KR20070069123A - Calcium binding amino acid - Google Patents

Abstract

Amino acid-bound calcium prepared by simultaneously extracting peptide, calcium and phosphorous from animal bone tissue is provided, which has high safety to the human body and is useful as a treating agent for osteoporosis and a food additive for calcium reinforcement. First, 1 to 10% by weight of an edible organic acid and 1% by weight of a hydrolase are put into 1% by weight of a fish bone frame at the same time and reacted for 4 to 12hr to extract phosphorous and calcium. Then, the phosphorylated amino acid peptides are filtered and centrifuged to produce supernatant which is freeze dried or spray dried. Any one of shark cartilage, cow bone, pig bone, chicken bone, raw anchovy and dried anchovy is used instead of the fish bone frame. The edible organic acid is preferably acetic acid(5 to 10%, pH:2 to 3).

Description

Calcium Amino Acids {CALCIUM BINDING AMINO ACID}

Calcium is the most abundant mineral in the human body and has about 1200gr, which is about 2% of body weight in normal adults. 99% of the calcium in the body forms the skeleton and teeth, and only about 1% is responsible for the regulation of physiological activities such as muscle contraction and relaxation, regular heartbeat, blood coagulation, enzyme activation, intracellular stimulation and excitation delivery. Doing.

In particular, calcium plays an important role in reducing the incidence of chronic diseases such as osteoporosis, hypercholesterolemia, arteriosclerosis, hyperlipidemia, hypertension, and the like, and therefore, proper calcium nutrition should be maintained at all times. The daily recommended amount of calcium is about 700mg. It is very important to consume calcium through food, but in fact, even if you eat foods containing calcium, the absorption rate of calcium in the body is very low. Only 45% of the situation is ingested.

In order for calcium to be absorbed in the body, it must be present in soluble form when passing through mucosal cells in the small intestine where calcium is absorbed. However, because the small intestine is weakly alkaline, it is impossible for calcium, an essential mineral, to exist in an ionic state. Therefore, calcium cannot maintain the state of ions in the mucosa without relying on special structures. In other words, when water-soluble mineral ions produced in the stomach are not protected by a specially structured compound, they are combined with excess phosphoric acid during the duodenum, resulting in non-absorbing mineral calcium phosphate, which is released into the powder.

Representative commercial calcium agents are supplied in the form of insoluble calcium such as calcium carbonate, calcium phosphate, etc., which are chemical synthetic products, and water-soluble salts such as calcium chloride, calcium citrate, calcium acetate, and the like. It is widely supplied as an insoluble calcium agent in the form of simple processing by grinding and calcining.

Meanwhile, as calcium absorption accelerators used to improve the absorption rate of commercially available calcium agents in the human body, oligopeptides obtained by decomposing proteins with specific enzymes and phosphorylated proteins or peptides chemically modified with specific proteins or peptides are used. Attempts are being made. The most well-known technique is CPP (casein phospho peptide) made by hydrolyzing casein protein in milk with enzyme trypsin, which is phosphorylated with serine residue of casein, which is combined with soluble calcium in the small intestine to facilitate the absorption of the body. Naito et al .: Agricultural and Biological chemistry, 36: 409-, 1972.

In addition, trypsin decomposition of casein, a milk protein, has been shown to be effective in preventing the formation of insoluble substances such as calcium phosphate which is combined with metal ions such as calcium and iron, forms soluble complexes and is excreted outside the body (Reeves et al.). Science, 128: 474-. 1958).

As the above-mentioned prior art, for example, a method for producing fish bone powder obtained by separating bones from fish and drying them (Japanese Patent Laid-Open No. 2-231059, Japanese Patent Laid-Open No. 4-121166) is known. It takes a lot of energy to dry. In addition, after washing the fish bones in high pressure water to decompose the remaining protein by protein hydrolase, washing and removing the dried bones by vacuum heating evaporation method and then pulverized to produce fish bone meal (Japanese Patent Laid-Open 2-231059) is known, but the low calcium content of the product thus produced is a disadvantage. On the other hand, in order to solve the additional problem, Japanese Patent Laid-Open No. 6-319487 discloses a method for producing water-soluble calcium by softening by dipping fish bones in acetic acid, but the process is complicated and liquid beverage Since it cannot be used other than this, its use is limited and it is not widely used.

As a related prior patent technology in Korea, Patent No. 10-0403284 and Patent No. 10-0399722, etc. disclose a method of preparing fish bone meal, water-soluble calcium, and calcium absorption promoting peptides from fish bones, but specific hydrolase enzymes are known. The process of extracting tuna from internal organs is complicated, and in the case of water-soluble calcium, it is difficult to be widely used from an economic point of view because of low yield of commodities.

In addition, the prior art for producing a casein phospho peptide (CPP), a calcium absorption accelerator that is widely used around the world, there is a Japanese Patent Publication (Heb. 2-7616, 1990) and the like under US Patent No. 4,358,465 (1982), The technology differs only in the kind of enzymes used for the decomposition of sodium casein, separation and purification methods, but the principles and processes are the same. Therefore, the hydrolysis process, the separation and purification process, and the chemical synthesis of calcium are artificially added. It is combined with CPP and dried.

Technical challenge

Currently, natural calcium materials such as calcium carbonate (calcium carbonate), calcium calcium, shell calcium, eggshell calcium, etc. Calcium phosphate (Calcium phosphate) group includes cow bone powder, bone bone powder, but bone bone powder is a wave of mad cow disease, Eggshell calcium and shell calcium are rarely used as a warning about the heavy metal contamination and the problem of absorption rate. Insoluble calcium such as Calcium carbonate and Calcium phosphate, which are used as chemical compounds, precipitates when added to milk, exposing defects to product stability, whereas calcium lactate, calcium citrate, and calcium acetate (Calcium citrate) Water-soluble calcium, such as Calcium acetate, has a severe taste change when added over a certain amount, as well as fatal changes such as aggregation caused by reaction with casein protein in milk, viscosity increase during heat treatment, and bitter taste due to gelation. Has been exposed.

On the other hand, CPP (casein phospho peptide) is the only material widely used as a water-soluble calcium absorption accelerator, and there are two types of CPP basic type, which is an absorption promoting peptide, and high-grade type in which calcium is added by artificially adding calcium to the CPP basic type. However, due to the complicated and expensive manufacturing process, it has limitations that cannot be widely used.

Technical solution

The present invention is to solve the above-mentioned problems, the object of the present invention is a natural calcium material that has been proven safe, water-soluble, affects the stability of the product due to changes in taste, precipitation, bonding, etc. when added to the product To provide a calcium calcium amino acid that can be produced in a simple, efficient process.

Favorable effect

As described above, the amino acid calcium according to the present invention is safe from the fish bone frame, which is a natural calcium material, enables the production of calcium with amino acids combined with excellent absorption rate in the human body, and also has a high concentration that does not require the use of calcium absorption accelerators in combination. It is made of natural calcium.

1 is a graph showing the degree of hydrolysis according to conditions for determining the optimal extraction conditions of mineral calcium and amino acid peptide from the bone frame.

Figure 2 is a graph showing the recovery rate of calcium for each condition for the determination of optimal conditions for the production of amino acid calcium from the bone frame.

Figure 3 is a graph showing the results confirmed using I.R-spectrum phosphorylation of amino acid peptides to confirm the binding capacity of the amino acid peptide and the extracted calcium.

Best Mode for Carrying Out the Invention

In order to achieve the above object, the amino acid calcium according to the present invention is prepared by reacting an edible organic acid and a protein hydrolase at the same time in the bone frame, and phosphorus and calcium are eluted from the bone frame, and the protein is broken down into amino acid peptides. The amino acid peptide and the phosphorus bond is characterized in that the amino acid peptide is phosphorylated, and the phosphorylated amino acid peptide and calcium is produced by binding.

In addition, it is preferable to react by stirring 1% of the fish bone frame based on the 1-10% solution of the organic acid and 1% of the hydrolase based on the fish bone frame for 4 to 12 hours.

In addition, it is preferable that 1% of the fish bone frame and 1% of the hydrolase are stirred for 4 to 12 hours on the basis of the 1-10% solution of the organic acid.

In addition, the calcium-phosphorylated phosphorylated amino acid peptide is preferably filtered and centrifuged to take a supernatant and then lyophilized or spray dried.

In addition, it is preferable that any one of shark cartilage, right bone, pork bone, chicken bone instead of the bone frame.

Embodiment for Invention

Example 1 Preparation of Hydrolyzed Peptides and Extraction of Calcium from Fishbone Frame

Into a reactor equipped with a stirrer, 1% of a wet crushed hoki frame (based on dry weight) and 1% of a pepsin (based on a bone frame) based on 10% acetic acid solution (pH: 2.0-2.2) were added. . The reaction mixture was stirred at 40 ° C. for 12 hours with gentle stirring, and then filtered and centrifuged to obtain a supernatant to obtain a hydrolyzed peptide composition containing a large amount of minerals.

On the other hand, in order to obtain the optimum conditions for hydrolysis and mineral extraction, the control group without the enzyme and the control group without the organic acid (distilled water, pH adjustment) were set under the same reaction conditions, and then the hydrolysis and calcium recovery were compared.

As a result, as shown in FIG. 1, the hydrolysis rate of the fish bone frame was about 25% and 40% when the organic acid and the hydrolase were treated, respectively, but about 70% when the organic acid and the enzyme treatment were combined. (

On the other hand, the recovery rate of calcium, as shown in Figure 2, when comparing the recovery rate of calcium from the fish bone frame, when the combination of the organic acid and hydrolase, both the content and yield of calcium was high.

Example 2 Preparation of Phosphorylated Amino Acid Peptides (Calcium Binding Peptides: Amino Acid Peptides: C.B.P.P) from Calcium and Phosphorus Hydrolyzed Amino Acid Peptides

1% hoki frame (dry weight basis) and 1% pepsin (dry bone frame basis) wet-crushed in a reactor equipped with a stirrer based on 10% acetic acid solution (pH: 2.0-2.2) Was added. After gentle stirring, the reaction was carried out at 40 ° C. for 12 hours, followed by filtration and centrifugation. The supernatant was taken, and lyophilized to obtain a phosphorylated peptide composition.

On the other hand, in order to make a control of the non-phosphorylated peptide, a non-phosphorylated peptide composition was prepared simultaneously under the same reaction conditions using a substrate composed of muscle proteins (hoki, meat only, only) from which bone components were completely removed.

The calcium binding capacity of the hydrolyzed amino acid peptide was determined by the phosphorylation of the amino acid peptide and compared with the I.R-spectrum for the above compositions to confirm this.

As shown in FIG. 3, the difference between the non-phosphorylated and phosphorylated peptides was found at 1,000-1,300 cm −1 where the phosphate group was present, and the peaks were found in the phosphorylated peptide, although the peaks were not identified. That is, P = O bonds at 1,300cm-1, P-O-C bonds at 1,100-1,200cm-1, and -O-P bonded to alkyl groups showed absorption bands at 1,000cm-1. These bands were not found in non-phosphorylated peptides, but in all samples.

The present invention is applicable to shark cartilage, beef bone, pork bone, chicken bone, raw anchovy, dried anchovy, etc. other than the fish bone frame.

In addition, the present invention may be used to prepare amino acid calcium containing amino acid peptides derived from collagen which is a component of animal bone tissue and blood using animal bone tissue as well as animal bone tissue and blood.

In the present invention, fish bone frame (Fish Bone Frame; that is the muscle protein is coupled to the fish bone) is a raw material of natural calcium in the fish processing factory (after fish meat cut of fish cutlet products, tuna and salmon, etc.) Fish bones after the cutting process for sashimi) and the muscle proteins attached to them were used as the main targets and stored in a freezer at -30 ℃.

In the present invention, the extraction of calcium, phosphorus and organic amino acid peptides, which are inorganic components, is carried out by simultaneously combining the elution of calcium and phosphorus with an edible organic acid and the extraction process of the amino acid peptide with proteolytic enzymes. Bone in vivo is composed of collagen and non-collagen protein and hydroxyapatite (Ca10 (PO4) 6 (OH) 2), which is an inorganic binder, and collagen constituting bone is a strong enzyme with hydroxyapatite, a constituent mineral, Although it is hard to be decomposed by hydroxyapatite, calcium and phosphorus, which are the basic constituents of hydroxyapatite, are not only eluted by organic acids, but also collagen is easily extracted by acetic acid, and muscle and bone proteins are effectively decomposed. It can be easily recovered along with the proteolytic products.

The edible organic acid used in the present invention is most preferably, but not limited to acetic acid (5-10%, pH: 2-3), edible lactic acid, citric acid, formic acid and the like.

As hydrolase used in hydrolysis with organic acids, pepsin or trypsin is used, and all enzymes that do not lose their activity in an acidic region satisfying the above conditions are applicable thereto.

Substrate concentration is 1-10% of organic acid solution, enzyme concentration is 1-5%, and hydrolysis and extraction time is 4-12 hours. The conditions, etc., shall be determined within the scope in consideration of economic feasibility.

Calcium and phosphorus eluted in the above extraction process are eluted and directly involved in the phosphorylation of amino acid peptides, which are partial hydrolysates of proteins produced by enzymatic decomposition, to form phosphorylated amino acid peptides, and to form phosphorylated amino acid peptides. The strong binding force absorbs the eluted calcium, producing the amino acid calcium.

Separation and purification of Calcium Binding Phospho Peptide (CBPP) formed through the extraction and binding process is performed by filtration system, and only lyophilization or spray drying by collecting only calcium binding phosphate peptides with a molecular weight of 10,000 Da or less. Prepared.

CBPP prepared according to the method of the present invention is a highly concentrated natural water-soluble calcium agent in the form of a combination of minerals such as calcium and amino acids, and is a very excellent material in terms of safety, and does not require the use of a calcium absorption accelerator as a food or beverage. It can also be used as a raw material for dietary supplements and pharmaceuticals.

Claims (5)

When edible organic acid and protein hydrolase are added to the bone frame at the same time, phosphorus and calcium are eluted from the bone frame, and when the protein is decomposed into amino acid peptides, the amino acid peptide and the phosphorus are combined to phosphorylate the amino acid peptide. Amino acid calcium, which is produced by combining phosphorylated amino acid peptide and calcium. The method of claim 1, Calcium amino acid, characterized in that 1% of the fish bone frame based on the 1-10% solution of the organic acid, 1% of the hydrolase on the basis of the fraudulent bone frame was stirred for 4 to 12 hours The method according to claim 1 and 2, The calcium-phosphorylated amino acid peptides were filtered and centrifuged to obtain supernatant, followed by lyophilization or spray drying. The method of claim 1, Amino acid calcium, characterized in that any one of shark cartilage, beef bone, pork bone, chicken bone, raw anchovy, dried anchovy instead of the fish bone frame. The method of claim 1, An amino acid calcium comprising an amino acid peptide derived from collagen which is a component of the animal bone tissue and blood using animal bone tissue and blood instead of the bone frame.

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