KR100399722B1 - Preparation of fish bone powder and calcium absorption accelerating peptide using the enzyme from fish processing waste - Google Patents

Abstract

In the present invention, after a fish fillet operation in a fishery processing plant, a fish residue produced as a by-product is reacted with an enzyme extracted from fish intestines (tuna, mackerel, etc.), and then a large amount of calcium is contained in the reaction precipitate. The present invention relates to a method for producing fish bone meal and producing a peptide component capable of promoting calcium absorption from the reaction supernatant.

Fish bone meal and calcium absorption promoting peptides prepared according to the present invention can be used as a raw material for food, beverages, pharmaceuticals and the like for the supply of calcium and calcium absorption promotion.

Description

Preparation of fish bone powder and calcium absorption accelerating peptide using the enzyme from fish processing waste}

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to fish bone meal and calcium absorption promoting peptides prepared from residues of fish produced as by-products after filleting of fish in aquatic product processing plant. After reacting the proteolytic coenzyme, the present invention relates to a method for producing a fish bone meal containing a large amount of calcium component from the reaction precipitate, and a peptide capable of promoting absorption of calcium from the reaction supernatant.

In terms of the use of aquatic products in Korea, the total production (1998) is 2.83 million tons, and the total supply of imports and the previous year's supply is 407 million tons. In addition, the processing rate of the fish processing plant is 87%, and a large amount of fish processing residues such as intestines, scales, fish bones, and fish are generated. Currently, only a part of these fish processing residues are used as feed. It is disposed of, causing environmental pollution.

Therefore, the research which can utilize these aquatic processing residue is active trend. For example, crustacean shell is used as an extract material for chitin and chitosan, which are used in various industries, medicines and foods. From extraction of gelatin from fish (Kang et al., 1992, Bull.Kor.K. Fish. Soc., 25, 93-102; Kim et al., 1993, J. Korean Agric.Chem. Soc., 36 (6), 440-448; Kim et al., 1995, J. Kor. Ind. Eng. Chemistry, 6, 984-996; Kim et al., 1996, J. Kor. Fish.Soc., 29, 246-255), and from the bones the production and use of hydroxyapatite (Lee et al., 1997, J. Kor. Fish. Soc., 30, 652-659; Kim et al., 1997, J Kor. Ind. Eng. Chemistry, 8, 1000-1005; Kim et al., 1998, J. Kor. Ind. Eng. Chemistry, 9, 322-329). Studies on proteolytic enzymes for industrial applications from fish gut (Simpson et al., 1990, J. Food Sci., 55, 959-961; Haard and Simpson, 1994, Biotechnological Applications, AM Martin ed., Chapman Hall , London, pp. 132-154; Kim et al., 1997, Fish. Sci. 63, 421-427), as well as enzymes with a number of unique abilities (Ragnar et al., 1991, Biochem. Physiol., 99B, 755 ~ 761; Myrnes and Johansen, 1994, Preparative Biochem., 24, 69 ~ 80).

The present invention is also the result of research on the use of such aquatic processing residues.

In the human body, calcium accounts for about 1.5 to 2.0% of the body weight and is an essential nutrient for body formation and maintaining health, and lack of calcium inhibits bone growth, making it difficult to form a normal physique. It is essential to pregnant women. In addition, calcium is not produced in the body, so the lack of external intake of calcium is consumed in the bones may cause osteoporosis, and the amount of calcium dissolved in the blood may be unstable. Meanwhile, calcium intake can lower serum and tissue cholesterol and triglyceride levels (Calson et al., Atherosclerosis, 14, 391-400, 1971), and has recently been shown to be effective in lowering blood pressure in hypertensive patients. (Kynast-Gales and Massey, J. Am. Diet Assoc., 92, 1497-1501, 1992). Therefore, ingestion of a suitable amount of calcium is important, and has been ingesting foods and calcium-enhancing agents containing a lot of calcium.

Currently, commercially available calcium fortifiers include beef bone meal, fish bone meal, oyster shell, pearl calcium, coral calcium, whey and calcium lactate, calcium gluconate and calcium carbonate. (服 夫 高高, New Food Industry, 35 (9), 13-16, 1993).

Among these raw materials, bone bone and fish bone powder are composed of calcium phosphate like human bone, and the other ingredients are similar to human bone. These bones have long been imbued with images of calcium sources, and are now used in many forms.

The type of beef bone powder used as food is produced by burning or boiling bovine bone, which is made by burning it at a high temperature of 1,200 ℃, and is called calcined beef bone powder. 우 成) Right bone powder. In the case of firing, the calcium content is high as about 40%, and the unique bitter taste of the right bone flour does not appear, so the range of use is wide. On the other hand, in the case of unbaking is characterized by a high content of bone marrow protein has a high nutritional value, but can be used as an additive in health supplements by exhibiting a unique bitter taste. However, right bone flour is not recognized among people due to the recent surge of mad cow disease, and it is necessary to secure safety in order to expand the market (MK, BIO INDUSTRY, 14 (12)). , 56-57, 1997).

On the other hand, the fish bones, which are generated as by-products during meat processing in fish processing plants, contain large amounts of calcium and are used in the form of fish bone meal. It is made of unbaked form, and in terms of safety, it is very safe compared to beef bone powder, and thus has been in the spotlight as the next generation source of calcium.

However, fish bone meal is not widely used as a source of calcium for food, which is hard to use for food because it is hard, and has a higher unit price (5-7 times higher than fishy fishy and beef bones). Because.

Therefore, in order to solve such disadvantages of conventional fish bone meal, a method of softening fish bones by dipping in acetic acid (Japanese Patent Laid-Open No. 6-319487) has been used. However, this method is complicated and the use which can be used is limited and is not widely used.

In addition, although fish bone powder (Japanese Patent Laid-Open No. 2-231059 and Japanese Patent Laid-Open No. 4-121166), which is obtained by separating fish bones from fish, has also been produced, there is a problem in that a lot of energy is required for drying moisture. In addition, there is also a fish bone product (Japanese Patent Laid-Open No. 2-231059) in the form of pasted together with the bone protein and the muscle protein attached to the bone does not have a low calcium content.

Therefore, the inventors of the present invention have studied a method for providing fish bone meal in a simple manner with high calcium content using fish residues generated in aquatic product processing plant.

As a result, it was found that fish bone meal which can be used as a calcium fortifier can be obtained from the reaction precipitate obtained by reacting proteolytic coenzyme isolated from fish intestine with a large amount of various digestive enzymes with fish residue. Furthermore, the present invention has been found to be able to produce a peptide capable of promoting absorption of calcium from the supernatant of the reaction solution.

Accordingly, it is an object of the present invention to provide a method for producing a peptide that can promote the absorption of fish bone meal and calcium that can be used as a calcium fortifying agent from fish residues generated in aquatic product processing plants.

Another object of the present invention is to provide a fish bone meal and calcium absorption promoting peptide prepared by the above method.

1 is a schematic view showing a process for preparing a fish gut-derived proteolytic coenzyme used in the present invention.

FIG. 2 is a schematic view showing a method of preparing fish bone meal and calcium absorption promoting peptides by hydrolyzing fish residue using the coenzyme prepared in FIG. 1.

Figure 3 is a graph showing the removal rate of fish meat over time in the production of fish bone meal using tuna pyloric hydrolysing proteolytic coenzyme.

4 is a graph showing the effect of preventing calcium ions and phosphate from forming and forming calcium phosphate when calcium absorption promoting peptides according to the present invention are added where calcium ions and phosphate ions are contained (●; calcium Uptake peptide, ▽; control).

In order to achieve the above object, the method for preparing a fish bone meal and calcium absorption promoting peptides according to the present invention, the protein hydrolysis derived from the residue of fish and tuna pyloric water generated as a by-product after the fillet operation in a marine product processing plant Characterized by reacting the coenzyme.

Hereinafter, the present invention will be described in detail.

In order to produce fish bone powder, the muscles or flesh attached to the fish bones must be removed. In general, the use of strong bases such as sodium hydroxide or calcium hydroxide or strong acids such as hydrochloric acid is used to remove muscles and flesh attached to bones. (7), 69 ~ 73, 1990) However, if muscles or flesh are removed in this way, it must be neutralized and cartilage, such as chondroitin sulfate and glucosamine, is contained in bones during the treatment of strong acids and strong bases. It is composed of complex sugars and calcium-binding proteins that help bone formation are broken down and removed. However, if the meat attached to the bone is removed using the enzyme as in the present invention, the enzyme can be removed only the muscles and flesh attached to the bone by the unique substrate specificity. In addition, the enzyme reacts under mild conditions, such as strong acids or strong bases, which can prevent the destruction of bone-related substances, which may be advantageous over acids or bases. Therefore, in the present invention, fish bone meal is prepared using fish gut-derived protein, especially tuna pyloric-derived proteolytic coenzyme.

Fish bone meal prepared according to the method of the present invention has a high content of minerals such as calcium and protein, and unlike beef bone meal, there is no big problem in terms of food safety, so it can be used as a raw material of food, drink or pharmaceutical as a source of calcium, Calcium absorption accelerator peptides can be used with fish bone meal or as a source of food, beverages or pharmaceuticals for promoting calcium absorption in the form of other calcium sources or alone.

Hereinafter, the present invention will be described in detail with reference to Examples, but the present invention is not limited only to these Examples.

The fish body used as a material in the following example is a residue after fish fillet treatment in an aquatic product processing plant, and was stored in a freezer at -30 ° C to prepare fish bone meal and calcium absorption promoting peptides. In addition, the protein hydrolysing coenzyme derived from fish gut used to decompose the fish and bones was prepared by obtaining fish gut (tuna, mackerel, etc.) produced as a by-product in the seafood processing plant.

Example 1 Extraction of Fish Visceral Proteolytic Coenzyme

The method of extracting proteolytic coenzyme from the intestines of fish is as shown in FIG. In other words, add 2 times of Tris-HCl buffer solution (pH 7.0) to finely chopped fish intestine (2 kg) and homogenize it twice with a homogenizer (Ace homogenizer, Nessei AM-7) (10,000 × rpm, 2 minutes) Then, it was activated by reacting for 3 hours in a 40 ℃ constant temperature water bath. The reaction mixture containing the activated enzyme solution was centrifuged (8,000 × rpm, 20min, 4 ° C.) and the same amount of acetone was added to the supernatant and left for 6 hours in a low temperature room (4 ° C.) to obtain a protein precipitate. Centrifugation (2,370 × g, 10 minutes) was added again to the precipitate obtained by centrifugation (2,370 × g, 10 minutes), followed by centrifugation (2,370 × g, 10 minutes), and the supernatant was lyophilized. The digested coenzyme was prepared, and stored in frozen (-30 ℃) was used in the experiment. In the present invention, the proteolytic coenzyme extracted from the tuna rye water by the above method was used.

Example 2 Preparation of Fish Bone from Fish Body Using Fish Enzyme-derived Coenzyme

The method for producing fish bone meal by removing the fish meat protein of the fish by using a tuna pyloric hydrolysis proteolytic coenzyme is as follows. That is, the raw fish body is placed in a thermostat containing 0.1 M sodium acetate buffer solution (pH 10), and the tuna pylori hydrolyzate-proteolytic coenzyme is added so that the ratio of the fish body weight is 1: 100 to 300 (w / w). . Then, the reaction temperature was set at 50 ° C. for 1 to 24 hours to hydrolyze, and then heated at 100 ° C. for 10 minutes to terminate the reaction. The reaction temperature and the enzyme reaction termination temperature are the general reaction conditions of the tuna pyloric-derived proteolytic coenzyme.

On the other hand, as can be seen in Figure 3, about 73% of fish meat was removed in 6 hours after the reaction between the body and the enzyme, nearly 24% after 24 hours to remove the fish meat can provide a complete form of fish bones , React the body and enzyme within 1 to 24 hours.

When the reaction was completed, the reaction solution was separated into a supernatant and a precipitate. The supernatant was stored separately and used for the preparation of the calcium absorption promoting peptide of the next step, and the precipitate was washed with water sufficiently and dried at room temperature.

Then, using a hot air dryer at a temperature of 80 ~ 95 ℃ again to prevent the protein in the bones denatured, using a grinder to grind the size of the particles to ?? 425㎛ (mesh No. 40) fish bone meal Got. About 100 g of fish bone meal was obtained per kg of fish. On the other hand, the size of the particles in the production of fish bone powder is properly selected and pulverized by those skilled in the art.

Example 3 Preparation of Calcium Absorption Promoting Peptides Using Fish Enzyme-derived Coenzyme

The supernatant separated from the reaction solution of Example 2 was filtered to filter out remaining fish meat or scales and the remaining residues to form a clear solution.

Then, using a ultrafiltration membrane device (Minitan II Ultrafiltration system, Millipore Co.) collected only the hydrolyzate (hereinafter referred to as "calcium absorption accelerator peptide") of the molecular weight of 10,000 Da or less, and used as a calcium absorption accelerator. .

Test Example 1 Analysis of Components of Fish Bone Powder

The general component and the mineral content of the fish bone meal obtained in Example 2 are as shown in Table 1 and Table 2 below.

Common Ingredients of Fish Bone Powder Analysis test item Bone meal moisture 5.9 Ash 64.4 protein 26.5 Party 0.4 Fat 2.8 Sum (%) 100.0

Mineral content in fish bone meal (per 100 g fish bone meal) Mineral Content (g) calcium 22.93 sign 10.22 magnesium 0.34 potassium 0.07 iron 0.0054

From Tables 1 and 2, fish bone meal prepared according to the method of the present invention contains 64.4% of ash, which can be used as a mineral supplement, and also contains 26.5% of protein bound to fish bones as a protein source. Can be used. In addition, the calcium content is also high as about 23% can be used as a calcium source.

Test Example 2 Measurement of Calcium Absorption Promotion Effect

For the calcium absorption promoting peptides obtained in Example 3, the effects of calcium absorption promoting in vitro were examined with time. That is, in general, calcium ions form calcium phosphate precipitates with phosphate ions to reduce the calcium concentration in the supernatant. If a substance that binds calcium ions with higher efficiency than phosphate ions is added, Is increased, and because it is not precipitated but becomes water-soluble, the absorption rate in the body is increased (Fugita et al., Osteoporosis International., Suppl. 1, S159-162, 1993). Therefore, calcium phosphate precipitation inhibition experiment was conducted in accordance with the method of 內藤 博 (日本 榮 養 食糧 學會 誌, 39, 433 ~ 439, 1986).

In the experimental method, 40 μg of the sample was added to 1 ml of 20 mM sodium hydrogen phosphate (Na ₂ HPO ₄ ), and 0.25 ml of 20 mM calcium chloride was added thereto and reacted at 37 ° C. for 0 to 12 hours, followed by centrifugation (1,500 × g). , 5min), and the free calcium concentration in the supernatant was measured by the inductively coupled plasma (ICP) spectrophotometer (SPS 1200A plasmaspectrometer SII) and expressed as the calcium content remaining in the supernatant. Solubilized calcium ratio (%) in the supernatant was calculated according to the following equation (1). The results are shown in FIG.

From Fig. 4, in the control (▽) to which nothing was added, the calcium concentration in the supernatant was reduced to 0.8 mM after 2 hours, whereas the calcium absorption promoting peptides of the present invention were added to the place containing phosphate ion and calcium ion. In the case of (●), the calcium concentration in the supernatant was maintained at 1.3 mM, which is about 1.5 times higher than that of the control, indicating that the calcium solubilization rate was high. This is because the calcium ion and the calcium absorption promoting peptides bind with high efficiency to prevent the formation of calcium phosphate precipitation, so that the absorption rate is increased as it prevents the formation of calcium phosphate precipitation.

As described above, the fish bone meal prepared according to the present invention has a unique taste and aroma unique to fish, has a high content of minerals and proteins including calcium, and unlike the bone bone, there is no big problem in terms of food safety. And as a protein source. In addition, the calcium absorption promoting peptide also has a significant effect in inhibiting the production of calcium phosphate is expected to act as a calcium absorption accelerator, and, in addition, synergistic effect on the supply and absorption of calcium in the body when ingested fish bone meal and calcium absorption accelerator You can expect

Claims (5)

A method of preparing fish bone meal and calcium absorption promoting peptides by reacting the remaining fish residues with proteolytic coenzymes extracted from tuna pyloric water after the fillet treatment in an aquatic product processing plant. The method of claim 1, wherein the fish bone meal is prepared by reacting a fish residue with a proteolytic coenzyme extracted from tuna pyloric water and washing, deodorizing, drying, and grinding the reaction precipitate. The method according to claim 1, wherein the calcium absorption-promoting peptide is reacted with a protein hydrolysing coenzyme extracted from tuna pyloric water to the fish residue, the reaction supernatant is filtered, and the filtrate obtained by ultrafiltration again to collect a fraction having a molecular weight of 10,000 Da or less Next, the method characterized in that it is prepared by concentration and lyophilization. Fish bone powder prepared by the method of claim 2. Calcium absorption promoting peptides prepared by the method of claim 3.