JP4451368B2 - Mineral absorption enhancer containing phosphorylated lactose, method for producing the same, and mineral-enriched food composition containing the same - Google Patents

Description

  The present invention relates to a mineral absorption enhancer containing phosphorylated lactose, a method for producing the same, and a food composition in which a mineral component containing the same is fortified.

  In recent years, the number of lifestyle-related illnesses from middle to old age has increased even among young people. It is said that the factors overlap. In particular, due to recent changes in eating habits, deficiencies occur due to lack of minerals in the diet, causing various diseases in a wide range of ages.

  Minerals are essential nutrients for living, but they must be taken from food because they cannot be made in the human body. However, in modern life, it has become difficult to compensate for the lack of minerals only from food. The reasons for this are (1) the use of chemical fertilizers and agricultural chemicals, the effects of air pollution and acid rain, etc., causing the mineral balance in the soil to be lost, and the resulting decrease in the mineral content in crops. For example, the unbalanced diet of modern people has been widespread by frozen processed foods, fast foods, and soft drinks. In addition, the fact that minerals are inherently difficult to be absorbed by the body is another factor that causes mineral shortages.

The diseases caused by these mineral deficiencies include the following.
Calcium (Ca): Osteoporosis, insomnia, muscle spasms, heart motives.
Iron (Fe): Iron deficiency anemia, chronic fatigue, dyspnea, constipation, stiff shoulders.
Magnesium (Mg): heart disease, psychosis, muscle pain, emotional instability, hypotension.
Zinc (Zn): high cholesterol, fatigue, loss of appetite, taste disorder, poor blood circulation, delayed recovery of injury, atopic dermatitis.
Chromium (Cr): Diabetes, hypertension, arteriosclerosis.
Selenium (Se): cardiac dysfunction, menopause, reduced anti-cancer power, cataracts, pancreatic function decline, infection, reduced male fertility.
Copper (Cu): Osteoporosis, anemia, respiratory abnormalities, depression, diarrhea.
Manganese (Mg): Diabetes, dizziness, hearing loss, ataxia.

  Among these, iron deficiency is known as the three major nutrient deficiencies, and is said to affect about 2 billion people worldwide, and is particularly common among infants, infants and women of fertile age. In particular, the iron deficiency in women is serious, and about 10% of adult women are said to have iron deficiency anemia, and about 40% have iron deficiency without any subjective symptoms (anemia reserve). Iron deficiency can cause anemia, which can cause childbirth of low-weight infants in pregnant women with severe iron deficiency and delayed development in children.

  With respect to diseases that are such social problems, interest in prevention is increasing from a viewpoint different from treatment of diseases, and improvement of dietary habits is attracting attention as primary prevention of diseases. Food has a primary function to maintain life (nutrition), a secondary function to enjoy meals (taste), and tertiary functions to maintain health such as rhythm adjustment of physical condition, biological defense, disease prevention, disease recovery, and aging prevention There is (biological regulation). There is “health functional food” as a food focusing on this tertiary function, and this health functional food is newly established alongside “special health food” under the health functional food system newly introduced in April 2001. It was. If certain conditions are met, it can be advertised and promoted as a nutritional functional food, which contains 12 cheek vitamins and 5 minerals (calcium, iron, zinc, copper, magnesium).

  In recent years, the development of health functional foods (nutrient functional foods) for the purpose of preventing and improving mineral deficiency has been actively conducted and attracted attention. In order to prevent diseases caused by mineral deficiencies, it is convenient and important to take minerals from foods without difficulty on a daily basis. Various compounds have been investigated for this purpose. For example, a technique is known that does not insolubilize minerals such as calcium and iron using starch-derived phosphorylated saccharide having at least one phosphate group in the molecule (for example, Patent Documents 1, 2, and 3). However, these phosphorylated saccharides require a complicated process for production, and use an enzyme reaction, so that there is a problem that it is difficult to control the reaction.

  In addition, a casein phosphopeptide (CPP) capable of forming a compound with calcium (Patent Document 4, Non-Patent Document 1, Non-Patent Document 2), a calcium citrate / calcium malate complex (Patent Document 5), etc. are also developed. However, these compounds are not always satisfactory in terms of price, flavor and mineral binding.

  The above-mentioned techniques are mainly examined mainly for ingesting calcium among minerals. Especially for iron in minerals, heme iron, iron pyrophosphate, iron lactate, ferrous citrate, EDTA Functional foods with enhanced iron content in the form of iron and the like have been developed. However, these compounds have been hindered by low palatability, which deteriorates the flavor and aroma of foods added, particularly beverages.

  Moreover, about the use of sugar phosphate ester, the pH buffer method using the same (patent document 6), the flavor improvement agent for heat cooking food (patent document 7), the additive for the flavor and texture improvement of retort cooked rice (Patent Document 8), mineral taste improvers (Patent Document 9) that improve the taste and texture of mineral components contained in foods are known, but all retain minerals such as iron. It is not intended to be introduced into food, and there is no disclosure or suggestion of the effects of the present invention.

JP-A-8-104696 JP 2002-20398 A JP 2002-37796 A JP-A-3-240470 JP-A-56-97248 JP-A-62-243675 Japanese Patent Laid-Open No. 5-95765 JP-A-8-1331099 JP 2003-79337 A Naito Hiroshi; Journal of Japanese Society of Nutrition and Food Science, 39, 433-439; 1986 Li Lian, Park Soga, Hiroshi Naito; Journal of Japanese Society of Nutrition and Food Science, 45, 333-338; 1992

  Therefore, in the present invention, in order to solve the deficiency due to lack of minerals, a compound having a so-called mineral carrier effect ("mineral absorption enhancer") that can be added to food so that minerals can be easily ingested and does not impair the flavor of the food. It is an object of the present invention to provide.

  Therefore, as a result of intensive studies to solve the above problems, the present researchers have made phosphorylated lactose excellent by chemically phosphorylating lactose, which is a natural component contained in milk and available at low cost. It discovered that it had mineral carrier ability, and came to complete this invention.

  That is, this invention can solve the said subject by providing the foodstuff composition which the mineral absorption enhancer containing phosphorylated lactose, its manufacturing method, and the mineral component containing it strengthened.

  The present invention provides a mineral absorption enhancer comprising phosphorylated lactose.

  The phosphorylated lactose contained in the mineral absorption enhancer according to the present invention preferably has at least 1 to 3 phosphate groups.

  The mineral to which phosphorylated lactose binds is preferably at least one mineral selected from calcium, iron, zinc, copper, and magnesium, and most preferably iron.

  The present invention also includes a step of dissolving or suspending a predetermined amount of lactose in a phosphate buffer, a step of drying the obtained solution or suspension, and a dried product at 70 to 150 ° C. at normal pressure. And a method of producing a mineral absorption enhancer comprising phosphorylated lactose, comprising the step of heating to a temperature of a predetermined time.

  In this invention, it is preferable that the heating temperature of the said heating process is 90 to 130 degreeC.

  In the production method according to the present invention, the phosphorylated lactose preferably has at least 1 to 3 phosphate groups.

  In the production method according to the present invention, the mineral to which the phosphorylated lactose binds is preferably at least one mineral selected from calcium, iron, zinc, copper, and magnesium, and most preferably iron. .

  Moreover, this invention provides the mineral enriched food composition containing the said mineral absorption enhancer.

  Since the phosphorylated lactose in the present invention is very excellent in mineral carrier ability as a mineral absorption enhancer, it can stably hold minerals required for living bodies and include them in foods.

  Lactose, which is a raw material for phosphorylated lactose in the present invention, is a natural component contained in milk, and can be obtained at a low cost, and phosphorylated lactose is synthesized by a chemical method using a safe reagent. It is easy to manufacture and it is easy to control the manufacturing process, which is economically and industrially advantageous.

  Moreover, since the mineral absorption enhancer which consists of phosphorylated lactose of this invention does not impair the flavor of the foodstuff used as object, it does not impair the palatability of foodstuff itself. Therefore, when added in the state of a conventional compound such as iron, generally, a mineral component that impairs the flavor of the food can be included in the food in an easy-to-drink state.

  Hereinafter, the present invention will be described with specific examples, but the present invention is not limited thereto.

  The raw material lactose used in the present invention is not limited to any kind as long as it is a quality sugar that can be used as a food, regardless of whether it is natural or synthetic.

  The phosphorylated lactose which is the mineral absorption enhancer of this invention can be obtained by introduce | transducing a phosphoric acid group into the lactose used as a raw material (phosphorylation). At this time, phosphorylation includes a chemical synthesis method, a biochemical synthesis method using an enzyme, and the like. In the present invention, phosphorylation is preferable.

Particularly, in the present invention, a method obtained by partially improving the method of Edward et al. (Edward Tarelli and Susan F. Wheeler; Analytical Biochemistry, 222, 196-201; 1994) is preferably used.
For example, a predetermined amount of lactose is dissolved or suspended in a phosphate buffer. Next, the obtained solution or suspension is dried. Phosphorylated lactose can be obtained by heating this to a predetermined temperature at normal pressure.

  The phosphate used in the phosphate buffer used in the production method of the present invention may be any of orthophosphate, pyrophosphate, and metaphosphate. The adjustment of the phosphate buffer, the method of dissolving (suspending) lactose in the phosphate buffer, etc. are not particularly limited, and can be performed according to a conventional method.

  The method for drying the lactose phosphate buffer solution or suspension is not particularly limited, and examples thereof include natural drying, hot air drying, and freeze drying. Among these, freeze-drying is particularly preferable.

As heating conditions under normal pressure after drying, the heating temperature is 70 ° C. to 150 ° C., preferably 80 ° C. to 140 ° C., more preferably 90 ° C. to 130 ° C. The heating time is 1 to 100 hours, preferably 24 to 72 hours.
At this time, the pH of the system is preferably 7 or less, and more preferably 2 to 5.5.

  Although there is no restriction | limiting in particular in the number of the phosphoric acid groups of the phosphorylated lactose in this invention, It is preferable that 1-3 residues are introduce | transduced. The number of these phosphate groups can be specified by a known method.

  In this invention, a mineral is hold | maintained with phosphorylated lactose, and it is possible to make a foodstuff contain a mineral, without impairing flavors, such as a foodstuff. Examples of minerals include calcium, magnesium, iron, copper, and zinc, with iron being particularly preferred. In the case of retaining the mineral, it is not limited to this, but it can be combined with phosphorylated lactose as an inorganic ion during the synthesis of phosphorylated lactose or subsequently after the synthesis is completed.

  The form of these minerals is not particularly limited. For example, as a donor inorganic salt that can be dissolved in a solution having a physiological pH and can be dissociated in the solution to give ions of each mineral (for example, monovalent or divalent ions in the case of iron). It can be used in the production of the mineral absorption enhancer of the present invention. In particular, the iron-saturated mineral absorption enhancer according to the present invention can be obtained by freeze-drying a compound in a very dilute iron ion solution.

  It is possible to use the mineral absorption enhancer which consists of phosphorylated lactose of this invention directly for the composition for food or drink, or for the composition for food additives. Although it does not specifically limit as a food-drink composition, In addition to general food as food, special-purpose foods (food for the sick, health functional foods (specific health foods and nutritional functional foods)), supplements, treatment Food can be given. Examples of drinks include milk drinks and soft drinks. The present active ingredient can be used as it is, or can be used according to a conventional method for ordinary food compositions such as mixing with other foods or food ingredients. In addition, the state of the food or drink normally used, for example, any of solid (powder, granule, etc.), paste, liquid or suspension may be used. Addition and processing methods to foods are known.

  The mineral absorption enhancer containing phosphorylated lactose according to the present invention can be used by adding to a food composition. The amount of the mineral absorption enhancer added to the food composition can be arbitrarily determined according to its purpose and use. Although this invention is not limited to this, As the content, 0.001 to 100% (w / w) normally with respect to the whole quantity, Especially 0.1 to 100% is preferable.

  Other components that can be used together with the mineral absorption enhancer containing phosphorylated lactose according to the present invention are not particularly limited, but include water, proteins, carbohydrates, lipids, vitamins, organic acids, organic bases, fruit juices, flavors, and the like. Can be used as the main component. Examples of the protein include whole milk powder, skim milk powder, partially skim milk powder, casein, whey powder, whey protein, whey protein concentrate, whey protein isolate, α_casein, β-casein, κ-casein, β-lactoglobulin, α-lactalbumin, lactoferrin, soy protein, egg protein, meat protein and other animal and vegetable proteins, hydrolysates thereof; butter, milk minerals, cream, whey, non-protein nitrogen, sialic acid, phospholipid, lactose, etc. Examples include various milk-derived components. Examples include saccharides, processed starch (in addition to text phosphorus, soluble starch, British starch, oxidized starch, starch ester, starch ether, etc.), dietary fiber, and the like. Examples of the lipid include animal oils such as lard, fish oil, etc., fractionated oils, hydrogenated oil, transesterified oil, etc .; palm oil, safflower oil, corn oil, rapeseed oil, coconut oil, fractionated oils thereof, Examples include vegetable oils such as hydrogenated oils and transesterified oils. Examples of vitamins include vitamin A, carotene, vitamin B group, vitamin C, vitamin D group, vitamin E, vitamin K group, vitamin P, vitamin Q, niacin, nicotinic acid, pantothenic acid, biotin, inositol, choline. And folic acid. Examples of the organic acid include malic acid, citric acid, lactic acid, and tartaric acid. These components can be used in combination of two or more, and synthetic products and / or foods containing a large amount thereof may be used.

  Hereinafter, the present invention will be described in detail with specific examples, but the present invention is not limited thereto. In the experiments, special grade or general reagents from Wako Pure Chemical Industries, Ltd. (Osaka) were used unless otherwise specified.

(Phosphorylation of lactose)
Phosphorylation of lactose was performed by the phosphate buffer method, which is a partially improved method of Edward et al. That is, 200 mg of lactose was suspended in 10 ml of 0.1M KH ₂ PO ₄ —Na ₂ HPO ₄ buffer (pH 5.5) and heated in a 70 ° C. water bath for 30 minutes. Thereafter, it was lyophilized in a threaded test tube. This was then opened and heated with a heating block under normal pressure. The phosphorylation reaction was heated at 80, 100, or 120 ° C. for 24, 48, or 72 hours.

(Gel filtration)
In order to desalinate the obtained heated sample, gel filtration chromatography with TOYOPEARL HW-40S was performed. Below, the conditions of gel filtration are shown. The sample concentration was 200 mg / 3 ml of the heated sample.
Column: TOYOPEARL HW-40S (2.6 × 90 cm) (Tosoh Corporation)
Mobile phase: Milli-Q water flow rate: 0.7 ml / min
Fractionation: 10 ml / main detection: phenol sulfate method (490 nm, neutral sugar)
It fractionated into peak A (Fraction No. 30-40) and peak B (Fraction No. 50-60). The chromatogram at each temperature and reaction time is shown in FIG. 1 (Reaction conditions A: 80 ° C., 72 hours, B: 80 ° C., 7 days, C: 100 ° C., 72 hours, D: 100 ° C., 5 days, E: 120 ° C. 24 hours). In the figure, the area of peak A was the largest at 120 ° C. and 24 h when compared. What recovered this peak A was designated as crude phosphorylated lactose (hereinafter abbreviated as P-Lac as appropriate).

(Anion exchange chromatography)
The desalted crude P-Lac was subjected to anion exchange chromatography using DEAE-TOYOPEARL 650M for measurement. The conditions for anion exchange chromatography are shown below.
Column: DEAE / TOYOPEARL 650M (1.6 × 15 cm) (Tosoh Corporation)
Mobile phase: 50 mM Tris.HCl buffer (pH 8.6)
Elution: Linear gradient elution flow rate with 0-1M NaCl solution in the same buffer: 1 ml / min
Fractionation: 10 ml / main detection: phenol sulfate method (490 nm, neutral sugar)
FIG. 2 shows the results obtained by separately collecting peak A and peak B in gel filtration and subjecting them to anion exchange chromatography. In peak A, one peak was observed in the non-adsorbed fraction and one peak in the adsorbed fraction. However, in peak B, only one peak was observed in the non-adsorbed fraction. The peak of the non-adsorbed fraction was found to be a lactose peak having no charge, and the peak of the adsorbed fraction was a negative P-Lac peak. From this, it was found that peak A contained P-Lac.

(Quantitative determination of phosphorus)
Samples were taken into test tubes and lyophilized. To this, 0.4 ml of perchloric acid (70%) was added and heated using an electric stove in an open system until the sample became transparent (about 20 minutes). After completion of the decomposition, 2.4 ml of ammonium molybdate reagent [4.4 g of special grade ammonium molybdate dissolved in 200 to 300 ml of distilled water, and 14 ml of special grade sulfuric acid was added thereto to make 1 L] and 2.4 ml of reduction [Fiske & SubaRow Reducing reagent: 30 g of sodium hydrogen sulfite, 6 g of anhydrous sodium sulfite, and 1-amino-2-naphthol-4-sulfonic acid (special grade) O.D. 5 g was ground and mixed in a mortar, made 250 ml, allowed to stand in the dark for 3 hours, and then filtered into a brown bottle. This was diluted 1:12 and used. And then heated at 100 ° C. for 10 minutes. After cooling, the absorbance at 830 nm was measured. The phosphorus content contained in the sample was calculated by a calibration curve prepared using potassium dihydrogen phosphate.
It was 13.2% when the phosphorus content of P-Lac obtained from 120 degreeC and 24 h heating conditions was measured. When one residue of a phosphate group was introduced into lactose, the phosphorus content was 7.4%, 12.4% when two residues were introduced, and 16.1% when three residues were introduced. From this, it was found that in P-Lac heated at 120 ° C. for 24 h, two phosphate groups were introduced into lactose on average.

(Mass analysis by fast atom collision mass spectrometry (FAB-MS))
MilliQ water was selected as the measurement solvent, and glycerol was selected as the measurement matrix. High-performance double-convergence mass spectrometer JMS / 700 type (JEOL Ltd.) based on Fast Atom Bombardment-Mass Spectrometry (hereinafter referred to as FAB / MS) using POSITIVE mode and various conditions with low resolution. Was used for mass spectrometry.
The mass spectrum of the obtained P-Lac is shown in FIG. Since the measurement is performed in the POSITIVE mode, the molecular weight is obtained by adding the proton component (H +, mass 1) to the actual molecular weight. The molecular weight of lactose is 342, and the molecular weight increases by 79 when one residue of a phosphate group is introduced. The peak at 445 is obtained by introducing one phosphate group into lactose, the peak at 525 is obtained by introducing two phosphate groups into lactose, and the peak at 625 is obtained by introducing three residues. Represents. In FIG. 3, since peaks were observed at 445, 525, and 625, it was found that the number of phosphate groups introduced into lactose was mainly composed of introduced molecules of 1 to 3 residues.

(Measurement of mineral carrier ability of phosphorylated lactose using BIACORE)
Materials: Sensor chip SA (streptavidin is introduced on the surface and a biotinylated ligand is immobilized by a biotin-avidin bond), HBS-EP buffer (both Pharmacia Biotech (Sweden)), Biotin- PV-LA (Seikagaku Corporation)
(1) Phosphorylation of PV-LA-Biotil Biotin-PV-LA was phosphorylated by the above method.
(2) Immobilization of ligand (PV-Suger-Biotin) on sensor chip SA Biotin-PV-LA and Biotin-PV-phosphorylated LA to be used were prepared with 0.1 mg / ml HBS-EP buffer. This solution was added to the sensor chip SA and immobilized by biotin-avidin reaction.
(3) Preparation of analyte (metal ion) Ferrous sulfate (FeSO ₄ · 7H ₂ O, hereinafter referred to as iron ion) is prepared as a metal ion in 0.3 mg / ml HBS-EP buffer and solubilized. After confirming that, it was set as the analyte.
(4) Mineral ion binding test for PV-Suger-Biotin BIACORE measurement conditions Running buffer: HBS-EP buffer sample addition amount: 10 μ1
Flow rate: 3μ1 / min
Temperature: 25 ° C
FIG. 4A shows the result of the confirmation test of the amount of ligand binding to the sensor chip SA for Biotin-PV-phosphorylated LA at 120 ° C. and 24 hours heating, and B) shows the biotin-PV-LA. FIG. 5 shows the result of flowing ferrous sulfate as the analyte with the ligand bound to the sensor chip SA. In FIG. 5, A) shows the iron ion binding property to Biotin-PV-phosphorylated lactose, and B) shows the iron ion binding property to Biotin-PV-lactose. Table 1 summarizes the results.

  In Biotin-PV-phosphorylated LA phosphorylated on heating at 120 ° C. for 24 hours, iron ions showed high binding with 1400 RU, and it was confirmed that phosphorylated lactose according to the present invention has a high mineral carrier ability. .

  The phosphorylated lactose Fe obtained above was purified and added to soft drinks and food compositions. The amount added was 1%. When the sensory test was done about the fragrance and flavor with respect to the obtained soft drink and food composition, 19 out of 20 people evaluated it as equivalent to the comparative non-added product. Therefore, it was found that the mineral absorption enhancer comprising the phosphorylated lactose of the present invention can contain minerals without impairing its flavor even when added to food.

  ADVANTAGE OF THE INVENTION According to this invention, the mineral absorption enhancer containing phosphorylated lactose which has high mineral carrier ability can be provided from easily available lactose as a raw material. By using the said mineral absorption enhancer, it becomes possible to strengthen the mineral component of the food / beverage product composition without impairing the flavor of the food / beverage product composition to be added. In addition, the mineral absorption enhancer can be produced simply by a safe chemical method and in an economically and industrially advantageous manner.

The figure which shows the result of the isolation | separation by the gel filtration chromatography of phosphorylated lactose (Reaction condition A: 80 degreeC, 72 hours, B: 80 degreeC, 7 days, C: 100 degreeC, 72 hours, D: 100 degreeC, 5 days, E: 120 ° C., 24 hours). The figure which shows the isolation | separation result by the anion exchange chromatography of phosphorylated lactose. Mass spectrum of phosphorylated lactose by FAB-MS. A) is a graph showing the binding property of Biotin-PV-phosphoractose to the sensor chip SA, and B) is a graph showing the binding property of Biotin-PV-lactose (control) to the sensor chip SA. A) is a graph showing the iron ion binding property to Biotin-PV-phosphorylated lactose, and B) is a graph showing the iron ion binding property to Biotin-PV-lactose.

Claims (3)

A step of dissolving or suspending a predetermined amount of lactose in a phosphate buffer, a step of drying the resulting solution or suspension, and heating the dried product to a temperature of 70 ° C. to 150 ° C. at normal pressure for a predetermined time And a method for producing iron-bonded phosphorylated lactose comprising a step of mixing with iron ions . The manufacturing method of the phosphorylated lactose of Claim 1 whose heating temperature of the said heating process is 90 to 130 degreeC. The method for producing phosphorylated lactose according to claim 1 or 2, wherein the phosphorylated lactose has at least 1 to 3 phosphate groups.

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