hymaltose syrup is particularly industrially advantageous.

As the enzyme agent used in the present invention, an enzyme derived from Aspergillus niger and treated so that transglucosidase has a glucoamylase activity of 1/10 or less is desirable (see Table 3).

A transglucosidase derived from Aspergillus niger is allowed to act on the above mixed solution of maltooligosaccharide and sugar (sucrose), and glucose is transferred to the 6-position of glucopyranose in the sugar to produce seanderose.

At this time, the mixing ratio of starch hydrolyzate and sugar was 1: 100
To 100: 1 are preferred, but 3: 7 to 7: 3 are more preferred. The pH is preferably 4.0 to 6.5, and the temperature is preferably 20 to 70 ° C. Since the amount of the production of Seanderose increases at the beginning and decreases after the equilibrium point, it is desirable to stop the reaction before the amount decreases significantly. As a standard for stopping the reaction, the value of glucose / maltooligosaccharide in the reaction solution is preferably 0.2 to 15 (see Table 4).

The reaction solution obtained above further excludes monosaccharides and / or disaccharides by using a chromatographic separation or a separation membrane with an alkaline earth metal type cation exchange resin column or the like, or mainly contains trisaccharides. Separating and recovering the sections can also increase the content of Chandellose.

In addition, at this time, since Candellose is not easily decomposed by glucoamylase, the above reaction solution is further added to
By reacting glucoamylase to decompose maltriose, panose, and the like, and then performing separation, it is possible to obtain higher-purity seanderose.

The above reaction solution or a product obtained by separating the reaction solution can be decolorized, desalted and purified with activated carbon or an ion-exchange resin, or concentrated to a syrup, and dried into a powder by a spray dryer or the like. it can.

<Effects and Effects of the Invention> The food material and the bifidobacterium growth promoting agent of the present invention are, as shown in the test examples and examples described below, the use of bifidobacterium alone without the growth of bad bacteria in the intestine such as clostridium. Has the effect of selectively proliferating.

The degree of sweetness is higher than that of the conventional trisaccharide having an α (1 → 6) bond, and it is non-reducing.

Therefore, it can be widely applied as a food material or a bifidobacterium growth promoter.

Furthermore, the method for producing a food material or the like of the present invention is characterized in that a mixed solution of maltooligosaccharide and sugar is produced by allowing transglucosidase of Aspergillus niger to act as an enzymatic agent, thereby producing a high content of seanderose, that is,
It is obtained in high yield and is industrially useful.

It is known that raffinose, which is similar to the Candellose of the present invention (having a structure in which a glucosyl group bonded to α (1 → 6) bond is replaced with a galactosyl group), has relatively low selectivity for bifidobacteria. (See the lower left column on page 4 of the aforementioned publication).

<Test Examples and Examples> Hereinafter, test examples and examples performed to confirm the effects of the present invention will be described.

(1) Test for growth activity of bifidobacteria The powder obtained in Example 2 was used as Ceanderose.
For other sugars, assimilation by intestinal bacteria was examined using commercially available reagents.

Medium Used A 10% aqueous solution of a test saccharide, which had been sterilized by filtration, was added to a sterilized Peptone East Fields medium (PYF medium) to a final concentration of 0.5%.

Test Method After culturing the bacteria shown in Table 1 on an agar plate medium, each strain was inoculated into a medium at a concentration of 10 ⁶ CFu / tube and cultured at 37 ° C. for 72 hours under anaerobic conditions.

The absorbance (OD) of this culture at 650 nm was measured,
The growth degree (RG) of each saccharide with respect to glucose was determined by the following equation.

Results The results are shown in Table 1. In Table 1, ++ represents RG> 100 ++, 75 ≦ RG ≦ 100 + represents 50 ≦ RG ≦ 75 ±, 25 ≦ RG <50− represents RG <25.

As is evident from Table 1, seanderose is well assimilated by bifidobacteria among intestinal bacteria and hardly assimilated by harmful bacteria such as Escherichia coli and Clostridium, that is, selectively grows bifidobacteria. It turns out that it has activity.

(2) Sweetness test: Each of the trisaccharides shown in Table 2 was subjected to a sensory test for the sweetness of a 10% aqueous solution at 20 ° C. Sugar (sucrose) was sequentially diluted with tasteless water to obtain a standard sample, and a sensory test was performed by comparing sweetness with the standard sample. The value obtained by dividing the sugar concentration (%) that gives the same sweetness by 10 was taken as the sweetness degree, and the arithmetic average value of the measurement results by 10 panelists was displayed.

From Table 2 showing the results, it can be seen that seandellose has a higher degree of sweetness than any of the other trisaccharides.

Many people commented that the quality of sweetness of seandellose was similar to that of sugar and was of high quality.

(3) Discoloration test for heat treatment: Prepare a 10% aqueous solution (pH 6.0) of each of the Anderose, Sugar, Glucose, and Isomaltriose, add glycine to each to 1%, and add each test solution. After preparation, each test solution was subjected to a heat treatment under the conditions of 120 ° C. for 60 minutes, and the colors before and after heating were compared. The brown intensity was as follows.

Glucose>Isomaltotriose> Sugar = Seanderose.

(4) Test of the degree of influence of each element on the reaction to produce the Anderose: a) Influence of enzymatic agent;
To 0), each of the enzyme preparations in Table 2 was added so that the transglucosidase activity became 300 u, and the mixture was reacted at 60 ° C for 24 hours. At this time, the content of the Candellose was as shown in Table 3.

In addition, the analysis of Seanderose was performed by HPLC.
(Column: Develosil-NH ₂ -5, solvent: CH ₃ CN: water = 3: 1) As a confirmation of this analysis method, peaks of Candellose in HPLC were collected and the following were confirmed.

It is a non-reducing sugar that does not show reducing properties in feeling liquid.

It was decomposed into isomaltose and fructose by 0.1N hydrochloric acid, and the molar ratio was 0.9: 1.0.

It was decomposed into glucose and fructose by 1N hydrochloric acid, and the molar ratio was 2: 1.

Degraded to isomaltose and fructose by yeast cyclase.

From Table 3, it can be seen that the transglucosidase agent is preferably a transglucosidase that has been treated so that the activity of glucoamylase is 1/10 or less.

b) Influence of reaction time; mixed solution of 10 g of maltose and 10 g of sugar (concentration 50%, pH 6.
To 0), a transglucosidase derived from Aspergillus niger (300,000 u / ml, glucoamylase activity = 5000 u / ml, manufactured by Amano Pharmaceutical Co., Ltd.) was added so that the transglucosidase activity became 15000 u, and the mixture was reacted at 60 ° C. The time course of the sugar composition at this time was as shown in Table 4.

From Table 4, it can be seen that the amount of Candellose produced increases at the beginning and decreases after the equilibrium point. Therefore, the reaction was stopped when the value of glucose / maltooligosaccharide in the reaction solution was 0.2 to 15 (more preferably 0.4 to 10). It turns out that the range is preferable.

(5) Example 1. Water was added to high maltose syrup (maltose content = 82%, manufactured by Nippon Cornstarch Co., Ltd.) to make up to 50% by weight. Na
Add OH to adjust the pH to 6.0, and add Aspergillus
1 ml of niger transglucosidase (300000 u / ml, Amano Pharmaceutical Co., Ltd.), glucoamylase activity = 5000 u / ml)
Was added and stirred at 60 ° C. for 24 hours. The reaction solution is decolorized with activated carbon and an ion exchange resin, desalted, purified, concentrated, and solidified.
75% syrup was obtained. This syrup contains 18.6% solids.
% Of seanderose and isomaltooligosaccharides (5.4% isomaltose, 6.9% panose, 1.2% isomaltite triose) and is suitable as a bifidobacterium growth promoter, and at the same time, has a high quality sweetness and sweetness It is also suitable as a material.

(6) Example 2. Water was added to high maltose syrup (maltose content = 82%, manufactured by Nippon Cornstarch Co., Ltd.) to make 50%.
To 1 kg, add 2 kg of a 50% aqueous solution of sugar, mix and add 4% NaOH
To adjust the pH to 6.0, and add transglucosidase from Aspergillus niger (same as in Example 1).
1 ml was added and the mixture was stirred at 60 ° C. for 24 hours. Furthermore, glucoamylase (GNL-3000, manufactured by Amano Pharmaceutical Co., Ltd.) was added to the reaction solution.
1 ml was added and stirred for 6 hours. The content of the Candellose in this reaction solution was 17.8% of the solid content.

Next, 100 g of the reaction solution was applied to a calcium-type cation exchange resin column (SK1B-S, manufactured by Mitsubishi Kasei Corporation, inner diameter 15 cm × 180 c.
Chromatographic separation was carried out using an (mx 4 tower) to separate and collect the trisaccharide-based fraction to obtain a liquid having a solid content of 93.8% of Candellose. This solution is decolorized with activated carbon and ion exchange resin, purified by desalting, concentrated to 70% solids and brought to 20 ° C.
The resulting crystals were collected by standing for 24 hours and washed to obtain 4 g of crystal powder having a content of 99.5% Canderoose per solid content.

This crystal powder is suitable as a bifidobacterium growth promoter and, at the same time, has good sweetness and is also suitable as a powdered sweetener.

(7) Example 3. 1 kg of the syrup of Example 1 was mixed with 3 kg of isomerized sugar having a solid content of 75% to obtain a syrupy sweetener containing 4.7% of seanderose in the solid content.

This is suitable as a food material having a bifidobacterium growth promotion function for various foods such as beverages, confections, and bread.

(8) Example 4 1 kg of the powder of the Anderose and 1 kg of the balatinose powder were mixed to obtain 2 kg of a powdered sweetener. It has a high quality sweetness and is suitable as a food material (sweetener) and a bifidobacterium growth promoter.

──────────────────────────────────────────────────続 き Continuation of the front page (56) References JP-A-60-166693 (JP, A) JP-A-47-20373 (JP, A) (58) Fields investigated (Int. Cl. ⁶ , DB name) A23L 1/22-1/237 A23L 1/24

Claims (4)

(57) [Claims] (1) O-α-D-glucopyranosyl- (1 →
6) -O-α-D-glucopyranosyl- (1 → 2) β-
A food material comprising D-fructofuranoside as a functional component for promoting the growth of bifidobacteria. (2) O-α-D-glucopyranosyl- (1 → 6) -O-α-D- is reacted with a mixed solution of maltooligosaccharide and sugar by using transglucosidase derived from Aspergillus niger as an enzyme agent. Glucopyranosyl- (1 →
2) A method for producing a food material, comprising β-D-fructofuranoside as a functional component for promoting the growth of bifidobacteria. (3) O-α-D-glucopyranosyl- (1 →
6) -O-α-D-glucopyranosyl- (1 → 2) β-
A bifidobacterium growth promoter comprising D-fructofuranoside as a functional component. 4. O-α-D-glucopyranosyl- (1 → 6) -O-α-D- is reacted with a transglucosidase originating from Aspergillus niger as an enzyme agent in a mixed solution of maltooligosaccharide and sugar. Glucopyranosyl- (1 →
2) A method for producing a bifidobacterium growth promoter containing β-D-fructofuranoside as a functional component.