JP2005343851A - Peptide derived from fishes and method for producing the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a peptide characterized as comprising desmosine and isodesmosine which are amino acids contained only in elastin at a high concentration and definitely discriminated from collagen peptide. <P>SOLUTION: The peptide obtained by degrading a tissue composed of elastic fibers of fishes with a high elastin content, especially an aortic bulb with an enzyme, an acid or an alkali or its aqueous solution is provided. Furthermore, a method for producing the peptide comprising degrading the tissue composed of the elastic fibers of the fishes with the high elastin content with the enzyme, acid or alkali. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to a degradation product (peptide) that is obtained from a tissue composed of elastic fibers of fish, in particular, a heart arterial sphere or other arterial tissue, and has no fish-specific taste and odor, and a method for producing the same. This degradation product contains soluble elastin peptides useful as various medical biomaterials, cosmetic materials, health food materials, and the like at high concentrations.

  Elastin is present in animal tissues such as the ligaments and arteries that always require elasticity and elasticity. Moreover, although it is slight, it exists in dermis connective tissue with collagen as a component which is concerned with elasticity. Elastin in the skin is reduced by aging and ultraviolet rays, and elastin itself is denatured and cannot maintain its original elasticity, causing wrinkles and tarmi. For this reason, elastin is effective in the cosmetics and health food fields, and is currently applied to many products.

  In addition, since elastin plays an important role as a vascular component, it is considered to be involved in various vascular lesions, and its use in the pharmaceutical field is also being studied.

  Due to its properties, elastin exhibits strong resistance to various solvents such as acids and alkalis, so it must be solubilized for use in cosmetics and foods. By reducing the molecular weight, it is considered that the solubility in water and various solvents is increased and the absorption rate in vivo is increased, and it can be expected to have physiological activity.

  As the physiological effect of elastin, the effect of external preparations can be improved by applying it to the skin and hair (Japanese Patent Laid-Open No. 60-258107), and it should be used together with collagen and chondroitin Thus, it is known that it has an effect on wound healing (Japanese Patent Laid-Open No. 60-64909). Furthermore, it is known that the effects of these three components of collagen, chondroitin, and elastin are recognized even when taken orally (Japanese Patent Laid-Open No. 2003-55255). From the background as described above, it is now actively used as a functional material in the food and drink and cosmetic fields.

  Bovine ligament is known as a high-elastin-containing tissue, but cows infected with mad cow disease have been found in Japan, and elastin derived from mammals tends to be avoided due to the occurrence of livestock plague such as swine and foot-and-mouth disease It is in. Even for collagen peptides that have already been established as health food materials, most products using marine materials have become mainstream, and the market for livestock-derived materials is gradually shrinking. The same applies to elastin peptides, and materials that replace elastin peptides made from bovine ligaments are needed.

  For the reasons as described above, a safer fish-derived elastin peptide has attracted attention.

  Methods for solubilizing elastin are described, for example, in JP-A-53-101592 and JP-A-60-258107. These methods are used in mammals such as cattle and pigs that are rich in elastin. The material is a knot ligament.

  The conventional method for obtaining elastin peptides from fish materials is to use fish skin as a raw material, and extract elastin contained in trace amounts in the fish skin together with collagen from the washed fish skin using an acidic aqueous solution. A method (JP-A-5-202097), a method of treating washed fish skin with elastase (JP-A-2001-72572), and the like are known.

  However, it is known that the skin is mainly composed of collagen fibers such as collagen and contains only a very small amount of elastin. In humans, about 80% of collagen is contained in dry skin dermis, whereas elastin is about 4%.

  Similarly, the main component of fish skin is also collagen fibers, and it is recognized from the amino acid composition that the main component of fish skin is almost collagen. Furthermore, it contains almost no desmosine or isodesmosine which proves that it contains elastin. From the above, fish skin is unsuitable as a raw material for obtaining a high concentration of elastin derived from fish.

  In contrast, arterial spheres, whose main component is elastic fibers, have the characteristics of elastin protein, and the contents of desmosine and isodesmosine are almost the same as those contained in normal fish elastin.

  The object of the present invention is to use fish-derived elastic fiber as a raw material, it contains desmosine and isodesmosine, which are amino acids peculiar to elastin, and can be suitably used for health foods and cosmetics, and has a fish-specific taste and smell. It is an object of the present invention to provide a fish elastic fiber-derived peptide having a low level of safety and a method for producing the same.

  ADVANTAGE OF THE INVENTION According to this invention, the peptide obtained by decomposing | disassembling the structure | tissue comprised with the elastic fiber of fish with high elastin content with an enzyme, an acid, or an alkali, or its aqueous solution is provided.

  According to the present invention, there is further provided a method for producing a peptide comprising decomposing a tissue composed of fish elastic fibers having a high elastin content with an enzyme, acid or alkali.

  In order to achieve the above-mentioned object, according to the present invention, as a tissue constituted by elastic fibers, in particular, a tissue constituted by elastic fibers containing elastin at a high concentration (usually 30 to 50% in the total protein), In particular, blood vessels around the heart, preferably arterial spheres, are used as raw materials.

  Blood, lipids, soluble proteins, and collagenous substances are removed from the above raw materials, and the recovered insoluble protein can be decomposed with an enzyme, acid, or alkali to obtain a soluble peptide.

  According to the present invention, compared to conventional fish skin-derived elastin-containing peptides mainly composed of collagen, it has an amino acid structure unique to elastin, and further, desmosine and isodesmosine, which are amino acids unique to elastin, are contained in fish skin. A peptide containing more than the amount contained in can be obtained.

  As described above, the fish-derived peptide according to the present invention has an amino acid composition unique to elastin, contains much more desmosine and isodesmosine, which are amino acids contained only in elastin, and contains 1% or less hydroxyproline, It contains lysine in an amount of 1% or less.

  The origin of the fish arterial sphere used in the present invention is not particularly limited, but a certain size is required to collect the arterial sphere from the heart. For example, skipjack, tuna, swordfish, cod, yellowtail, yellowtail, salmon Fish raw materials obtained from trout and the like can be used. Among them, arterial spheres obtained from skipjack, tuna, cod, yellowtail, and salmon are preferred as fish species that can be stably obtained in large quantities.

  The peptide of the present invention can be used as an active ingredient of cosmetics by blending in conventional cosmetic ingredients, and as an active ingredient of health foods by blending in health foods. By applying a cosmetic containing the elastic fiber-derived fish peptide of the present invention to the skin, it is expected to improve the texture of the skin, such as elasticity and water retention, and ingesting a health food containing the peptide of the present invention By doing so, the improvement of the texture of the skin and the like is also expected.

  When the peptide of the present invention is used as an active ingredient of a cosmetic, the peptide is usually used in an amount of 0.01% to 1% of the total weight of the cosmetic ingredients, and the peptide of the present invention is used as a health food. When used as an active ingredient, the peptide is usually preferably used in an amount of 1% to 5% of the total weight of the components of the health food. Both can be used at higher or lower concentrations.

  The fish-derived elastin-containing peptide of the present invention can be produced by the production method exemplified below, but is not limited thereto, and is an ordinary hard protein solubilization method such as an alkali-ethanol method, acid degradation. Laws can also be used.

Production method 1
Arterial spheres are collected from the heart, and blood is removed by washing with running water. As the pretreatment of the raw material, the tissue is immersed in an alkali in order to facilitate the cleaning of the raw material and the subsequent extraction operation. The alkali in this case is a solution of sodium hydroxide, calcium hydroxide, preferably sodium hydroxide. The concentration is 0.01N to 0.1N, preferably 0.02N. The immersion temperature is 20 ° C. or less, and the period is about 12 hours to 5 days, preferably 3 days. During immersion, it is preferable to change the solvent at least twice a day. The conditions for these pretreatments are not limited to this because the resistance of the raw material tissue to the solvent differs depending on the fish species, and it is desirable to examine in advance depending on the raw fish species.

  After soaking, excess alkali is removed by washing with running water to neutralize the raw material. After washing, distilled water is added thereto and heated to 95 ° C. to remove collagenous substances and lipids. This is enzymatically decomposed as a raw material for extraction to obtain soluble elastin. Enzymes used for degradation can be used as long as they can be used in the food, pharmaceutical and cosmetic industries, but those having a high titer are preferable, such as Alcalase 2.4L FG (Nnovozymes), Protin AC-10F ( Daiwa Kasei), protease N “Amano” G, pepsin (manufactured by Amano Enzyme) and the like are desirable, and the degradation conditions are determined for each of these enzymes used. The amount added is usually an amount used for proteolysis in the art, for example, the weight ratio of enzyme to substrate is 1: 5000 to 1: 10,000. These enzymes are preferably used in combination of two types rather than using them alone. After completion of the reaction, heat inactivation is performed, and the pH is adjusted to about 4 which is a preferable pH as a cosmetic raw material, and then the residue is separated by filtration. At this time, if necessary, the insoluble matter is removed using activated carbon, a filter aid or the like to obtain an elastin-containing peptide solution. When using it as a solution, sterilization filtration is performed using a membrane filter here.

  The enzyme degradation product is further concentrated and spray-dried, so that it can be used not only as a cosmetic raw material but also as a health food raw material.

Manufacturing method 2
In the same manner as in Production Method 1, a raw material pretreated with an alkali is used and heated in an inorganic acid solution to obtain an acid-soluble elastin peptide. As the acid to be used, hydrochloric acid and oxalic acid are preferable. Treatment with 1N, 80 ° C. for hydrochloric acid and 0.25 M, 90 ° C. for oxalic acid is preferable. After solubilization, neutralization with alkali is performed. As the alkali used at this time, sodium hydroxide and calcium hydroxide are preferable. In particular, when solubilized with oxalic acid, neutralization with calcium hydroxide is indispensable in order to completely remove it. . Residues and generated salts are removed by filtration, and desalting is performed if necessary to obtain an elastin-containing peptide solution.

Production method 3
In the same manner as in Production Method 1, using raw materials pretreated with alkali, solubilize in 1N-NaOH, 80% EtOH solution at room temperature. After removing the residue by filtration, the pH is adjusted to around 4, and if necessary, desalting is performed to obtain an elastin-containing peptide solution.

  Examples of the present invention are shown below.

Example 1
100 g of fresh bonito heart arterial spheres are washed with distilled water and the blood inside is removed. The washed arterial sphere is immersed in 0.02N sodium hydroxide solution at room temperature for 3 days to remove the remaining blood. After washing with running water, neutralize with hydrochloric acid, add 3 volumes of distilled water, and sterilize by heating at 95 ° C. Remove the supernatant and remove the collagen along with the lipid. Using this as a raw material for extraction, it is fragmented with a food cutter, added with 1.0% of enzyme Alcalase 2.4L FG, and decomposed in a solution adjusted to pH 8.5 with sodium hydroxide for 6 hours. Add hydrochloric acid to the decomposition product to pH 3, add pepsin (Amano Enzyme) to 1: 10,000, and decompose for 16 hours. Adjust the pH to 4, inactivate the enzyme at a temperature not lower than 85 ° C, purify by layer filtration and then spray dry to obtain 12 g of soluble peptide powder.

Example 2
100 g of fresh Hamachi heart arterial spheres are washed with distilled water and the blood inside is removed. The washed arterial sphere is immersed in 0.01N sodium hydroxide solution at room temperature for 3 days to remove the remaining blood. After washing with running water, add 3 volumes of distilled water and heat sterilize at 95 ° C. Remove the supernatant and remove the collagen along with the lipid. Using this as a raw material for extraction, shred it with a food cutter, add 0.5% of the enzyme protin AC-10F (Daiwa Kasei), decompose for 2 hours, and add 0.1% of the enzyme protease N “Amano” G (Amano Enzyme) And decompose for 5 hours. The enzyme is inactivated at a temperature of 85 ° C or higher, purified by layer filtration and spray-dried to obtain 7.3 g of soluble peptide powder.

Example 3
100 g of fresh bonito heart arterial spheres are washed with distilled water and the blood inside is removed. The washed arterial sphere is immersed in 0.02N sodium hydroxide solution at room temperature for 3 days to remove the remaining blood. After washing with running water, add 3 volumes of distilled water and heat sterilize at 95 ° C. Remove the supernatant and remove the collagen along with the lipid. Immerse in 0.2N hydrochloric acid for 5 days, remove the supernatant, add 0.1% pepsin and decompose at 37 ° C for 24 hours. The pH is adjusted to 4 with NaOH, the enzyme is inactivated at a temperature not lower than 85 ° C., purified by layer filtration and spray-dried to obtain 10 g of soluble peptide powder.

  Compared with a conventional fish skin-derived elastin peptide, an enzyme degradation product having a high content of desmosine and isodesmosine, which is a characteristic of elastin, can be obtained. It was also confirmed to have a composition different from that of fish collagen.

  The amino acid composition (mol / 1000 mol) of the enzymatic degradation product prepared according to Examples 1 and 2 is shown in the table below. For comparison, the amino acid composition of peptides extracted from tuna skin was also shown.

Claims (4)

  A peptide obtained by decomposing a tissue composed of fish elastic fibers having a high elastin content with an enzyme, acid or alkali, or an aqueous solution thereof.   The peptide according to claim 1 or an aqueous solution thereof, wherein the tissue composed of elastic fibers of fish is a fish arterial sphere or other arterial tissue.   The fish-derived peptide or an aqueous solution thereof according to claim 1, which has an amino acid composition unique to elastin and contains desmosine and isodesmosine, which are amino acids contained only in elastin, in a concentration higher than that contained in fish skin.   A method for producing a peptide, comprising decomposing a tissue composed of fish elastic fibers having a high elastin content with an enzyme, an acid or an alkali.