JPH06305956A - Protein absorbefacient and nutrient composition containing the same - Google Patents

Abstract

PURPOSE:To obtain a protein absorbefacient, containing a polyamine as an active ingredient and capable of enhancing the utilization efficiency of proteins and promoting the growth of the digestive tract. CONSTITUTION:The protein absorbefacient contains a polyamine which is a straight-chain aliphatic hydrocarbon having >=2 primary amino groups, especially spermine as an active ingredient. The polyamine comprising >=20% spermine is contained in an amount of 40mug at the minimum based on 1 g proteins in the composition, especially 8mug at the minimum in the case of the spermine based on 1g proteins in the composition. The absorption of the proteins can be enhanced to keep good growth and healthy conditions by blending the polyamine therein. The load on infants in metabolism can be reduced by preparing modified milk powder for the infants having the same protein content as that of mother's milk. Furthermore, the allergy can be prevented from being induced with an ingested food by the development of the digestive tract.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a protein absorption promoter containing polyamine as an active ingredient and a nutritional composition containing the same, particularly infant formula. Spermine is particularly effective as the polyamine in the present invention.

[0002]

2. Description of the Related Art The protein requirement of breast-fed infants is generally 2.5 g / kg body weight for infants one month old, and 1.5 g for the weaning period after six months of age. However, in the case of artificially fed infants, the type of protein that is ingested is different from that of breast milk, and the amino acid balance is inferior to that of breast milk, so the amount of protein required must be increased. As a result, high protein intake requires metabolism of amino acids that are wasted in excess of the required amount, which imposes a heavy burden on the infant and causes disorders such as fever, coma, diarrhea, edema, and metabolic acidosis. Can be seen. Furthermore, the urea level in blood may increase, the amount of the phenol derivative excreted in urine may increase, and the physiological metabolism of the infant may become abnormal. Therefore, attempts have been made to reduce the protein content of infant formula to the level of breast milk as much as possible. However, as a result, it has been reported that the weight gain of artificially fed infants was lower than that of breast-fed infants, and it is suggested that the protein content of infant formula powders currently on the market has reached the limit. According to the Japanese nutritional requirements reported by the Ministry of Health and Welfare, the protein requirement for artificially fed infants is 2
3.3 ± 0.1 g / kg body weight / day until 2.5 months, 2.5 in 2-5 months
± 0.1g / kg bw / day, 3.0 ± 0.1g / kg bw / day in 6 to 12 months. However, in Europe and America, it is lower than the Japanese standard, 2.2 g / kg body weight / day, 2.2 g / kg body weight / day, 2.0 g / kg body weight / day, respectively, and the protein content in infant formula ( 1.5g / 100ml or less) is less than domestic products. The reason for this is that, in Europe and the United States, the intake of breast-fed infants is used as the calculation standard for protein intake. However, the average protein concentration of breast milk (mature milk) is actually 1.35 g / 100 ml (minimum 1.1 to maximum 1.7 g / 100 ml), so the protein concentration of infant formula is about 1.7 g / 100 ml. It is considered that further lowering is necessary in designing an infant formula in which breast milk is ideal. Loennerdal et al. Have a protein concentration of 1.
It has been reported that there is no problem in weight gain of infants even when they are fed with 3 g / 100 ml of artificial milk (Acta Pediatr. Scand., 79,
257, 1990).

Thus, although breast milk has a lower protein content than infant formula, breast-fed infants show good growth. It is expected that this does not depend solely on the good quality of the protein, ie the amino acid composition being suitable for human growth.

On the other hand, recently, the prevalence of allergic diseases has increased, and the number of patients has increased, leading to social problems. Especially, food allergies that are common in infants and children
There has been much interest in combination with diseases such as atopic dermatitis and childhood asthma. The mechanism of food allergy development varies, but it is thought to occur mainly due to invasion of the antigenic allergen (antigen) from the undeveloped gastrointestinal mucosa of infants. In order to prevent or treat such food allergies, diet restrictions that prevent intake of allergen substances are most commonly performed. However, it has been clarified that these allergenic dietary ingredients include many high-quality proteins such as eggs and milk, and if such dietary restrictions are applied during the growing season, malnutrition prevents normal growth. Recently, it has been suggested that even with such an allergenic protein, an allergic reaction does not occur if the amount of intake is reduced and the protein is ingested successfully. Therefore,
It is expected that allergies can be prevented by early maturation of the digestive tract and by keeping protein intake low.

[0005]

DISCLOSURE OF THE INVENTION When the present inventors consider the amount of nitrogen and energy required by an infant in consideration of the digestive absorption rate of protein, there is a clear limit in calculation of the protein content of human milk, It was assumed that the non-protein nitrogen present in breast milk affected the digestion and absorption of proteins. And
Based on this assumption, the relationship between various non-protein nitrogen components in breast milk and the digestive absorption of proteins was examined, and it was found that polyamine in breast milk promotes the digestive absorption of proteins to form the present invention. Came to. Further, the inventors of the present invention have made earnest studies on the physiological effect of polyamine, and have found that spermine in polyamine is an active center component, and spermine alone can significantly enhance the effect as polyamine.

Polyamines are straight chain aliphatic hydrocarbons having two or more primary amino groups, such as putrescine, cadaverine, spermidine and spermine, which are usually contained in human milk at a concentration of 100 to 400 μg / 100 ml. There is. It is known that polyamines have the effect of promoting cell growth and differentiation, and in particular, polyamines taken orally have been reported to promote maturation of gastrointestinal mucosa (Grant, AL e.
t al., J. Anim. Sci., 68, 363-371, 1990; Dufour, C.
et al., Gastroenterology, 95, 112-116, 1988). However, when the polyamine was added to infant formula, etc., there was no report that the digestion and absorption of the protein contained therein was promoted, and the present inventors found it for the first time. Infant formula, an alternative to breast milk, has a low polyamine content, below 50 μg per 100 ml, and some products do not contain it at all. The reason for this is considered to be that the polyamine is also removed at the same time as desalting the whey as a raw material in the process for producing the modified milk powder. Therefore, by adding polyamine to infant formula and increasing the amount, not only nutritionally supplies a nitrogen source as a non-protein nitrogen component, but also by adding a polyamine's unique physiological effect, artificial nutrition The problem of protein content can be solved.

Of the above polyamines, spermine [N, N'-
Bis (3-aminopropyl) -1,4-diaminobutane] (N
H ₂ (CH ₂ ) ₃ NH (CH ₂ ) ₄ NH (CH ₂ ) ₃ NH ₂ ) has a primary amino group of 2
It is a straight-chain aliphatic hydrocarbon having 50 to 50
It is contained at a concentration of 200 μg / 100 ml. However, infant formula, a substitute for breast milk, has a low spermine content,
It is 8 μg or less per 100 ml, and some products do not contain it at all. Therefore, among polyamines, spermine, especially spermine, can be added to the infant formula to increase the amount of spermine's unique physiological effect and solve the problem of protein content in artificial nutrition.

[0008] That is, an object of the present invention is to provide a protein absorption promoter capable of enhancing the utilization efficiency of proteins and promoting the growth of the digestive tract. Furthermore, the subject of the present invention consists of a nutritional composition containing such a protein absorption enhancer, and when ingested, it is possible to enhance the absorption of protein and maintain good growth and health. Another object of the present invention is to provide a nutritional composition capable of preventing allergies caused by ingested food due to the development of the digestive tract.

[0009]

The present invention has been made to solve the above-mentioned problems, and the first invention relates to a protein absorption promoter containing polyamine as an active ingredient. In addition, the second invention relates to a nutritional composition containing polyamine, which improves the utilization efficiency of protein to maintain good growth and health and prevent food allergy. Further, the present invention relates to a protein absorption promoter and a nutritional composition containing spermine among polyamines as an active ingredient. As mentioned above, polyamines are expected to have a nutritional action as a non-protein nitrogen component to supply a nitrogen source necessary for infants. The feature of the present invention relates to a nutritional composition which has an effect different from the above-mentioned conventionally known effects, and in which the protein is effectively digested and absorbed even if the protein content is low, and good growth can be obtained.

The polyamine in the present invention is a general term for a straight-chain aliphatic hydrocarbon having two or more primary amino groups, and it is putrescine, cadaverine, spermidine, spermine, 1,3-diaminopropane, carzine and homo. Examples include spermidine, 3-aminopropylcadaverine, norspermine, thermospermine, and cardopentamine.
These are biogenic amines that are universally present in living organisms, but in the present invention, in particular, putrescine NH ₂ (CH ₂ ) ₄ NH ₂ , spermidine NH ₂ (CH ₂ ) ₄ NH (CH ₂ ) ₃ NH ₂ , spermine. NH ₂ (CH ₂ ) ₃ N
It is preferable to use H (CH ₂ ) ₄ NH (CH ₂ ) ₃ NH _2, and the use of spermine is particularly effective. As the polyamine in the present invention, it is possible to use one obtained by purifying polyamine and consisting only of spermine, or a mixture of spermine and other polyamines such as putrescine and spermidine.

These may be used as they are as chemical substances or in the state where salts such as phosphoric acid are bound, or they may be obtained from by-products when cheese is produced from milk such as milk. That is, the ultrafiltration permeate obtained when ultrafiltration of cheese whey as a by-product to obtain a whey protein concentrate (WPC) is brought into contact with a cation exchange resin to adsorb polyamine onto the cation exchange resin. Elution with an acid or salt solution, neutralization of the eluate and electrodialysis for desalting can be carried out to obtain a polyamine solution, which can be used as it is or after being subjected to a drying means such as spray drying. By doing so, not only the application of polyamine is expanded, but also milk can be highly utilized, which is industrially useful. Furthermore, it can be extracted from fish semen and the like by the same method.

As the protein absorbent in the present invention, polyamine may be used alone, but if necessary, vitamin, mineral, lactose and the like may be mixed and used. The dosage form is liquid, powdery, granular, or any other suitable state, and is added in an appropriate amount to foods, nutrients and the like.

The nutritional composition can be obtained by blending such a polyamine. The nutritional composition, as a milk substitute food, pathological food, baby food, nutritional drink, enteral nutritional agent, etc., especially added to the raw material during infant formula production,
It is desirable to obtain a polyamine-blended milk powder. As described above, the modified milk powder has a low polyamine content, and by adding polyamine, the digestive absorption of protein of the infant can be promoted and the allergy caused by food intake can be prevented.

That is, in the present invention, as shown in the following test examples and examples, by ingesting polyamine orally, the utilization efficiency of protein is enhanced, and good growth can be obtained even in a diet low in protein content. It was revealed that it will be done. The mechanism is that polyamine differentiates and matures the mucous membrane of the digestive tract to increase the peptidase activity present in the mucous membrane, and the dietary protein decomposed by pepsin and trypsin contained in the digestive juice is further decomposed into amino acids and oligosaccharides. It was confirmed by the present invention that the peptide is decomposed and is easily absorbed. As shown in Test Example 2, the fact that the addition of polyamine did not increase the weight simply because the nitrogen source increased was that spermine was added in an amount as small as 1 / 50,000 or less of the protein amount, as shown in Test Example 2. The weight has increased significantly, and this can be easily inferred. Further, it was revealed that in the case of a diet having a low protein content, at least 40 μg of spermine and polyamine containing 20% or more per 1 g of protein ingested, and particularly at least 8 μg of spermine, it was found to be effective. Even in products with the highest polyamine content in infant formulas currently on the market,
The amount of polyamine is 30 μg per 1 g of protein, of which spermine is only 4 μg, which is not sufficient to obtain the effect of the present invention.

Regarding the effect of preventing allergies, it was revealed that administration of polyamines, especially spermine, blocks the invasion of allergen into the body and suppresses the production of antibodies, which is one of allergic reactions. .
For this reason, it is expected that the mechanism by which the allergen does not enter the body due to the maturation of the digestive tract mucosa. As a raw material of the polyamine to be added, bovine,
Various milk-related materials and fish semen derived from milk of mammals such as goats and sheep can be used. In particular, the whey protein concentrate (WPC) is contained in a large amount in the residual fraction and the lactose fraction when the whey protein concentrate (WPC) is produced by an ultrafiltration membrane or an ion exchange resin. By using such a polyamine-rich raw material, it is possible to produce infant formula and the like containing polyamine. In particular, it is also possible to produce a infant formula containing 8 μg or more of spermine per 1 g of protein by using a raw material rich in spermine.

Next, test examples and examples which have led to the present invention will be shown.

[Test Example 1] Measurement of Polyamine Content of Breast Milk and Infant Formula Powder Polyamine content of breast milk and commercially available infant formula powder was determined by the method of Marton and Lee (Clin. Chem., 21, 1721-1).
724, 1975) and measured by a high performance liquid chromatography (HPLC) method using a Dionex HPIC-PAC ion exchange column. The measurement results are shown in Table 1.

[0017]

[Table 1] Polyamine content in milk ─────────────────────────────── Polyamine (μg / 100ml) ─────────────────────── Putrescine spermidine spermine ──────────────────────── ──────── Breast milk A 5 51 91 Breast milk B 100 132 168 ──────────────────────────────── Infant formula A 0 0 0 Infant formula B 27 15 8 ─────────────────────────────── Infant formula The polyamine concentration of milk was measured when the milk was prepared at a standard milk preparation concentration. Thus, it can be seen that the modified milk has a lower content of polyamines, especially spermine, than the milk.

[0018]

[Test Example 2] Low protein diet intake and weight gain: 5 Wistar rats (male, 4 weeks old), standard diet (milk casein 20%) group, low protein diet (milk casein 10%) group, low The animals were divided into protein diet + spermine (40 μg addition group, 80 μg addition group, 120 μg addition group) and bred for 28 days. When the body weight at the start of the experiment was compared with the body weight after 28 days, it was confirmed that the weight gain rate was remarkable in the group in which 80 μg or more of spermine was added to the low protein diet. The results are shown in Table 2.

[0019]

[Table 2] Breeding experiments ──────────────────────────────────── Sample intake Weight gain Weight gain / (g) (g) Protein intake (PER) ─────────────────────────────────── ── Standard diet group 402 ± 26 134.7 ± 6.4 1.68 ± 0.65 Low protein diet group 352 ± 12 107.4 ± 3.5 3.05 ± 0.20 Spermine 40 μg addition group 360 ± 15 110.5 ± 4.5 3.07 ± 0.11 80 μg addition group 376 ± 10 125.2 ± 3.0 3.33 ± 0.09 120 μg Addition group 380 ± 9 127.3 ± 2.9 3.35 ± 0.05 ─────────────────────────────────────

[0020]

[Test Example 3] Ingestion of low protein diet and increase in intestinal weight: The rat of Test Example 2 was bred overnight without feeding and water, and dissected the next day to remove the small intestine. After measuring the wet weight and length of the whole small intestine, it was dried in an oven set at 105 ° C. for 18 hours to measure the dry weight of the small intestine. As a result, the weight of small intestine per body weight was significantly high in the group to which spermine was added at 80 μg or more.
This means that the small intestinal mucosal tissue has grown significantly and matured. Table 3 shows the measurement results.

[0021]

[Table 3] Small intestine measurement results ──────────────────────────────────── Wet weight ( g / 100g body weight) Dry weight (g / 100g body weight) ──────────────────────────────────── Low Protein diet group 1.90 ± 0.12 0.42 ± 0.08 Spermine 40 μg addition group 2.11 ± 0.05 0.43 ± 0.04 80 μg addition group 2.42 ± 0.15 0.61 ± 0.10 120 μg addition group 2.39 ± 0.13 0.60 ± 0.13 ───────────── ───────────────────────

[0022]

[Test Example 4] Increase in intestinal mucosal peptidase activity: The small intestine of the rat of Test Example 2 was removed and the method of Kawakami and Loennerdal (Am. J. P.
hysiol., 261, G841, 1991) to prepare a small intestinal brush border membrane fraction. 7 by amide bond to carbosyl group of amino acid
Lys-Al, a peptide substrate (methylcoumarinamide: MCA) to which amino 4-methylcoumarin (AMC) is bound
Using a-MCA (manufactured by Peptide Institute, Inc.), the dipeptidyl aminopeptidase activity of the previously prepared small intestine brush border membrane fraction was measured. The AMC liberated by the enzymatic reaction was measured using a fluorometer (excitation wavelength: 380 nm; fluorescence wavelength: 440 nm). The measurement results are shown in Table 4. The measured value was shown as a relative value when the activity of the brush border membrane of the small intestine of the low protein diet group rat was set to 1. As a result, it was revealed that the enzyme activity of the group to which spermine was added at 80 μg or more was significantly high.

[0023]

[Table 4] Enzyme activity of brush border membrane of small intestine ──────────────────────────────── Dipeptidyl aminopeptidase activity ── ────────────────────────────── Low protein diet group 1 Spermine 40 μg addition group 1.2 ± 0.5 80 μg addition group 3.6 ± 0.9 120 μg addition Group 3.9 ± 1.2 ────────────────────────────────

[0024]

[Test Example 5] Allergen invasion inhibition effect: Wistar rats (14
(Age, 10) were divided into a control group and a spermine administration group. Both groups were weaned early and fed a powdered standard diet (20% milk casein). In the spermine administration group, 10 μl of the spermine solution (1 mg / ml) was orally administered every day on the 14th to 20th days using a micropipette. On the 21st day, 100 μl of β-lactoglobulin (βLg) solution (10 mg / ml) was orally administered,
Blood was collected after 1 hour and 2 weeks. On the other hand, anti-βLg serum was prepared by mixing βLg solution and Freund's complete adjuvant, emulsifying the mixture, and injecting it into three subcutaneous sites (both dorsal side and buttocks) of a 3-month-old rabbit (white Japanese breed, male, Kitayama Labes). Got Sandwich E with this antiserum as the primary antibody and a secondary antibody labeled with horseradish peroxidase (PO)
By the LISA method, the amount of βLg in blood was measured using blood after 1 hour. In addition, anti-βLgIgE in blood after 2 weeks
Was measured by ELISA using βLg and PO-labeled anti-rat IgE antibody (manufactured by Nordic). As a result, as shown in Table 5, it was found that the spermine-administered group had a significantly lower blood βLg and anti-βLgIgE content than the control group, and thus antibody production was suppressed and allergies could be suppressed.

[0025]

[Table 5] Blood βLg and anti-βLgIgE measurement results ─────────────────────────────── βLg (ng / ml) Anti-βLgIgE (ng / ml) ─────────────────────────────── Control group 30.6 ± 15.6 450.8 ± 108.6 Spermine administration group 5.7 ± 2.9 126.9 ± 86.5 ────────────────────────────────

[0026]

Example 1 (1) Preparation of Polyamine In the process of obtaining a whey protein concentrate (WPC) produced by concentrating undesalted cheese whey with an ultrafiltration (UF) membrane, a fraction that permeates the UF membrane I got 500 liters. This solution was passed through a column packed with a cation exchange resin Dowex 50-x8 (H ⁺ type) to adsorb polyamine. The resin was thoroughly washed with 0.7 M saline to remove impurities such as basic amino acids, and then polyamine was eluted with 6N hydrochloric acid. In the eluate
After 30% sodium hydroxide solution was added for neutralization, desalting was performed using an electrodialysis (ED) device to obtain a polyamine solution. This solution was spray dried to obtain 0.5 g of a polyamine raw material. This polyamine material contained 200 mg of spermine.
The above operation was carried out again to obtain 0.5 g of a polyamine raw material in the same manner. This material was used as a protein absorption promoter.

(2) Preparation of infant formula powdered milk containing polyamine 1 g of the polyamine material obtained in (1) above was dissolved in 700 kg of water together with 6.8 kg of casein, 70.6 kg of whey powder, and 1 kg of vitamin and mineral components. In addition, vegetable oil
After mixing 23.9 kg and homogenizing, 100 kg of milk powder was obtained through the steps of sterilization, concentration and drying. 100 g of the obtained milk powder had a protein content of 13.0 g and a polyamine content of 800 μg.

[0028]

Example 2 Preparation of Infant Formula with Spermine Formulation 0.4 g of spermine diphosphate (C ₁₀ H ₂₆ N ₄ 2H ₃ PO ₄ ; Sigma, S-4513) 0.4 g casein 6.0 kg whey powder 71.4 kg
, 700 kg water with 1 kg vitamins and minerals
Dissolved in. Further, 23.9 kg of vegetable oil was mixed and homogenized, and then sterilized, concentrated and dried to obtain 100 kg of milk powder.
The protein content in 100 g of the obtained milk powder was 12.5 g, and the spermine content was 130 μg.

[0029]

The effects of the present invention are as follows. 1) Good growth (weight gain) can be obtained even with a nutritional composition having a low protein content. 2) By making an infant formula having the same protein content as breast milk, the metabolic load of the infant can be reduced. 3) Preventing allergens from entering the body can prevent allergies. 4) Good growth (weight gain) can be obtained even if the content of protein serving as an allergen is reduced to prevent allergies.

Claims (7)

[Claims] 1. A protein absorption enhancer comprising a polyamine as an active ingredient. 2. The polyamine is spermine.
Accelerator as described. 3. A nutritional composition containing polyamine, which can maintain good growth and health by increasing protein utilization efficiency and prevent food allergy. 4. The polyamine is spermine.
The composition as described. 5. The composition according to claim 3, wherein the nutritional composition is infant formula. 6. The composition according to claim 3, wherein the polyamine containing 20% or more of spermine is contained in an amount of at least 40 μg per 1 g of the protein in the composition. 7. The composition according to claim 4, which contains at least 8 μg of spermine per 1 g of protein in the composition.