JPH10262570A - Iron-caseinphosphopeptide complex and production thereof - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain the subject complex exhibiting specific properties, and useful for the prevention and treatment of anemia, and as a raw material of a food, a beverage, a medicine and a feed as an iron reinforcement. SOLUTION: This method for producing an iron-caseinphosphopeptide complex is to mix a solution containing carbonic acid and/or bicarbonic acid with a solution containing a caseinphophopeptide and a solution containing iron for obtaining a carbonic acid and/or bicarbonic acid-iron-caseinphosphopeptide complex showing properties of (A) containing 1-50 atom iron base on 1 molecule of the caseinphosphopeptide, (B) dissolving >=5 wt.% complex in deionized water at 20 deg.C, (C) not forming a precipitation by heating an aqueous solution thereof at 90 deg.C for 10 min and (d) not showing an astringent taste characteristic to iron.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a carbonate and / or bicarbonate-iron-casein phosphopeptide complex. The present invention also relates to a method for producing a carbonate and / or bicarbonate-iron-casein phosphopeptide complex. The carbonic acid and / or bicarbonate-iron-casein phosphopeptide complex of the present invention is characterized by having good solubility, heat resistance, and not exhibiting iron-specific astringent taste.
It is useful as a raw material for foods and drinks, pharmaceuticals, feeds, etc. for the purpose of preventing or treating anemia and fortifying iron.

[0002]

2. Description of the Related Art Since 1975, the iron intake of Japanese people has been flat at around 100% of the required amount, and iron is a dietary nutrient that must be taken with care. One can say. Also, globally, iron is often considered a nutrient deficiency, especially in the supply of iron-enriched foods and medicines for infants under one year of age, athletes, people with anemia, and expectant mothers. Is desired. However, iron salts such as iron sulfate and iron citrate, which are generally used for iron fortification, may be added to foods and drinks due to the problem of feeling the astringent taste unique to iron and damaging the gastrointestinal mucosa. There is a limit. Heme iron, which is an organic iron, also has problems in flavor such as metallic taste and fishy smell, and its addition to foods and drinks has many restrictions.

For the purpose of promoting iron absorption, addition of lactoferrin (JP-A-63-22525) and addition of casein phosphopeptide (JP-A-59-1628) have been proposed.
No. 43) has been tried, but there is a problem that when these substances are mixed with a large amount of iron salt, a precipitate is formed.
It could not be used as a raw material for water-soluble foods and drinks and pharmaceuticals.

[0004]

SUMMARY OF THE INVENTION The present inventors have previously described
By combining iron and casein, an iron agent having weakened the astringent taste peculiar to iron was developed (JP-A-2-83400).
However, iron casein in which iron and casein are combined has poor heat resistance, and is sterilized by heating at 90 ° C. for 10 minutes and at 120 ° C.
When heat treatment such as heat sterilization for 3 seconds or retort sterilization is performed, it has a drawback that it exhibits an astringent taste unique to iron. It is considered that this defect is due to the fact that the bond between iron and casein is weak, so that iron is released from casein by heating and iron hydroxide or the like is generated.

Accordingly, the present inventors have further studied and
They have found that the use of carbonic acid and / or bicarbonate can strengthen the binding between iron and casein, and succeeded in obtaining a carbonic acid and / or bicarbonate-iron-casein complex (Japanese Patent Application No. Hei 7 (1994) -107). -259572). Since the carbonate and / or bicarbonate-iron-casein complex has heat resistance and does not exhibit an astringent taste peculiar to iron even when sterilized by heating, it is useful for preventing or treating anemia. It is useful as a raw material for foods and drinks, pharmaceuticals, feeds, etc. for the purpose of iron fortification.

The inventors of the present invention have a feature that they have heat resistance, do not exhibit the unique astringent taste of iron even when sterilized by heating, and have a more soluble iron-binding property. In order to obtain the substance, we have been conducting intensive research. By mixing carbonic acid and / or bicarbonate, iron and casein phosphopeptide, it has good solubility, heat resistance, and a unique astringent taste of iron. The present inventors have found that a carbonate and / or bicarbonate-iron-casein phosphopeptide complex having the characteristic of not exhibiting the following can be obtained, and have completed the present invention. Accordingly, the present invention provides a novel carbonic acid and / or bicarbonate-iron-casein phosphosol having good solubility, particularly water solubility, having heat resistance and not exhibiting the unique astringent taste of iron. An object is to provide a peptide complex. Another object of the present invention is to provide a method for producing the novel carbonate and / or bicarbonate-iron-casein phosphopeptide complex.

[0007]

DISCLOSURE OF THE INVENTION The carbonic acid and / or bicarbonate-iron-casein phosphopeptide of the present invention has good solubility in water, has heat resistance, and does not exhibit the unique astringent taste of iron. The complex can be i) carbonate, ii) bicarbonate, or iii)
A solution containing carbonic acid and bicarbonate (A solution), iv) a solution containing casein phosphopeptide (B solution), and v) a solution containing iron (C solution) are mixed, and these components are reacted. Obtainable. Alternatively, the reaction product may be concentrated with an ultrafiltration membrane, desalted, and lyophilized to a powder.

[0008] The carbonate and / or bicarbonate-iron-casein phosphopeptide complex thus obtained is as follows:
4). 1) Each casein phosphopeptide contains 1-50 atoms of iron per molecule. 2) 5% (weight) or more dissolved in deionized water at 20 ° C. 3) Heating of an aqueous solution at 90 ° C for 10 minutes. 4) No astringent taste unique to iron. The casein phosphopeptide referred to herein is obtained by decomposing casein with a protease such as trypsin or a protease containing trypsin (for example, pancreatin) and collecting a casein phosphopeptide fraction. Peptide with a molecular weight of about 2,500-4,500 (British Journal of Nutrition, vol. 43, pp. 457-
467, 1980). Alternatively, a commercially available phosphopeptide can be used.

The carbonic acid and / or bicarbonate of the present invention will be described below.
The test example performed in order to investigate the property of an iron-casein phosphopeptide complex is shown.

Test Example 1 (A1 solution) 1 L of aqueous solution containing 100 g of sodium bicarbonate (A2 solution) 1 L of deionized water (B solution) 0.8 L of aqueous solution containing 4 g of casein phosphopeptide (manufactured by Meiji Seika) (C solution) Second chloride Aqueous solution 0.2L containing iron 1.4g D1 solution (1.8L) which mixed A1 solution (1L) and B solution (0.8L)
Of the present invention (bicarbonate-iron-casein phosphopeptide complex) prepared by adding C solution (0.2 L) to A2 solution (1 L) and B
A control product prepared by adding the C solution (0.2 L) to the D2 solution (1.8 L) mixed with the solution (0.8 L) was subjected to heat sterilization (I) at 90 ° C. for 10 minutes. In addition, for the product of the present invention,
With a dialysis membrane of 1,000 cuts, 0.05 mol imidazole and 0.1 mol
After dialysis for 3 days against a buffer solution imitating a liquid food of pH 7.5 containing 15 mol salt, heat sterilization at 90 ° C for 10 minutes
(II) was performed. Properties (1) immediately after preparation, properties (2) after heat sterilization (I), and properties (3) after dialysis and heat sterilization (II) were observed. Table 1 shows the results. According to this, the control product in which casein phosphopeptide and iron alone were mixed resulted in precipitation by heating, but the product of the present invention, in which a complex was formed in the presence of bicarbonate, did not precipitate even when heated. I understand.

[0010]

[Table 1] 試 料 Sample properties (1) Properties (2) Properties (3) ── ───────────────────────────── Inventive product Transparent orange Transparent orange Transparent orange Control product Brown precipitate Brown precipitate ─ ── and and colorless and clear supernatant Colorless and clear supernatant ───────────────────────────────

[0011]

[Test Example 2] The product of the present invention and the control product prepared in the same manner as shown in Test Example 1 were dialyzed (4 ° C, 1 week) against ultrapure water using a dialysis membrane having a molecular weight cut off of 1,000 to complete , And freeze-dried to obtain powders of the product of the present invention and the control product. A dissolution test was performed using these powders. In the dissolution test, the powder was dissolved in deionized water so as to have the respective concentrations, and then the volume was increased to 20 ml, and the mixture was stirred well, centrifuged (3,000 rpm), and the presence or absence of a precipitate was observed. Table 2 shows the results. According to this, it is found that the control product in which casein phosphopeptide and iron alone are mixed has poor solubility, whereas the product of the present invention coexisting with bicarbonate has good solubility.

[0012]

[Table 2] 試 料 Sample concentration Precipitation ─────────────────── Product of the present invention 0.1 (%) None 1 None 5 None 15 Slightly available ─────────────────── Control product 0.1 (%) Available 1 Yes 5 Available 15 Available ───── ──────────────

[0013]

Test Example 3 A solution prepared by dissolving a lyophilized powder of the product of the present invention prepared in the same manner as shown in Test Example 1 in a 50 mM Tris-HCl buffer (pH 9.0) containing 0.1 M sodium chloride to a concentration of 1 mg / ml was prepared. Prepared. As a control, 50 mM of casein phosphopeptide (manufactured by Meiji Seika Co., Ltd.) containing 0.1 M sodium chloride at 1 mg / ml was used.
A solution dissolved in Tris-HCl buffer (pH 9.0) was prepared.
And 50 mM Tris-HCl buffer (pH 9.0) containing 0.1 M salt
After 50 μl of each solution was added to a monoQ HR10 / 10 column (manufactured by Pharmacia) equilibrated in, a linear gradient elution was performed with a 50 mM Tris-HCl buffer (pH 9.0) containing 1 M salt. The flow rate was 2 ml / min, and the detection was carried out by measuring the absorbance at 214 nm. The obtained chart is shown in FIG. 1 and FIG. According to this, it can be seen that a peak not seen in the chart of the control product to which iron is not bound (FIG. 2) exists as the peak A in the chart of the product of the present invention (FIG. 1).

Further, using Q Sepharose FF (manufactured by Pharmacia), a large amount of peak A is collected under the same elution conditions, and the peptide content is measured using a protein assay kit (manufactured by Pierce), and the atomic absorption spectrometer is used. As a result, the iron content was found to be 46% by weight based on the casein phosphopeptide in the complex.

[0015]

BEST MODE FOR CARRYING OUT THE INVENTION The carbonate and / or bicarbonate-iron-casein phosphopeptide complex of the present invention which has good solubility, heat resistance and does not exhibit the unique astringent taste of iron, i) carbonic acid, ii) bicarbonate, or iii) a solution containing carbonic acid and bicarbonate (solution A); iv) a solution containing casein phosphopeptide (solution B); and v) a solution containing iron (solution C). Can be obtained by mixing

In the present invention, the carbonic acid and / or bicarbonate used for producing the carbonic acid and / or bicarbonate-iron-casein phosphopeptide complex may be used in the form of an acid. It may be used in a form. In the present invention, iron used for producing a carbonate and / or bicarbonate-iron-casein phosphopeptide complex is generally used in the form of a water-soluble salt.

In the present invention, when producing a complex of carbonic acid and / or bicarbonate-iron-casein phosphopeptide, i) carbonic acid and / or bicarbonate, ii) casein phosphopeptide, and iii) iron Or i) carbonates and / or bicarbonates in a solid state; ii) casein phosphopeptide; and iii) iron.

In the present invention, the carbonic acid and / or bicarbonate used for producing the carbonic acid and / or bicarbonate-iron-casein phosphopeptide complex includes carbonated water, ammonium bicarbonate, sodium bicarbonate and bicarbonate. Potassium, sodium carbonate, calcium carbonate and the like can be exemplified. Also, as a pH adjuster, sodium hydroxide, ammonia,
Potassium hydroxide, hydrochloric acid, citric acid, lactic acid and the like may be used at the same time. In addition, examples of the water-soluble iron salt include ferric chloride, ferrous sulfate, and iron ammonium citrate.

Further, in the present invention, carbonic acid and / or bicarbonate
The casein phosphopeptide used when producing the iron-casein phosphopeptide complex includes a casein which is obtained by degrading casein with a protease such as trypsin.
Although 2,500 to 4,000 peptides may be used, commercially available casein phosphopeptides may be used.

Hereinafter, the present invention will be described in more detail with reference to Examples.

Example 1 (A solution) 1 L of an aqueous solution containing 100 g of sodium bicarbonate (B solution) 1.6 g of casein phosphopeptide (manufactured by Meiji Seika)
0.8L (C solution) containing 2.8g of ferric chloride hexahydrate 0.2L A solution (1L) and B solution (0.8L) are mixed to prepare D solution (1.8L) The solution C (0.2 L) was added to the solution D (1.8 L) to form a bicarbonate-iron-casein phosphopeptide complex. Next, the solution containing the bicarbonate-iron-casein phosphopeptide complex was concentrated with an ultrafiltration membrane having a molecular weight cutoff of 1,000, and further, ultrapure water was added to concentrate the solution. After desalting to the extent possible, the mixture was freeze-dried to obtain 1.4 g of a bicarbonate-iron-casein phosphopeptide complex.

The peptide content of the bicarbonate-iron-casein phosphopeptide complex was measured using a protein assay kit (manufactured by Pierce), and the iron content was measured using an atomic absorption spectrometer. Contained 11% by weight of iron. Further, when the solution containing the bicarbonate-iron-casein phosphopeptide complex was concentrated with the above-mentioned ultrafiltration membrane, the iron content on the permeate side recovered when the solution was measured by an atomic absorption spectrometer was 1 μg / ml or less. there were.

A sensory evaluation test was conducted on the bicarbonate-iron-casein phosphopeptide complex. That is, panelists of 10 males and 10 females were asked to determine whether or not an aqueous solution in which a bicarbonate-iron-casein phosphopeptide complex was dissolved so as to have a concentration of 50 mg / ml felt an astringent taste. In addition, as a control, the same evaluation was performed for an aqueous solution in which ferric chloride hexahydrate was dissolved so as to have a concentration of 25 mg / ml. Table 3 shows the results. according to this,
It can be seen that the bicarbonate-iron-casein phosphopeptide complex of the present invention does not exhibit any iron-specific astringent taste.

[0023]

[Table 3] 人数 Sample Number of panelists who felt astringent taste ────────────── ───────── Invented product 0/20 Ferric chloride 20/20 ───────────────────────

Further, an animal experiment was conducted on the bicarbonate-iron-casein phosphopeptide complex. That is, a physiological phosphate buffer containing 6.2 mg / 100 g of ascorbic acid and sodium ascorbate as vitamin C (pH
In 7.2), dissolve the bicarbonate-iron-casein phosphopeptide complex of Example 1 so that the iron concentration becomes 20 mg / 100 ml,
For 10 minutes to prepare a sample (the present invention group). On the other hand, a sample using ferrous sulfate instead of the bicarbonate-iron-casein phosphopeptide complex of Example 1 was also prepared (control group 1). Furthermore, a physiological phosphate buffer (pH 7.2) containing 6.2 mg / 100 g of ascorbic acid and sodium ascorbate as vitamin C was heat-sterilized at 90 ° C. for 10 minutes to prepare a sample (control group 2).

From the 21-day-old Wistar female rats (manufactured by Charles River Japan) immediately after weaning, those having a body weight of 45 to 50 g are selected and subjected to iron-free diet (feed having an iron content of 0.25 mg / 100 g: manufactured by Oriental Yeast). Was administered for 2 weeks to produce anemic rats having a blood hemoglobin value of 7 g / 100 mL or less. Each group consisted of 8 to 11 rats. Thereafter, each test sample was administered by oral gavage (sonde) at 1 mL / day for 6 weeks while continuously administering the iron-free diet. Six weeks after the administration of the test sample was started, blood was collected from the tail vein of each rat, and the hemoglobin value was measured using an automatic blood cell counter (manufactured by Toa Medical Electronics). Table 4 shows the results. According to this, it is understood that the bicarbonate-iron-casein phosphopeptide complex of the present invention is more effective in treating anemia than ferrous sulfate used as an iron agent.

[0026]

[Table 4] ──────────────────────── Hemoglobin value (mean ± standard deviation) ───────────── ─────────── Group of the present invention 14.3 ± 0.7 (g / 100mL) Control group 1 12.5 ± 0.8 Control group 2 4.7 ± 0.4 ──────────────── ────────

[0027]

Example 2 (A solution) 1 L of an aqueous solution containing 108 g of sodium bicarbonate (B solution) 50 g of casein phosphopeptide (manufactured by Meiji Seika)
1 L of aqueous solution containing C (solution C) 1 L of aqueous solution containing 84 g of ferric chloride hexahydrate A solution (1 L) and B solution (1 L) are mixed to prepare D solution (2 L), and then D solution (2 L )), Add C solution (1 L) and add bicarbonate-iron-
A casein phosphopeptide complex was generated. And
The solution containing the bicarbonate-iron-casein phosphopeptide complex is concentrated with an ultrafiltration membrane having a molecular weight cutoff of 1,000, and then concentrated by adding ultrapure water until the conductivity reaches 10 μs. After salting and freeze-drying, 46 g of bicarbonate-iron-casein phosphopeptide complex was obtained.

The peptide content of the bicarbonate-iron-casein phosphopeptide complex was measured using a protein assay kit (manufactured by Pierce), and the iron content was measured using an atomic absorption spectrometer. Contained 26% by weight of iron. Further, the bicarbonate-iron-casein phosphopeptide complex was added to water until the concentration became 3% by weight, and the mixture was pasteurized by heating at 90 ° C. for 10 minutes. As a result, no precipitation occurred. Further, the same sensory evaluation test as in Example 1 was performed, but no astringent taste unique to iron was recognized at all.

[0029]

Example 3 Casein phosphopeptide (Meiji Seika) 10
To a solution prepared by dissolving 0 g in 5 L of water was added 250 g of sodium bicarbonate to dissolve the solution. Then, a solution of 164 g of ferrous sulfate heptahydrate dissolved in 5 L of water was added, and a bicarbonate-iron-casein phosphopeptide complex was added. Generated body. Then, the solution containing the bicarbonate-iron-casein phosphopeptide complex was subjected to a molecular weight cut-off of 1,0.
After concentrating to 1 L with an ultrafiltration membrane of 00 and desalting, 1
The mixture was sterilized by heating at 00 ° C. for 5 minutes and spray-dried to obtain 71 g of a bicarbonate-iron-casein phosphopeptide complex.

The peptide content of the bicarbonate-iron-casein phosphopeptide complex was measured using a protein assay kit (manufactured by Pierce), and the iron content was measured using an atomic absorption spectrometer. 13% by weight of iron. When the bicarbonate-iron-casein phosphopeptide complex was added to water to 13% by weight, it was completely dissolved. further,
A sensory evaluation test was conducted in the same manner as in Example 1, but no astringent taste unique to iron was observed.

[0031]

Example 4 To a solution of 120 g of sodium bicarbonate dissolved in 1 L of water was added 50 g of casein phosphopeptide to dissolve the solution. Then, a solution of 100 g of iron ammonium citrate dissolved in 2 L of water was added to the solution. A casein phosphopeptide complex was generated. Then, the solution containing the bicarbonate-iron-casein phosphopeptide complex was concentrated to 1 L with an ultrafiltration membrane having a molecular weight cutoff of 1,000, desalted, lyophilized, and then lyophilized to obtain bicarbonate-iron-casein. 44 g of the phosphopeptide complex was obtained.

The peptide content of the bicarbonate-iron-casein phosphopeptide complex was measured using a protein assay kit (manufactured by Pierce), and the iron content was measured using an atomic absorption spectrometer. Contained 24% by weight of iron. Further, when the bicarbonate-iron-casein phosphopeptide complex was added to water up to 20% by weight, it was completely dissolved. further,
A sensory evaluation test was conducted in the same manner as in Example 1, but no astringent taste unique to iron was observed.

[0033]

[Reference Example 1] After mixing 3 kg of margarine and 2 kg of refined white sugar and whipping, 0.7 kg of whole egg was added and further whipped. And to this whipped thing, flour 10
kg, soy protein 0.2 kg, skim milk powder 0.4 kg, and bicarbonate-iron-casein phosphopeptide complex 1 obtained in Example 4.
A mixture of 3 g was added and mixed, followed by forming and cutting 10 g each. This was baked at 180 ° C. for 11 minutes in a band-type oven to produce a biscuit. One biscuit contained about 2 mg of iron.

[0034]

Reference Example 2 Skim milk powder 6%, butter 1%, bicarbonate-iron-casein phosphopeptide complex obtained in Example 1 0.0
38% and filtered water were added to make the whole 100%, mixed, dissolved and then heat-sterilized to produce an iron-enriched milk beverage. The iron-enriched beverage (100 mL) contained about 10 mg of iron.

[0035]

[Reference Example 3] 33 parts of skim milk powder, 19 parts of flour, 8 glucose
Parts, bread crumbs 7 parts, fish meal 7 parts, defatted soybean meal 4 parts, sugar 4
Parts, vitamins and minerals mixture 3 parts, animal fats and oils 2 parts,
One part of the oligosaccharide and 0.014 part of the bicarbonate-iron-casein phosphopeptide complex obtained in Example 3 were blended to produce a feed for swine. In addition, about 40 mg of iron was contained in 1 kg of the pig feed.

[0036]

Industrial Applicability The carbonate and / or bicarbonate-iron-casein phosphopeptide complex of the present invention has good solubility, heat resistance, and does not exhibit iron-specific astringent taste. It is useful as a raw material for foods and drinks, pharmaceuticals, feeds, etc. for the purpose of preventing or treating anemia and fortifying iron.

[Brief description of the drawings]

FIG. 1 shows an elution chart of the product of the present invention on a monoQ HR10 / 10 column in Test Example 3.

FIG. 2 shows an elution chart of a control product on a monoQ HR10 / 10 column in Test Example 3.

──────────────────────────────────────────────────の Continuation of the front page (51) Int.Cl. ⁶ Identification symbol FI // C07K 103: 00 (72) Inventor Akira Tomizawa 5-3-33 Toyooka, Iruma-shi, Saitama Arden 710 (72) Inventor Hitoshi Aikawa 12-89, Goryo-cho, Higashimatsuyama-shi, Saitama-ken Park-town, 5-5-503 (72) Inventor Ken Takahashi 25-19, Matsubacho, Tokorozawa-shi, Saitama Maison Matsuba 302

Claims (3)

[Claims] 1. Carbonic acid having the following properties 1) to 4) and / or
Or a bicarbonate-iron-casein phosphopeptide complex. 1) Each casein phosphopeptide contains 1-50 atoms of iron per molecule. 2) 5% (weight) or more dissolved in deionized water at 20 ° C. 3) Heating of an aqueous solution at 90 ° C for 10 minutes. 4) No astringent taste unique to iron. 2. A solution containing i) carbonic acid, ii) bicarbonate, or iii) a solution containing carbonic acid and bicarbonate (A solution), iv) a solution containing casein phosphopeptide (B solution), and v) a solution containing iron (C solution) and the following 1) to 4)
Carbonic acid and / or bicarbonate-iron-casein phosphopeptide complex exhibiting the following properties: 1) Each casein phosphopeptide contains 1-50 atoms of iron per molecule. 2) 5% (weight) or more dissolved in deionized water at 20 ° C. 3) Heating of an aqueous solution at 90 ° C for 10 minutes. 4) No astringent taste unique to iron. 3. A solution containing i) carbonic acid, ii) bicarbonate, or iii) a solution containing carbonic acid and bicarbonate (A solution); iv) a solution containing casein phosphopeptide (B solution); and v) a solution containing iron. (C solution), and a method for producing a carbonate and / or bicarbonate-iron-casein phosphopeptide complex exhibiting the following properties 1) to 4). 1) Each casein phosphopeptide contains 1-50 atoms of iron per molecule. 2) 5% (weight) or more dissolved in deionized water at 20 ° C. 3) Heating of an aqueous solution at 90 ° C for 10 minutes. 4) No astringent taste unique to iron.