JP3061364B2 - Infant formula - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a novel infant formula having a bone formation promoting effect by blending a milk-derived basic protein and / or a milk-derived basic peptide.

[0002]

2. Description of the Related Art In recent years, the bones of infants and children have become weak, and there has been a problem that bones are easily broken. In addition, the number of patients with various bone diseases such as osteoporosis, bone fracture, and back pain is increasing with the aging. And these causes
Insufficient intake of calcium, lowering of calcium absorbing ability, lack of active vitamin D ₃ secretion, hormonal imbalance, etc. are pointed out. Accordingly, various calcium agents for calcium supplementation have been developed to prevent these various bone diseases. In addition, in order to prevent bone diseases such as osteoporosis, it has become clear that it is effective to increase the bone mass during the growth period including the infant period to increase the maximum bone mass as much as possible. In addition, various calcium agents such as calcium carbonate have been added to infant formula for the purpose of strengthening bones. However, it has been reported that calcium contained in milk is less absorbable than calcium contained in breast milk (Gur
r, MI, J. Dairy Res., vol. 48, pp. 519-554, 1981).

[0003]

DISCLOSURE OF THE INVENTION The present inventors have been searching for a bone growth promoting substance contained in a milk component. As a result, they have found that the basic protein fraction contained in milk has a bone growth promoting effect. And
It has also been found that this basic protein can be easily obtained from milk or raw materials derived from milk by cation exchange resin chromatography and that bone metabolism can be improved by oral ingestion. Furthermore, they found that a basic peptide obtained by hydrolyzing this basic protein with proteolytic enzymes such as pepsin and pancreatin also has a bone growth promoting effect, and proposed a bone strengthening agent containing these substances as active ingredients. (Japanese Patent Application No. 7-207509).

When a diet containing a milk-derived basic protein or a milk-derived basic peptide is administered to rats in the middle to late stages of lactation, bone growth is promoted more than in rats to which a normal diet is administered. This led to the completion of the present invention. Accordingly, an object of the present invention is to provide a novel infant formula having an osteogenesis promoting effect.

[0005]

A feature of the present invention resides in that a basic protein and / or a basic peptide is blended into an infant formula to impart an osteogenesis promoting action to the infant formula.

[0006] The infant formula referred to in the present invention can be broadly classified into infant formula and infant formula. Infant formulas include infant formulas, premature infant formulas, follow-up milks, special formulas for pharmaceuticals, and other infant formulas for infant rearing, and liquid formulas for infants. There is a liquid formula prepared for the purpose of artificial rearing of infants.

The milk-derived basic protein used in the present invention is obtained from mammalian milk such as cow's milk, human milk, goat's milk, sheep's milk or its whey, and has an amino acid composition of lysine and histidine. And 15% by weight or more of basic amino acids such as arginine. Then, this basic protein, for example, after adsorbing the basic protein by contacting a milk material such as skim milk or whey with a cation exchange resin, a salt concentration of 0.1 to 2.0 M, preferably 0.1 to 1.0 M Can be obtained by elution with an eluate of

As a method for obtaining a milk-derived basic protein, a method using an alginate gel (Japanese Patent Application Laid-Open No.
246198), a method using a sulfated product (JP-A-63-255300), a method using inorganic porous particles
(JP-A-1-86839) and a method using a cation exchanger (JP-A-5-202098).
Further, if necessary, the basic protein thus obtained is desalted and concentrated by means of ion exchange, reverse osmosis, ultrafiltration, electrodialysis, etc. Alternatively, it can be used in the form of powder by means such as freeze-drying and spray-drying.

[0009] The milk-derived basic peptide used in the present invention may be obtained by adding pepsin,
It is obtained by the action of proteolytic enzymes such as trypsin, chymotrypsin and pancreatin, and has the same amino acid composition as the basic protein. Then, for example, when the basic peptide is treated with proteolytic enzyme using pepsin, the pH of the basic protein is adjusted to pH 1.5 with an acid such as hydrochloric acid, and after pepsin is acted on, the sodium hydroxide is added. By neutralizing with an alkali such as Further, after a protease such as pepsin is allowed to act, a protease such as pancreatin is further caused to act, and the molecular weight is 2,000 to 20,0.
It is also possible to obtain 00 basic peptides. Further, if necessary, the basic peptide thus obtained is desalted and concentrated by means of ion exchange, reverse osmosis, ultrafiltration, electrodialysis, etc. Alternatively, it can be used in the form of powder by means such as freeze-drying and spray-drying.

[0010]

BEST MODE FOR CARRYING OUT THE INVENTION In the infant formula of the present invention, in the case of a powdered milk, 100-1,000 mg of a milk-derived basic protein and / or a milk-derived basic peptide is blended per 100 g of the product, and In the case of formula milk, it is preferable to mix 13 to 130 mg of milk-derived basic protein and / or milk-derived basic peptide per 100 ml of the product.

[0011] Infant formula is composed mainly of proteins, lipids, carbohydrates, vitamins and minerals, and further contains other active ingredients which can be incorporated into infant formula. May be contained.

The main components of the infant formula are listed below.
Examples of proteins include casein, whey protein concentrate (WPC), whey protein isolate (WPI), αs-
Casein, β-casein, α-lactalbumin, β-
Milk proteins such as lactoglobulin, milk protein fractions, egg proteins, plant proteins such as soy protein and wheat protein can be used. Further, peptides, free amino acids and the like obtained by treating these proteins with proteolytic enzymes or acids may be used as a nitrogen source. Further, amino acids such as taurine, cystine, arginine, glutamine and the like can be used for imparting a specific physiological effect. In addition, these proteins, peptides, and free amino acids are 5 to 5 per solid product.
What is necessary is just to mix so that it may become 30 weight%.

The saccharides include starch, soluble polysaccharide,
Dextrin, sucrose, lactose, maltose, glucose, glucose, galactosyl lactose, fructooligosaccharides, oligosaccharides such as lactulose, artificial sweeteners and the like can be used. These saccharides may be blended so as to be 40 to 80% by weight based on the solid content of the product.

As lipids, animal fats such as milk fat, lard, beef tallow, fish oil, corn oil, soybean oil, rapeseed oil, coconut oil, palm oil, palm kernel oil, safflower oil, perilla oil,
Linseed oil, evening primrose oil, medium chain fatty acid triglyceride (MC
T) and vegetable oils such as cottonseed oil can be used. Further, fractionated oils, hydrogenated oils, and transesterified oils of these fats and oils may be used. These lipids may be blended so as to be 40% by weight or less per solid content of the product.

As the vitamin, vitamin A, vitamin B, vitamin C, vitamin D, vitamin E, vitamin K, folic acid, pantothenic acid, β-carotene, nicotinamide and the like can be used. In addition, these vitamins may be blended so as to be 10 mg to 5 g / 100 g per solid product.

As minerals, calcium, magnesium, potassium, iron, copper, zinc, sodium, iodine,
Manganese, selenium and the like can be used. In addition, these minerals are 1 mg to 5 g / 100 g per solid product.
What is necessary is just to mix so that it may become. For the formulation of vitamins and minerals, refer to the literature (CODEX standards and special hygiene regulations for special-purpose foods including infant foods, CAC / VOL.IX).
-1st edition and Supplement 1,2,3,4, issued by the Japan Dairy Association, 1993;
Edition), Published by Food and Science Company, 1993;
Food Material Natural Products Handbook (12th edition), Published by Food and Science, 19
92) Among the vitamins and minerals listed in
What is necessary is just to select and use what can be used for infant formula.

In preparing the infant formula having the osteogenesis-promoting effect of the present invention, in the case of a powdered milk formula, the solution prepared by blending and homogenizing the above-mentioned ingredients is mixed with a basic protein and / or milk-derived protein. Alternatively, a milk-derived basic peptide is added and mixed, followed by concentration and drying according to a conventional method to obtain a product, or drying of a sterilized milk-derived basic protein and / or milk-derived basic peptide. The powder may be mixed with the infant formula to make a product. In the case of a liquid milk preparation, a sterilized milk-derived basic protein and / or a milk-derived basic peptide are added to a solution prepared by sterilizing the above-mentioned raw materials, mixed, and then cooled according to a conventional method. And make it a product.

Hereinafter, the present invention will be described in detail with reference to Reference Examples, Examples, and Test Examples.

Test Example 1 Using a cation exchange resin (S-Sepharose) column, the concentration of basic protein contained in breast milk, bovine raw milk and commercially available infant formula was measured. That is,
S equilibrated with 50 mM sodium citrate buffer (pH 6.5)
-Each sample was passed through a Sepharose column to adsorb the basic protein. After sufficiently washing the column with deionized water, the basic protein is eluted with a 1 M sodium chloride solution, and the concentration of the protein contained in the eluted fraction is quantified using a protein assay kit (manufactured by BioRad). The concentration of the basic protein was calculated. Table 1 shows the results. In addition, about the commercially available infant formula, the thing melt | dissolved in pure water so that a solid content might be 13% was used.

[0019]

[Table 1]

[0020]

[Reference Example 1] Rennet curdling enzyme in 50,000 liters of raw milk
(Manufactured by Hansen) was added to a final concentration of 0.0032% to form a cheese curd. The whey and curd were separated using a stainless steel mesh to obtain 40,000 liters of unsterilized cheese whey. After passing 40,000 liters of the above-mentioned unsterilized cheese whey through a column filled with 400 kg of a sulfonated chitopearl (manufactured by Fuji Boseki) as a cation exchange resin, the column was sufficiently washed with deionized water. next,
0.02M carbonate buffer (pH 7.0) containing 0.98M sodium chloride
The basic protein fraction adsorbed on the resin was eluted in the above step, desalted and concentrated by a reverse osmosis membrane, and lyophilized to obtain 1.7 kg of a powdery basic protein fraction.

[0021]

Test Example 2 The basic protein fraction obtained in Reference Example 1 was analyzed for its component composition. The protein was quantified by the Kjeldahl method, the fat was quantified by the Soxhlet extraction method, and the ash was quantified by the wet ash method. Table 2 shows the results.

[0022]

[Table 2] ───────────────── Moisture 1.0 (% by weight) Protein 97.0 Fat 0.4 Ash 0.3 Other 1.3 ──

[0023]

Test Example 3 The amino acid composition of the basic protein fraction obtained in Reference Example 1 was analyzed. The amino acid composition was determined by adding the basic protein fraction to a 6N hydrochloric acid solution at 110 ° C and 24 ° C.
After hydrolysis for an hour, amino acid analyzer (Hitachi L-8500)
Was measured. Table 3 shows the results.

[0024]

[Table 3] ア ス Aspartic acid 11 (wt%) Serine 5 Glutamic acid 12 Proline 6 Alanine 6 Leucine 10 Lysine 9 Histidine 3 Arginine 8 Others 30 ─── ───────────────

[0025]

Reference Example 2 A column packed with 4 kg of a cation exchange resin, sulfonated chitopearl (manufactured by Fuji Boseki Co., Ltd.), was loaded with a reference example 1
After passing through 400 liters of unsterilized cheese whey, the column was washed thoroughly with deionized water. Next, the basic protein fraction adsorbed on the resin was eluted with a 0.02 M carbonate buffer (pH 7.0) containing 0.98 M sodium chloride. After adjusting the pH to 1.5 with hydrochloric acid, add pepsin (Kanto Chemical)
%, And the enzyme reaction was carried out with stirring at 37 ° C. for 1 hour.
After adjusting to 0.5% pancreatin (manufactured by Wako Pure Chemical Industries)
The mixture was added so as to have a concentration, and an enzyme reaction was carried out while stirring at 37 ° C. for 1 hour. The enzyme reaction was stopped by heating at 85 ° C. for 10 minutes to inactivate the enzyme. The hydrolyzate thus obtained is treated with an ultrafiltration membrane having a molecular weight fraction of 30,000, and the permeate is desalted and concentrated with a reverse osmosis membrane, followed by freeze-drying to obtain a powdery basic peptide fraction. 150g
I got

[0026]

Test Example 4 The basic peptide fraction obtained in Reference Example 2 was analyzed for its component composition. The protein was quantified by the Kjeldahl method, the fat was quantified by the Soxhlet extraction method, and the ash was quantified by the wet ash method. Table 4 shows the results.

[0027]

[Table 4] 水分 Water 1.0 (% by weight) Protein 98.0 Fat 0.2 Ash 0.1 Other 0.7 そ の 他──

[0028]

Test Example 5 The amino acid composition of the basic peptide fraction obtained in Reference Example 2 was analyzed. The amino acid composition was measured with an amino acid analyzer (Hitachi L-8500) after hydrolyzing the basic protein fraction with a 6N hydrochloric acid solution at 110 ° C. for 24 hours. Table 5 shows the results.

[0029]

[Table 5] ────────────────── Aspartic acid 10 (wt%) Serine 5 Glutamic acid 13 Proline 7 Alanine 5 Leucine 9 Lysine 10 Histidine 2 Arginine 9 Other 30 ─── ───────────────

[0030]

Example 1 80 kg skim milk whey protein concentrate (WP
C) 1.5 kg and lactose 15 kg were added and dissolved, and further, 25 g of casein dissolved with a predetermined amount of alkali was added and dissolved. The water-soluble vitamins component (vitamin B _1, vitamin B _2, niacin, vitamin B _6, biotin, folic acid, pantothenic acid, vitamin B _12, vitamin C, choline and inositol) and mineral components (calcium, phosphorus, chlorine, iron , Magnesium, potassium, copper, iodine,
(Zinc, manganese and selenium)
Further, 1 kg of the powdery basic protein fraction obtained in Reference Example 1 was added and dissolved. Next, fat-soluble vitamins
(Vitamin A, β-carotene, vitamin D, vitamin E
9 kg of prepared fat (mixed fat of palm oil, canola oil, soybean oil and fish oil) dissolved by adding and mixing, homogenized, sterilized, concentrated and dried for infants 30 kg of prepared milk powder was obtained.

[0031]

[Example 2] Whey protein concentrate (WP)
C) 0.7 kg and lactose 4.4 kg were added and dissolved, and further, 6 g of casein dissolved with a predetermined amount of alkali was added and dissolved. The water-soluble vitamins component (vitamin B _1, vitamin B _2, niacin, vitamin B _6, biotin, folic acid, pantothenic acid, vitamin B _12, vitamin C, choline and inositol) and mineral components (calcium, phosphorus, chlorine, iron , Magnesium, potassium, copper, iodine,
(Zinc, manganese and selenium) Add 0.06 kg and dissolve,
Further, 50 g of the basic peptide fraction obtained in Reference Example 2 was added and dissolved. Next, fat-soluble vitamins (vitamin A, β-carotene, vitamin D, vitamin E and vitamin K) were added and dissolved, and 2 kg of prepared fat (mixed fat of palm oil, canola oil, soybean oil and fish oil) was added. After mixing, homogenizing, sterilizing, concentrating and drying, 10 kg of infant formula was obtained.

[0032]

Test Example 6 A rat feeding test was performed using the basic peptide fraction obtained in Reference Example 2. Breastfeeding, WGWa
II method (Science, vol.109, p.1313, 1975).
Sixteen SD rats, 3 days old, were divided into two groups, and a polyethylene tube (SP10, inner diameter 0.28
mm, manufactured by Natsume Seisakusho Co., Ltd.) and adjusted to a 25% concentration with the formula powder shown in Table 6 by syringe pump.
(WPI) to feed the stomach. The amount of milk was adjusted according to the growth (1 ml / day to 10 ml / day). Then, feeding was performed until the age of 21 days, at which time the rat was dissected and the femur was removed, and a rheometer (RX-100, I-Techno) was used.
Was used to measure bone strength. The result is shown in FIG. According to this, it was recognized that the bone strength of the group of the present invention to which the basic peptide fraction was added was significantly higher than the bone strength of the control group.

[0033]

[Table 6] Composition of rat milk (g / 100g) ─────────────────────────────────── Control group Inventive group ─────────────────────────────────── Whey protein concentrate (WPC) 2.1 (g / 100g) 1.6 (g / 100g) Casein 34 34 Basic peptide fraction ─ 0.5 Lactose 10.36 10.36 Vitamin mixture ¹⁾ 0.08 0.08 Choline chloride 0.46 0.46 Mineral mixture ²⁾ 7 7 Oil mixture ³⁾ 46 46 ─────── ──────────────────────────── ¹⁾ Vitamin mixture contains vitamin A, vitamin D, vitamin E, vitamin K, vitamin C, vitamin B _1, vitamin B _2, vitamin B _6, vitamin B _12, niacin, pantothenic acid, mixtures of folic acid and biotin. ^{2) The} mineral mixture is a mixture of potassium carbonate, sodium chloride, calcium carbonate, sodium ferrous citrate, zinc sulfate, magnesium sulfate and copper sulfate. ^{3) The} fat mixture is a mixture of milk fat, palm oil, soybean oil, lard and lecithin.

[0034]

Test Example 7 An experiment was conducted in which the basic protein fraction obtained in Reference Example 1 was administered to rats in the late stage of suckling. Sixteen 15-day-old male SD rats were weaned early and divided into two groups.
Were administered until 28 days of age. In the case of a 15- to 28-day-old rat, in the case of a human, it corresponds to the period of drinking follow-up milk from the late suckling period to the early weaning period.

[0035]

[Table 7] 対 照 Control group Invention group ───────── ─────────────────────── Casein 20.0 (g / 100g) 19.0 (g / 100g) DL-methionine 0.3 0.3 Basic protein concentrate ─ 1.0 lard 19.0 19.0 Corn oil 1.0 1.0 Mineral mixture ¹⁾ 3.5 3.5 Vitamin mixture ²⁾ 1.0 1.0 Choline bitartrate 0.2 0.2 Cellulose 5.0 5.0 α-corn starch 15.0 15.0 Sucrose 35.0 35.0 ──────────────── ──────────────── ^{1), 2)} mineral mixtures and vitamin mixtures by the addition of AIN-76 ^TM.

At the age of 28 days, the rat was dissected, the femur was excised, and the bone matrix component was analyzed. The bone matrix component of the femur was hydrolyzed with a 6N hydrochloric acid solution at 110 ° C. for 24 hours, and then measured with an amino acid analyzer (Hitachi L-8500). Table 8 shows the results. According to this, it was confirmed that ingestion of the basic protein fraction from late suckling to early weaning increased the content of hydroxyproline and proline, which are components of the bone matrix, and promoted the growth of the bone matrix.

[0037]

[Table 8] 対 照 Control group Invention group 群ハ イ ド ロ Hydroxyproline 1,780 ± 45 (ppm) 1,890 ± 51 ^* (ppm) Proline 2,890 ± 60 3,010 ± 60 ^* Hydroxylysine 600 ± 21 610 ± 21 Lysine 2,300 ± 95 2,410 ± 95 ─────────────────────────────────数 値 Numerical values represent mean ± standard deviation. * Significantly different from control group (p <0.05)

[0038]

The infant formula containing the milk-derived basic protein and / or the milk-derived basic peptide of the present invention has the effect of promoting bone growth by oral administration, so that it can be used for artificial nutrition. Promote bone growth and prevent various bone diseases, particularly osteomalacia.

[Brief description of the drawings]

FIG. 1 shows a comparison of a bone strength of a rat to which a formula powder was administered in Test Example 6 with a control.

(56) References JP-A-7-95850 (JP, A) JP-A-5-202098 (JP, A) JP-A-2-303457 (JP, A) (58) Fields studied (Int .Cl. ⁷ , DB name) A23C 9/152-9/18 A23C 21/06 A23J 1/20 A23L 1/305 A61K 38/00

Claims (8)

(57) [Claims] 1. An infant formula having an osteogenesis-promoting effect by adding a milk-derived basic protein and / or a milk-derived basic peptide. 2. The infant formula according to claim 1, wherein the formula is a powdered milk. 3. The amount of the basic protein derived from milk and / or the basic peptide derived from milk is 100% per 100 g of the product.
3. The infant formula according to claim 2, wherein the amount is from 1 to 1,000 mg. 4. The infant formula according to claim 1, wherein the formula is a liquid formula. 5. The amount of the basic protein derived from milk and / or the basic peptide derived from milk is 13% per 100 ml of the product.
The infant formula according to claim 4, wherein the amount is from 130 to 130 mg. 6. The milk-derived basic protein is eluted with a salt concentration of 0.1 to 2.0 M after the milk or milk-derived raw material is brought into contact with a cation exchange resin to adsorb the basic protein. It is a basic protein fraction obtained,
5. The infant formula according to any one of 5. 7. A milk-derived basic peptide is contacted with milk or a milk-derived raw material on a cation exchange resin to adsorb a basic protein, and then eluted with an eluate having a salt concentration of 0.1 to 2.0 M. The infant formula according to any one of claims 1 to 5, which is a basic peptide fraction obtained by decomposing the obtained basic protein fraction with a protease. 8. The infant formula according to claim 7, wherein the milk-derived basic peptide is a basic peptide fraction having a molecular weight of 2,000 to 20,000 obtained using pepsin and pancreatin as proteolytic enzymes.