JP2010037236A - Cystine-collagen peptide composite and method for producing the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an excellent functional material having both the function of a collagen peptide and the function of cystine. <P>SOLUTION: A cystine-collagen peptide composite comprises cystine in an amount of not more than 10 times the amount of an collagen peptide. The cystine-collagen peptide composite is produced by undergoing a step of dissolving cystine in an alkali solution having a pH of 10 or more and a step of mixing and agitating the obtained cystine solution and a collagen peptide solution to form a composite in a solution having a pH of less than 10. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to adding a new property to a collagen peptide. More specifically, the present invention relates to a cystine-collagen peptide complex that imparts a sulfur-containing amino acid cystine to a collagen peptide and forms a precipitate in a pH-dependent manner. The present invention relates to a cystine-collagen peptide complex that can be used in place of a material that has been produced using collagen peptide as a raw material.

  Collagen peptides are widely recognized and accepted by consumers in the health foods and supplements market because they are relatively easy to experience the effects on skin, bones, and joints. Examples of raw materials include pigs, cattle, and fish, but the majority of porcine raw materials are collagen peptides. The collagen peptide is obtained by hydrolyzing gelatin obtained by heating and extracting skin, bone, scales, etc. with an enzyme or acid / alkali, and purifying it. It has a lower molecular weight than gelatin and does not gel at low temperatures. Collagen peptides often have a molecular weight of about several thousand, and are aggregates of peptides having a size of about 2000 to 10,000. The final product is provided in the form of dried powders, granules and solutions.

  Collagen that is the basis of collagen peptides is a fibrous protein that is contained in the largest amount in the body of multicellular animals including humans, and occupies about 25% of the total protein in the body. It is a major component of connective tissue such as skin, bone, cartilage, and tendon, and serves as a structural material that forms, holds, and reinforces the body and organs, and as a foundation for bonding cells together. 40% of the total collagen is contained in the skin, 20% is contained in bone and cartilage, and is widely distributed throughout the body including blood vessels and internal organs.

  The functionality of collagen peptides is roughly divided into physical functions and physiological functions. Physical functions include water retention, protective colloid properties, binding properties, thickening properties, and crystal precipitation suppression. Moreover, since it is both electrolytes like gelatin, it also has a pH buffering action. The physiological characteristics include a function as a nutrient and a function as a physiological activity / biological regulation function. The nutritional characteristics are basically similar to those of gelatin, and the amino acid composition of Gly (glycine) occupies 1/3 of the whole, and is characterized by being rich in imino acids such as Pro (proline) and Hyp (hydroxyproline). Have. However, there is little or no Cys (cysteine / cystine) and no essential amino acid Trp (tryptophan) (Non-patent Document 1).

  Cystine is an amino acid having a structure in which two molecules of cysteine are linked via an S—S bond (disulfide bond) generated by oxidation of a thiol group (—SH). Disulfide bonds between cysteines in polypeptides are necessary to maintain the secondary structure of proteins, and in hair, disulfide bonds between cysteines in keratin determine the degree of curly hair. There are various proteins in hair, but the main protein is called keratin and is composed of about 18 amino acids. It is characterized by containing 14 to 18% of cystine that is hardly contained in other proteins such as silk and fiber (Non-patent Document 2). Furthermore, keratin present in hair is also present in nails, and its amino acid composition is similar to that of hair and contains a large amount of cystine. A disulfide bond is formed between cysteine residues in the keratin fiber to form cystine, which contributes to the improvement of the strength of the keratin fiber.

  Therefore, in the field of cosmetics for hair, a cystine-introducing peptide to which the characteristics of cystine are added has been developed (Patent Document 1), and a hair protecting agent for preventing damaged hair and recovering damaged hair using this technique. (Patent Document 2) and the first agent for permanent wave that can prevent hair damage during permanent wave, can impart excellent wave to hair, and can give gloss and moisture to treated hair (patent Document 3) is provided. In the field of tissue engineering and biotechnology, collagen derivatives that can be cross-linked via disulfide bonds are modified by adding thiol groups for the purpose of application to biomaterials such as implants. 4) has been developed. Moreover, an organic compound having a thiol group (Patent Document 5) has been developed and provided as a suture, a biological substitute, and an adhesive. As described above, progress has been made in the development of materials that have the characteristics of cystine in the fields of hair cosmetics, tissue engineering, and biotechnology, but not in other fields such as food and drink, but the characteristics of cystine have been added. The development of such materials was expected.

  As described above, in the development of a material having both characteristics by combining two kinds of compounds, development of a modified collagen peptide in which another compound is bound to a collagen peptide is also progressing. For example, in the field of external preparation for skin, a carotenoid excellent in cell activation and antioxidant properties is given a collagen peptide excellent in biocompatibility, and the carotenoid-collagen peptide condensate (Patent Document 6) is biocompatible and stable in the preparation. It is provided for the purpose of improving. In the field of tissue engineering and biotechnology, a biocompatible cohesive polymer gel (Patent Document 7) by co-precipitation of dextran sulfate and gelatin has been developed and provided as a suture, a biological substitute, and an adhesive. As described above, progress has been made in the development of materials having the properties of collagen peptides in the fields of topical skin preparations, tissue engineering and biotechnology, but not in other fields such as foods and beverages and collagen peptides. The development of materials with the characteristics of the was expected.

  Food / beverage products / cosmetics containing sulfur-containing amino acids such as cystine, cysteine, methionine, and taurine or protein hydrolysates containing sulfur-containing amino acids with collagen peptides have been provided for the purpose of improving the hair-growth and hair-care effects. (Patent Document 8, Patent Document 9). However, there is no food / beverage product / cosmetics containing sulfur-containing amino acids or protein hydrolysates containing sulfur-containing amino acids in a form combined with collagen peptides, and the properties of sulfur-containing amino acids or protein hydrolysates containing sulfur-containing amino acids The development of materials that can be used in the fields of added foods and beverages and cosmetics has been desired.

Japanese Patent Laid-Open No. 10-287797 Japanese Patent Laid-Open No. 11-269045 Japanese Patent Laid-Open No. 11-269047 Japanese Patent Laid-Open No. 6-80935 JP-T 9-503490 JP 2007-238564 A JP 2006-506110 A Japanese Patent No. 3407004 JP 2001-321125 A Food Industry 2008-5.30 Kogyo Co., Ltd. “Development of Health and Functional Food Materials I Development of Functional Collagen Peptides and Their Effects” "Science of Wave" Japan Permanent Wave Liquid Industry Association, Shin Beauty Publishing

An object of the present invention is to provide an excellent functional material having both the function of a collagen peptide and the function of cystine and a method for producing the same.
More specifically, the present invention provides a material excellent in functional performance having a function specific to collagen peptides such as prevention of skin roughness and water retention and a function specific to cystine such as enhancement of hair and nails, and a method for producing the same. For the purpose.

  As a result of intensive studies in view of the above circumstances, the present inventors have focused on functional materials such as cystine and collagen peptide, dissolved L-cystine in an alkaline solution having a pH of 10 or higher, and then mixed with the collagen peptide solution. By stirring, a complex can be formed in some form in a solution having a pH of less than 10, and according to this method, the reagent used in the synthesis process can be removed without going through the conventional complicated chemical synthesis. Since it does not need to be purified, it can be easily added to products that require attention to safety, such as foods and beverages and cosmetics, and it can be used in forms other than solutions by collecting it as a precipitate. The present invention was completed by producing a cystine-collagen peptide complex that can be produced and providing it as a novel collagen peptide.

That is, the gist of the present invention is as follows.
(1) A cystine-collagen peptide complex comprising 10 times or less of cystine with respect to a collagen peptide,
(2) dissolving cystine in an alkaline solution having a pH of 10 or higher;
The method for producing a cystine-collagen peptide complex according to (1), which comprises a step of forming a complex in a solution having a pH of less than 10 by mixing and stirring the obtained cystine solution and collagen peptide solution,
(3) It is related with the food-drinks or cosmetics containing the cystine-collagen peptide complex as described in said (1).

  In the past, proteins were considered to be broken down into amino acids in the intestinal tract and absorbed after ingestion, not limited to collagen peptides, but di- (2) and tri- (3) as research progressed in recent years. It was also revealed that it may be absorbed as a larger oligopeptide. Therefore, by taking the cystine-collagen peptide complex of the present invention by adding it to beverages, etc., sulfur-containing amino acids or sulfur-containing amino acids that are materials for proteins constituting hair, nails, and skin such as cysteine, methionine, and cystine Ingestion of protein hydrolyzate in combination with collagen peptide is synergistic in both retention and reinforcement of keratin fibers in the epidermis by sulfur-containing amino acids and retention and reinforcement of collagen fibers in the dermis by collagen peptides. The effect can be expected, and as a result, the hair, nails, and skin can be prevented from being damaged, the damaged hair, nails, and skin can be recovered, and gloss and smoothness can be expected.

  In addition, by adding the cystine-collagen peptide complex of the present invention to cosmetics and applying it to hair, nails, and skin, the biocompatibility of the collagen peptide effectively absorbs cystine from the epithelium, and keratin fibers As a result, the hair, nails, and skin can be prevented from being damaged, and the damaged hair, nails, and skin can be recovered, and gloss and smoothness can be imparted.

  As a raw material of the “collagen peptide” used in the present invention, collagen protein prepared from animals such as pigs, cows, chickens, and fish bones, skins, muscles, etc. can be used. It can be produced by fermentation and enzymatic degradation or by bioreactor degradation. The collagen peptide that can be used in the present invention is preferably one having a low viscosity regardless of the molecular weight.

  The “cystine” used in the present invention has a structure in which two molecules of cysteine, which is a sulfur-containing amino acid, are connected via an S—S bond (disulfide bond). Used.

  In the cystine-collagen peptide complex of this invention, the said cystine should just contain 1-10 times amount (weight conversion) with respect to the said collagen peptide.

  The cystine-collagen peptide complex of the present invention having the above-described configuration can be produced by mixing an alkaline solution of cystine and a collagen peptide solution.

  The “alkali” used in the present invention can be used as an aqueous solution having a pH of 10 or more. For example, potassium carbonate / sodium carbonate can be used.

The method for producing the cystine-collagen peptide complex of the present invention comprises:
Dissolving cystine in an alkaline solution having a pH of 10 or higher;
A step of forming a complex in a solution having a pH of less than 10 by mixing and stirring the cystine solution and the collagen peptide solution.

The content of cystine in the cystine solution is preferably 2% by weight or less. Moreover, it is preferable that the cystine is heated when dissolved in the alkaline solution.
Moreover, as content of the collagen peptide in the said collagen peptide solution, 50 weight% or less is preferable. The pH of the collagen peptide solution is preferably 4-5.
The mixing of the cystine solution and the collagen peptide solution may be at normal temperature or in a heated state, but is preferably at normal temperature.
The mixing ratio of the two solutions may be adjusted so that 10 times or less, preferably 1 to 10 times the amount of cystine is mixed in terms of weight per collagen peptide.

  In addition, the obtained cystine-collagen peptide complex can be purified as a precipitate by centrifugation or the like, or purified by washing with a buffer solution such as a phosphate buffer solution or a citrate buffer solution or water.

  The cystine-collagen peptide complex obtained as described above is a functional material having both characteristics of cystine and collagen peptide, and can be used not only as a substitute for cystine and collagen peptide but also as a masking agent. Useful. It is.

  Therefore, this invention relates to the food-drinks or cosmetics containing the said cystine-collagen peptide complex. The content of the cystine-collagen peptide complex in the food / beverage product or cosmetic is not particularly limited as long as the function of the complex can be exhibited.

Moreover, it is preferable that the food / beverage products containing the cystine-collagen peptide complex of this invention contain thickening polysaccharide.
The beverage containing the cystine-collagen peptide complex of the present invention is a beverage in which sedimentation of the cystine-collagen peptide complex is suppressed by the addition of the thickening polysaccharide, and the cystine-collagen peptide complex is dispersed in the liquid. Provided that cystine and collagen peptides can be easily taken simultaneously.
The content of the thickening polysaccharide in the beverage is preferably 0.1 to 0.2% by weight, more preferably 0.05 to 0.1% by weight from the viewpoint of preventing precipitation of the cystine-collagen peptide complex. preferable.

  As said food-drinks, the drink, foodstuffs, etc. which can mix | blend a cystine-collagen peptide complex are mentioned. Examples of the cosmetics include basic cosmetics such as lotion, milky lotion, face wash, and cream, finished cosmetics such as foundation, lipstick, white powder, and nail polish, and cosmetics for hair such as hair conditioner and hair tonic.

  Next, the present invention will be described more specifically with reference to examples. However, this invention is not limited only to those Examples. In the following examples and the like, “%” indicating the concentration of a solution or the like is “% by weight” unless otherwise indicated.

Example 1
The cystine-collagen peptide complex is prepared by dissolving L-cystine in a potassium carbonate solution having a pH of 10 or higher at 90 ° C. to prepare a 2% cystine solution. This is returned to room temperature (25 ° C.), and a 50% aqueous solution of collagen peptide is gradually added thereto and stirred. As a collagen peptide solution to be used, a 50% aqueous solution (pH 4.6) of a collagen peptide derived from pork material having an average molecular weight of 3000 (Nitta Gelatin Co., Ltd.) is prepared and added to the 2% cystine solution. At room temperature (25 ° C.), 32 g of a 50% solution of the above collagen peptide (Nitta Gelatin Co., Ltd.) was gradually mixed and stirred in 5 g of a 2% cystine solution, and when the pH was lowered to 9, white turbidity was produced. . Subsequently, the reaction was continued for 30 minutes at room temperature. After the reaction was completed, 8 g of 1M phosphate buffer was added to adjust the pH to 7. The reaction solution was centrifuged at 49000 × g (20000 rpm) to remove the supernatant, the remaining supernatant and the precipitate were washed with 0.1 M phosphate buffer, and the suspension was 49000 × g (20000 rpm). The mixture was centrifuged to remove the supernatant and dried to obtain 11 mg of a cystine-collagen peptide complex precipitate.

Comparative Example As a control for Example 1, at room temperature (25 ° C.), 5 g of 2% cystine-2% potassium carbonate solution was mixed and stirred with 7 g of 1M sodium dihydrogen phosphate solution to lower the pH. White turbidity occurred. Subsequently, the mixture was allowed to react for 30 minutes at room temperature. After the reaction was completed, 7 g of 1M phosphate buffer was added to adjust the pH to 7, and 32 g of a 50% hydrolyzed collagen peptide having an average molecular weight of 3000 was gradually added. Mix and stir. The reaction solution was centrifuged at 49000 × g (20000 rpm) to remove the supernatant, the remaining supernatant and the precipitate were washed with 0.1 M phosphate buffer, and the suspension was 49000 × g (20000 rpm). ), And the supernatant was removed, followed by drying to obtain 10 mg of a cystine precipitate.

Example 2
A part of the precipitate obtained in Example 1 was taken in a Durham tube, deaerated and acid hydrolyzed at 150 ° C. for 1 hour, and then amino acid composition analysis was performed with an amino acid analyzer. An amount of 22.5 mol% was detected as half cystine (content of cystine and cysteine in the amino acid analysis). Furthermore, 18.2 mol% as glycine and 5.2 mol% as proline were detected. A part of the precipitate obtained in Example 2 as a control was taken in a Durham tube, degassed and acid-hydrolyzed at 150 ° C. for 1 hour, and then subjected to amino acid composition analysis with an amino acid analyzer. Only a trace amount of proline was detected.

  Calculated from the results obtained in Example 1 and Comparative Example, the ratio of amino acids precipitated by reacting with cystine, excluding free amino acids, was expressed as the amino acid composition of pig skin gelatin (Nitta Gelatin Co., Ltd.). It is shown in FIG. 1 as the amino acid content, together with the website gelatin laboratory “General Properties of Gelatin”). In FIG. 1, the content rate of each amino acid when the ratio of Gly (glycine) contained is 1 is shown. The ratio of Cys (cystine) in the starting collagen peptide is 0 (see Nitta Gelatin Co., Ltd. website Gelatin Laboratory, “General Properties of Gelatin”), which is not shown in FIG.

  As is apparent from FIG. 1, in the precipitate obtained in the cystine-collagen peptide reaction experiment, amino acids other than Cys (cystine) include Gly (glycine), Pro (proline), Ala (alanine), and Hyp (hydroxyproline). The ratio is high. When compared with the amino acid composition of pork skin gelatin, the amino acid composition excluding Cys (cystine) in the precipitate was almost the same as the amino acid composition of pork skin gelatin. From the above, it became clear that cystine in the precipitate formed a complex with the collagen peptide.

  In the cystine-collagen peptide reaction experiment, when a collagen peptide is added to a cystine solution having a pH of 10 or more, a precipitate containing the collagen peptide is formed, and the collagen peptide is added to the cystine containing the precipitate after adjusting to pH 7. When added, no precipitate containing the collagen peptide was formed. As described above, it was revealed that the collagen peptide detected in the precipitate formed by the reaction accompanied by the change in pH formed a complex with cystine.

Further, regarding the functionality of the cystine-collagen peptide complex obtained in Example 1 (effect on skin, hair and nails), a cystine-collagen peptide-containing beverage (corresponding to 100 mg of the cystine corresponding to 5 monitors) -Containing a collagen peptide complex) and received a tasting per day for 2 weeks. A five-step evaluation was made to answer questions (improvement items) regarding whether or not the effect on skin, hair, and nails was felt at the 3rd, 7th, 10th, and 14th days of 2 weeks. The efficacy was quantified. The results are shown in FIGS. The placebo in FIGS. 2-4 means the drink containing only a collagen peptide. The evaluation in the figure is an average value.
From the results of FIGS. 2 to 4, the cystine-collagen peptide complex of the present invention has an effect on hair and nails, which is a function specific to cystine, and an effect on the skin, which is a function specific to the collagen peptide. It was found that both can be felt

  Example 3

  Each component was mixed according to the following formulation to prepare a cosmetic (cream).

Example 4
Each component was mixed according to the following formulation to prepare a beverage.

  Further, when the criticality of the cystine content in the cystine-collagen peptide complex was tested, in an experiment in which 16 g of an excessive amount of collagen peptide (average molecular weight 3000) was added to and mixed with 100 mg of cystine, 10 mg of the collagen peptide was adsorbed. The result was obtained. As a result of the experiment, 100 mg of cystine was adsorbed to 10 mg of collagen peptide having a certain molecular weight distribution out of 16 g of the excessive amount of collagen peptide, and in this case, complexed at a ratio of collagen peptide: cystine = 1: 10. Think of it as forming a body. In a preliminary experiment, when 200 g cystine was reacted with 11 g collagen peptide (average molecular weight 3000), 50 mg collagen peptide with a certain molecular weight distribution was adsorbed to 200 mg cystine. In this case, it is considered that a complex is formed at a ratio of collagen peptide: cystine = 1: 4. From the above results, in the production method of the present invention, the amount of cystine and collagen peptide contained in the cystine-collagen peptide complex can be changed by the mixing ratio of collagen peptide and cystine, but the collagen peptide having a certain molecular weight distribution can be changed. It is considered that at most 10 times the amount of cystine is adsorbed.

  Since the cystine-collagen peptide complex of the present invention is excellent in functionality having both the functions of the collagen peptide and cystine as described above, it prevents skin itchiness and water retention, enhances hair and nails, stains・ Food and drink for the purpose of inhibiting and releasing melanin, which causes freckles, inactivating antioxidants, deactivating active oxygen, assisting in the discharge of toxic substances such as heavy metals, assisting in alcohol degradation in the liver, and improving liver function It can be suitably used as an active ingredient in products or cosmetics.

FIG. 1 is a graph showing the ratio of amino acids precipitated by reacting with cystine obtained in Example 1 and Comparative Example, and the results as well as the amino acid composition of pork skin gelatin. FIG. 2 is a graph showing the results of evaluating the effectiveness of the cystine-collagen peptide complex obtained in Example 1 and placebo on the skin. FIG. 3 is a graph showing the results of evaluating the effectiveness of the cystine-collagen peptide complex obtained in Example 1 and placebo on the hair. FIG. 4 is a graph showing the results of evaluating the effectiveness of the cystine-collagen peptide complex obtained in Example 1 and the placebo on the nails.

Claims (3)

  A cystine-collagen peptide complex comprising 10 times or less of cystine with respect to a collagen peptide. Dissolving cystine in an alkaline solution having a pH of 10 or higher;
The method for producing a cystine-collagen peptide complex according to claim 1, comprising a step of forming a complex in a solution having a pH of less than 10 by mixing and stirring the obtained cystine solution and the collagen peptide solution.   A food or drink or cosmetic containing the cystine-collagen peptide complex according to claim 1.