JP3425664B2 - Polysaccharide food material promoting calcium absorption and method for producing the same - Google Patents

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a food material containing a polysaccharide that promotes calcium absorption as a main component and a method for producing the same. More specifically, it relates to a water-soluble phosphorylated polysaccharide that promotes calcium absorption in the intestinal tract and a method for producing the same, and an object thereof is to provide a novel polysaccharide that promotes absorption of calcium in foods.

[0002]

2. Description of the Related Art In recent years, it has been said that insufficient intake of calcium causes various diseases such as osteoporosis and increased number of bone fractures. Therefore, in addition to mineral-fortified products to which calcium salts have been added as food components, research has been conducted on components that promote absorption of calcium.

As a component that promotes calcium absorption, a casein phosphopeptide (hereinafter abbreviated as CPP), which is a trypsin decomposition product of casein, is disclosed in Japanese Patent Laid-Open No. 05-26888.
3, JP-A-07-241172), partially decomposed phytic acid (JP-A-04-270296), lactosucrose (JP-A-04-349868), theanderose (JP-A-04-360664), and casein calcium decomposition product (JP-A-05). -219898), lactobionic acid (JP-A-07-277991), xylooligosaccharide (JP-A-07-277).
-67575), a chitin-containing substance (Japanese Patent Laid-Open No. 09-6725).
7), heat-coagulable glucan (JP-A-10-15817)
3) etc. have been reported.

Oligosaccharides (Starch Science Vol. 37 No. 2
Page 76 (1990) and dietary fiber (Dietary Fiber, Japan Dietetic Society, Daiichi Shuppan Co., Ltd., p. 6 1995) are not decomposed by various digestive enzymes, so they reach the intestinal tract as they are and are decomposed by intestinal microorganisms. It is reported that absorption of calcium is promoted by the action of short-chain fatty acids such as acetic acid produced (American Society
for Clinical Nutrition Volume 63 574-578
Page 1996 and Bioscience, Biotechnology
and Biochemistry Volume 59 Issue 2 236-239
P. 1995). On the other hand, CPP prevents the formation of an insoluble precipitate as calcium phosphate in an aqueous solution because the aggregate of phosphorylated serine in the molecule forms a soluble complex with calcium, and the absorption rate in the intestinal tract. Are reported to be increasing (Agricaltual and Biolog
ical Chemistry Vol. 43, 2009-2011, page 1
979).

Thus, oligosaccharides, dietary fiber and CP
Although P has been studied and commercialized, up to now, there has been no component which has both a calcium solubilizing ability and a calcium absorption promoting function due to the formation of short chain fatty acids.

[0006]

DISCLOSURE OF THE INVENTION The present invention prevents the formation of insoluble precipitates of calcium in the intestine,
It is to increase the absorption rate of calcium in the living body and to manufacture food materials having that function. Therefore, a molecular assembly of phosphate groups is formed in the molecules of dietary fiber-like substances (water-soluble polysaccharides), the action of phosphate groups prevents precipitation as insoluble calcium phosphate in the intestine, and short-chain due to colon fermentation. The purpose was to increase the absorption rate of calcium in the intestine by producing fatty acids.

The conventional method for introducing a phosphate group into sugars or polysaccharides is to directly esterify cellulose or other polysaccharides with an organic acid or a salt thereof in an organic solvent in the presence of sulfonyl chloride ( JP-A-01-2
49801), synthesis of sugar 1-phosphate from 4-pentenyloxyglycoside by Fraser-Reid reaction (Journal
of Organic Chemistry Volume 56 Issue 14 4547
-4549 (1991), synthesis of ester by aldol condensation reaction using aldolase which is a glycolytic enzyme (Supplement of Chemistry, Vol. 122, pp. 31-38, 1992).
, Soy protein phosphorylation (Journal of Food Sc
ience, Vol. 48, pp. 716-721, 1983).

As described above, most of the phosphoric acid groups are introduced into the polysaccharide by using an insoluble polysaccharide such as cellulose, and many by using an organic solvent as a reaction solvent, the water-soluble polysaccharide as it is. Cannot be applied to. Furthermore, according to the study of CPP, in order to inhibit the precipitation formation of calcium salt in the small intestine, the phosphate group alone does not have the solubilizing ability, and multiple phosphate groups must be present in the molecule. It is also difficult to use the method of introducing a phosphate group into the. On the other hand, the protein phosphorylation method cannot be used as it is because the shape of the molecule is greatly different.

[0009]

Then, as a result of intensive research, the inventors of the present invention aimed at boiling water-soluble polysaccharide in an aqueous solvent once and then reacting inorganic phosphate under alkaline conditions. Succeeded in obtaining water-soluble phosphate polysaccharide,
It was confirmed that the obtained water-soluble phosphorylated polysaccharide inhibits the precipitation of calcium salt in vitro and promotes the absorption of calcium in the intestine.

The water-soluble polysaccharides of the present invention include water-soluble pectins such as galacturonan, plant gums such as galactomannan, viscous substances such as glucomannan, and synthetic water-soluble polysaccharides such as polydextrose. Any substance can be used as long as it is a polysaccharide that is soluble in water.

In the production method according to the present invention, the water-soluble polysaccharide is dissolved in water, the solution is boiled, then the inorganic phosphate is dissolved, the pH is made alkaline, and the reaction is carried out at a constant temperature of 40 to 50 ° C. Let After completion of the reaction, the reaction solution is purified by dialysis, and the reaction purified solution is powdered by a freeze-drying method to obtain the target phosphorylated polysaccharide.

The inorganic phosphate used is sodium trimetaphosphate, and the amount used is preferably 1.5 per 1 water-soluble polysaccharide, although it depends on the type of water-soluble polysaccharide. Although the amount of the phosphate ester contained in the target substance obtained by increasing the amount of the inorganic phosphate increases, it already shows sufficient calcium salt precipitation formation inhibiting ability at 1.5,
From the viewpoint of cost, the addition amount is set as described above.

The optimum pH of the reaction solution is preferably alkaline conditions. This is because the reaction between sodium trimetaphosphate and the hydroxyl group of the water-soluble polysaccharide proceeds only under alkaline conditions.

The reaction temperature is preferably around 45 ° C. In this manufacturing method, it is necessary to activate the movement of molecules to some extent,
Below this temperature, sufficient acceleration of the reaction cannot be expected, but
Even if the temperature is raised higher than this, it is not possible to expect the promotion of the reaction commensurate with the energy consumption.

The reaction time is preferably 4 hours. This is because the reaction does not proceed sufficiently below this value and the reaction is completed above this value, and there is no great difference in the results obtained with the reaction product for 4 hours.

After completion of the reaction, dialysis is performed to purify the reaction substance. In addition to the target substance, unreacted metaphosphoric acid, reaction by-product pyrophosphoric acid, and the like coexist in the reaction solution, and these must be removed. Since their molecular weight is smaller than that of water-soluble polysaccharides, the target substance is purified using a dialysis membrane. For this reason, if the water-soluble polysaccharide is too small, it becomes difficult to select a dialysis membrane and it takes time for dialysis. In the case of a polysaccharide having a molecular weight of about 10,000, it can be almost purified in about 24 hours.

After the purified reaction product is frozen, it is dried and powdered by a freeze dryer.

The amount of phosphoric acid in the water-soluble phosphoric acid polysaccharide, which was the purified product of the reaction, was determined by using the molybdenum blue colorimetric method.

When the water-soluble phosphorylated polysaccharide obtained by the present invention is ingested with food, it is possible to promote calcium absorption.

[0020]

BEST MODE FOR CARRYING OUT THE INVENTION The present invention applies the principle of transesterification reaction in an alkaline solution. Not only water-soluble polysaccharides but also hydroxyl groups that release a proton in an alkaline solution are incorporated into the molecule. It is considered possible to broaden the application to macromolecules (including peptides) that have.

In the present invention, the water-soluble phosphorylated polysaccharide thus obtained can be used alone as a calcium absorption promoter for tablets, granules, capsules and the like. In addition, calcium absorption can be promoted by adding it to various drinks such as soft drinks and fermented drinks, dairy products, breads and candies. Furthermore, the absorption of calcium can be promoted by adding it to calcium-free foods together with calcium. Concerning the amount of intake, since water-soluble polysaccharides may induce diarrhea when ingested in large amounts, it is preferably about 5% with respect to the amount of food intake.

[0022]

EXAMPLES The present invention will be described in more detail below with reference to Examples and Test Examples, but the present invention is not limited thereto.

[0023]

Example 1 Preparation of water-soluble phosphorylated polysaccharide (1) 10 g of guar gum was dissolved in 100 ml of water and heated to 5
Boil for a minute. 15 g of sodium trimetaphosphate was dissolved in this solution, and the pH was adjusted to 12.5 with 40% sodium hydroxide. During the subsequent reaction, the pH was adjusted to 12.5 by the titration method. The reaction temperature is 45
While maintaining the temperature at 0 ° C., the mixture was stirred with a stirrer and reacted for 4 hours. After completion of the reaction, a dialyzed membrane having a molecular weight cut off of 8,000 (Spectrapore, Spectrum) was dialyzed in running water for 24 hours to obtain phosphorylated guar gum.

[0024]

[Test Example 1] Confirmation of the inhibitory effect on the formation of calcium phosphate precipitate (1) Example 1 was added to 50 ml of 20 mM phosphate buffer of pH 7 and 8.
The phosphorylated guar gum obtained in step 10, 20, 50, 10
Add 0 mg each, add calcium chloride to a final concentration of 5 mM, and leave at 37 ° C for 1 hour,
The mixture was centrifuged at 3,000 rpm for 10 minutes, the supernatant was diluted 100 times and quantified with an atomic absorption spectrophotometer (Z-6100, manufactured by Hitachi, Ltd.), and the calcium solubilization rate was calculated by the formula shown below. Calcium solubilization rate (%) = (calcium concentration in supernatant / calcium concentration in buffer) × 100 As shown in FIG. 1 of the attached sheet, guar gum before phosphorylation has almost the same calcium solubilizing ability. Although not shown, phosphorylated guar gum showed a high calcium solubilizing ability.

[0025]

Test Example 2 Calcium Absorption-Promoting Action in Rats 24 SD male rats with an initial body weight of about 300 g and 9 weeks of age were placed in a metabolic cage and preliminarily reared for 1 week. , The phosphorylated guar gum-added food group and the guar gum-added food group were each divided into 6 groups and subjected to test breeding for 2 weeks. During the test breeding, the diet (Ca: 4.5 g / kg) containing calcium carbonate and a trace amount of calcium phosphate as shown in Attachment 1 and demineralized water as shown in Table 1 of the Appendix and demineralized water were freely ingested, and they were bred in a light-dark cycle every 12 hours. The final 3 days of the 1st week and the 2nd week were collected as the balance test period, and the amount of calcium in the feces was measured with an atomic absorption spectrophotometer (A-6100, manufactured by Shimadzu Corporation). The apparent calcium absorption rate was calculated by the following formula. Apparent calcium absorption rate (%) = [(amount of calcium in the ingested feed) / (amount of chromium in the ingested feed)-
(Amount of calcium excreted in feces) / (Amount of chromium excreted in feces)] / [(Amount of calcium in the ingested feed)
/ (Chromium amount in the ingested feed)] × 100 As shown in Figure 2 of the attached sheet, the results are compared with those in the group in which no polysaccharide was added to the diet (FF) and the group in which guar gum was added,
Calcium absorption was significantly higher in the group to which phosphorylated guar gum was added.

[0026]

Example 2 Preparation of Water-Soluble Phosphorylated Polysaccharide (2) In Example 1, polydextrose (manufactured by Wako Pure Chemical Industries, Ltd.) was used as the water-soluble polysaccharide, and the amount of dissolution was the solubility of polydextrose in water. From the problem of 10g to 4g, 15g of sodium trimetaphosphate according to it
To 6 g, and the dialysis membrane was adjusted to 8,000 to 1,000 based on the molecular weight of polydextrose. That is, 4 g of polydextrose was dissolved in 100 ml of water and boiled for 5 minutes with a gas burner. 3 g of sodium trimetaphosphate are dissolved in this solution and the pH is adjusted to 12.5 with 40% sodium hydroxide. The pH was constantly adjusted to 12.5 during the subsequent reaction. The reaction temperature was 45 ° C. in hot water roasting on a hot stirrer, and the reaction was carried out for 4 hours while stirring with a stirrer. After completion of the reaction, dialysis membrane having a molecular weight cut off of 1,000 (Spectrapore, Spectrum Co.) was dialyzed for one day in running water to obtain phosphorylated polydextrose.

[0027]

Test Example 3 Confirmation of Calcium Phosphate Precipitation Formation Inhibitory Action (2) The polysaccharide added to the phosphate buffer in Test Example 1 was the phosphorylated polydextrose obtained in Example 2. That is, 10 or 2 of the phosphorylated polydextrose obtained in Example 2 was added to 50 ml of 20 mM phosphate buffer of pH 7 and 8.
0, 50, 100 mg was added to each, and calcium chloride was added thereto so that the final concentration was 5 mM, and the mixture was allowed to stand at 37 ° C. for 1 hour, then centrifuged at 3,000 rpm for 10 minutes, and the supernatant was diluted 100-fold. Analysis with an atomic absorption spectrophotometer,
The calcium solubilization rate was calculated by the formula shown below. Calcium solubilization rate (%) = (calcium concentration in supernatant / calcium concentration in buffer) × 100 As shown in FIG. 3 in the attached sheet, the result shows that the calcium content of phosphorylated polydextrose was higher than that of polydextrose. It showed solubilizing ability.

[0028]

Example 3 Preparation of Water-Soluble Phosphorylated Polysaccharide (3) In Example 1, carboxymethylcellulose (manufactured by Wako Pure Chemical Industries, Ltd.) was used as the water-soluble polysaccharide, and the amount of dissolution was the solubility of carboxymethylcellulose in water. From the problem of
0 g was dropped to 2 g, sodium trimetaphosphate was adjusted to 15 g to 1.5 g, and the dialysis membrane was 8,000 to 1,0 depending on the molecular weight of carboxymethyl cellulose.
It was set to 00. That is, 2 g of carboxymethyl cellulose was dissolved in 100 ml of water and boiled for 5 minutes with a gas burner. 1.5 g of sodium trimetaphosphate is dissolved in this solution and the pH is adjusted to 12.5 with 40% sodium hydroxide. The pH was constantly adjusted to 12.5 during the subsequent reaction. The reaction temperature was 45 ° C. in hot water roasting on a hot stirrer, and the reaction was carried out for 4 hours while stirring with a stirrer. After the reaction, the molecular weight cutoff 1,
It was dialyzed for one day in running water using 000 dialysis membrane to obtain phosphorylated carboxymethyl cellulose.

[0029]

Test Example 4 Confirmation of Calcium Phosphate Precipitation Formation Inhibitory Action (2) The polysaccharide added to the phosphate buffer in Test Example 1 was the phosphorylated carboxymethyl cellulose obtained in Example 3. That is, 50m of 20mM phosphate buffer of pH 7 and 8
10, 20, 50, 100 mg each of the phosphorylated carboxymethyl cellulose obtained in Example 3 was added to 1, and calcium chloride was added thereto to a final concentration of 5 mM, and the mixture was allowed to stand at 37 ° C. for 1 hour. 000 rpm,
After centrifugation for 10 minutes, the supernatant was diluted 100-fold and analyzed with an atomic absorption spectrophotometer, and the calcium solubilization rate was calculated by the following formula. Calcium solubilization rate (%) = (calcium concentration in supernatant / calcium concentration in buffer) × 100 The result is that carboxymethylcellulose originally had some calcium solubilizing ability as shown in FIG. Showed higher calcium-solubilizing ability by phosphorylation.

[0030]

The water-soluble phosphorylated polysaccharide according to the present invention is
By the effect of the phosphate group introduced into the molecule, the precipitation formation of calcium phosphate is inhibited in the small intestine, and the absorption of calcium can be promoted by keeping the orally ingested calcium in a solubilized state that is easily absorbed. In addition, as an added value, polysaccharides are decomposed into short-chain fatty acids by the action of intestinal microorganisms in the cecum, so that the action can be expected to enhance calcium absorption in the large intestine, intestinal action, and prevent colon cancer.

[Brief description of drawings]

[Table 1] 3 is a table of dietary components for raising rats used in Test Example 2 of Example 1 according to the present invention.

FIG. 1 is a result of a calcium phosphate precipitation formation inhibitory effect in Test Example 1 of Example 1 according to the present invention.

FIG. 2 shows the results of the calcium absorption promoting action in rats in Test Example 2 of Example 1 according to the present invention.

FIG. 3 is a result of the inhibitory effect on the formation of calcium phosphate precipitate in Test Example 3 of Example 2 according to the present invention.

FIG. 4 is a result of a calcium phosphate precipitation formation inhibitory effect in Test Example 4 of Example 3 according to the present invention.

   ─────────────────────────────────────────────────── ─── Continued front page    (72) Inventor Takanori Kasai               North Sea, Kita 9-jo Nishi 9-chome, Kita-ku, Sapporo-shi, Hokkaido               Hokkaido University Faculty of Agriculture (72) Inventor Yuzo Asano               Hokkaido, Ebetsu City Bunkyodai Midoricho 589-4 North Sea               Provincial Food Processing Research Center                (56) Reference JP-A-8-104696 (JP, A)                 JP-A-56-86901 (JP, A)                 Japanese Patent Publication Sho 44-16387 (JP, B1)

Claims (2)

(57) [Claims] 1. A phosphoric acid characterized in that a water-soluble polysaccharide is dissolved in water, an aqueous solution is boiled, and then sodium trimetaphosphate is dissolved in the solution to adjust the pH to an alkaline side and reacted at a temperature of 40 to 50 ° C. Method for producing polysaccharides 2. The method for producing a phosphorylated polysaccharide according to claim 1, wherein the water-soluble polysaccharide is guar gum.