KR20100005470A - Composition containing collagen hydrolysate from marine organism and method of producing the same - Google Patents

Abstract

PURPOSE: Marine organism-derived collagen hydrolysates and a manufacturing method thereof are provided to offer various uses in many fields such as food, beauty, and pharmacy etc, and to offer good immunity from diseases. CONSTITUTION: Marine organism-derived collagen hydrolysates include 9% or more of hydroxyproline and hydroxylysine among the whole amino acid of hydrolysates. The collagen hydrolysates is originated from shell or scales of fishes. The fish is globefish or a sea bream. The average molecular weight of the collagen hydrolysates is 500 ~ 1500 peptide. The protease is added in the extracted collagen protein components.

Description

Composition containing Collagen Hydrolysate from Marine Organism and Method of Producing the Same

The present invention relates to a collagen hydrolyzate having a high content of hydroxyproline (Hydroxyproline) and hydroxylysine (Hydroxylysine) and a preparation method thereof. More specifically, the present invention relates to a collagen hydrolyzate using collagen protein components extracted from the shells or scales of pufferfish or sea bream, and a preparation method including a unique pretreatment step for preparing the same, and a filtration and purification step.

In general, collagen is a fibrous light protein that connects cells between human and animal bodies, and is widely distributed in connective and cartilage tissues of the body, and three amino acid residues of glycine, proline, and hydroxyproline are twisted together. Consists of a helix structure. Collagen having such a structure has functions such as intercellular connectivity, activation of cell functions, cell proliferation, hemostasis, and immunity, and is a protein that is required for maintaining and forming the body.

Currently, collagen is used as a raw material or an additive in various fields such as beauty, health drinks, health foods, medicines and cosmetics, and the applied industrial fields are increasing.

Prior art has been used to prepare functional food additives and cosmetic raw materials using bone or skin of bovine or similar ruminants when producing protein hydrolysates or gelatin or collagen. However, in response to the outbreak of BSE, the Korean Food Safety Authority (KFDA), the European Food Security Administration (EFSA), the US Food Security Administration (FDA), and Japan's Ministry of Health, Labor and Welfare have imported raw materials from countries with a high risk of developing mad cow disease. It is forbidden to import, import or use. In addition, protein hydrolysates using livestock such as pigs and chickens are gradually prohibited from use due to the development of swine fever or patho-genic avian influenza. Therefore, there is an active development or research on related technologies for safe natural marine organisms or plant-derived collagen or protein hydrolysates without the risk of mad cow disease, swine cholera or avian influenza.

However, protein hydrolysates and collagen derived from marine organisms are poor in quality, such as poor transparency due to fatty substances or protein denaturation temperature, compared to collagen derived from cow skin and pork skin, making them difficult to use in food, cosmetics, or pharmaceutical materials. It is known.

In addition, the fish is easily rot or rancid due to the digestion of microorganisms due to the large amount of bacteria or fungi that cause decay. In the process, a low molecular weight ammonium-based volatile component occurs, resulting in deterioration of safety or quality of the product during the production of collagen or protein hydrolyzate.

An object of one embodiment of the present invention is to provide a collagen hydrolyzate having a high content of hydroxyproline and hydroxylysine.

It is an object of another embodiment of the present invention to provide a collagen hydrolyzate derived from blowfish or sea bream.

It is an object of another embodiment of the present invention to provide a method for preparing collagen peptides including an efficient raw material pretreatment step and filtration and purification steps.

The collagen hydrolyzate according to the present invention is characterized in that the content of hydroxyproline (Hydroxyproline) and hydroxylysine (Hydroxylysine) is 9% or more of the total amino acids of the hydrolyzate.

In addition, the method for producing a collagen peptide according to the present invention, by treating the basic solution containing NaOH, Na ₂ SO ₄ and NaHCO ₃ or acidic solution containing HCl and H ₂ SO ₄ , to remove impurities from fish-derived raw materials And a pretreatment step of extracting the collagen protein component, a hydrolysis step of treating proteolytic enzymes on the extracted collagen protein component, and filtering the raw material using a filtration filter and activated carbon to remove the color and smell peculiar to the fish. It characterized in that it comprises a filtration and purification step.

The collagen hydrolyzate according to the present invention has a relatively high content of hydroxyproline (Hydroxyproline) and hydroxylysine (Hydroxylysine), and can be used in various fields such as food, beauty and medicine, as well as mad cow disease, foot and mouth disease or algae. It has the advantage of being safe against various animal-reactive diseases such as the flu.

In addition, the method for producing a collagen peptide according to the present invention, it is possible to produce a high purity collagen peptide through a unique pretreatment step, it is possible to produce a collagen peptide of low molecular weight through an environmentally friendly hydrolysis step and sequential purification step. .

The present invention relates to a collagen hydrolyzate, and more particularly, to a collagen hydrolyzate having a high content of hydroxyproline and hydroxylysine. The content of the hydroxyproline (Hydroxyproline) and hydroxylysine (Hydroxylysine) is preferably 9% or more, more preferably 10% or more, and most preferably 11% or more of the total amino acids contained in the hydrolyzate.

The hydroxyproline (Hydroxyproline) and hydroxylysine (Hydroxylysine) is a quality indicator component of marine collagen hydrolyzate, the content of which compared to the animal-derived collagen hydrolyzate, of course, the content of 7% or less of the existing Compared to the marine collagen hydrolyzate from seawater is improved.

In one embodiment, the collagen hydrolyzate is derived from shells or scales of fish, preferably from shells or from scales of blowfish or bream. This is characterized by a very strong resistance to moisturizing effect and heat denaturation temperature due to the unique protective colloid properties derived from fish.

In addition, the average molecular weight of the collagen peptide is preferably about 500 to 1,500 because the collagen peptide has an advantage of excellent digestive absorption and skin permeability due to the low molecular weight of the collagen peptide.

The collagen hydrolyzate is characterized in that it is nontoxic, tasteless and odorless. Significantly reduced discoloration and release of septic, which has been pointed out as a problem of collagen hydrolyzate extracted from conventional fish.

The present invention also provides a method for preparing collagen peptides derived from fish.

In one embodiment, the manufacturing method, pretreatment of the raw material to remove impurities from the fish-derived raw material, extracting the protein, hydrolysis step of treating the proteolytic enzyme to the extracted protein component, and the filter and activated carbon Filtration and purification steps to filter the raw materials and remove fish's unique color and odor.

In the pretreatment step of the raw material, it is preferable to treat the basic solution containing NaOH, Na ₂ SO ₄ and NaHCO ₃ or an acidic solution containing HCl and H ₂ SO ₄ . In addition, the concentration of the basic solution is in the range of 0.1 to 2.0 N, the concentration of the acidic solution may be freely selected in the range of 0.1 to 2.0 N. This concentration range is the optimal concentration range through a variety of repeated experiments.

In some cases, after the pretreatment of the raw material, a sterilization step may be added to remove the decay-causing bacteria remaining in the raw material by atmospheric pressure and heating (60 to 90 ° C.) for 0.5 to 2 hours.

In one embodiment, in the hydrolysis step, the hydrolase is composed of a single form consisting of an endo type protease (eg, an alkalase), or a mixture (eg, flaverzyme) of an endo type protease and an exo type peptidase. It may be a complex form. In another embodiment, the content of the hydrolase is 0.5 to 5% by weight of the raw material, preferably 2 to 2.5% by weight of the raw material. The content of the hydrolase is not particularly limited, but is for improving the hydrolysis efficiency.

The filtration and purification step is to limit the molecular weight of the collagen peptide to a certain range, that is, the range of 500 to 5,000, and is to remove the color and odor peculiar to fish. Basically, the filter and the activated carbon is filtered and purified, and may be classified or added in the following detailed steps as necessary.

Filtration step of filtering insoluble protein and undigested raw materials using a sieving device of 20 to 100 mesh and a diatomaceous earth filter with 5 to 10% of diatomaceous earth per raw material weight, 0.5 to 1 micro filter Microfilter filtration step to remove fine deposit using system, Decolorization and deodorization step to remove color and odor by adding 5-10% of silicon and activated carbon of natural origin per raw material weight, Ultra filter system and Nano filter system Ultrafiltration and nanofilter purification step of selectively decomposing the polymer polypeptide component of the hydrolyzate using sequentially, and desalting step of desalting the hydrolyzate using the electrolysis and ion exchange resin methods sequentially may be applied. . The above detailed steps may be partially added or omitted depending on the purpose of use or need.

The method preferably further comprises a drying and powdering step of drying and powdering the sterilized residual material after the filtration and purification step. The drying and pulverizing step may be powdered, for example, using a spray dryer having an evaporation amount of 250 L / hr of an automizer type. In addition, after the selection using the automatic sieve (sieve) device of 100 to 120 mesh may include the step of packaging.

Hereinafter, the present invention will be described in more detail through preferred embodiments of the present invention, but the following examples and the like are intended to confirm the effects of the present invention by way of example, and the scope of the present invention is not limited thereto.

Example 1 Preparation of Blowfish-derived Collagen Peptides

After preparing the shell of the pufferfish, a marine by-product, the basic mixed solution of NaOH, Na ₂ SO ₄ and NaHCO ₃ corresponding to 1 N, and the acid mixed solution of HCl and H ₂ SO ₄ corresponding to 1 N in sequence Treatment to remove impurities. Then, a blowfish shell sample of high protein purity was prepared through a sterilization process using atmospheric pressure heating.

After the pretreatment step, the puffer shell samples were treated with proteolytic enzymes. The treated hydrolase is a complex form composed of endo-type protease and exo-type peptidase (flavorzyme), and treated with 2% by weight of the raw material.

After the hydrolysis step, the puffer-derived collagen peptide was prepared by sequentially decolorizing and deodorizing, and purification processes such as Micro, Ultra, and Nano Filter system.

Example 2 Preparation of Sea Bream-derived Collagen Peptides

Compared to Example 1, collagen peptides were prepared in the same manner, except that sea bream was used as a raw material instead of puffer fish.

Comparative Example 1 Preparation of Collagen Peptides from Fish

Protein components were extracted by acid hydrolysis of fish scales, skins and bones. Then, using activated charcoal, decolorization and deodorization to prepare fish-derived collagen peptide.

Experimental Examples 1 to 6 Measurement of Collagen Peptide Content According to Base and Acid Concentration

Experimental Example 1

In this experimental example, in the raw material pretreatment step of Example 1, the base treated with the raw material was selected as NaOH. The content of collagen peptide was measured while adjusting the concentration of NaOH, and the results are shown in the following table.

NaOH concentration (N) 0.05 0.25 0.5 0.75 One 1.5 2 Collagen Peptide Content (BRIX) 4.2 5.4 7.4 8.2 9.2 8.8 7.2

Experimental Example 2

Hereinafter, Experimental Examples 2 to 6 were conducted in the same manner as in Experimental Example 1, except that the type of base or acid to be treated was changed.

Na ₂ SO ₄ concentration (N) 0.05 0.25 0.5 0.75 One 1.5 2 Collagen Peptide Content (BRIX) 4.8 4.8 7.4 7.4 7.4 7.2 7.2

Experimental Example 3

NaHCO ₃ concentration (N) 0.05 0.25 0.5 0.75 One 1.5 2 Collagen Peptide Content (BRIX) 5.8 5.8 6.2 7.2 7.4 7.2 4.8

Experimental Example 4

HCl concentration (N) 0.05 0.25 0.5 0.75 One 1.5 2 Collagen Peptide Content (BRIX) 6.4 7.4 8.8 9.2 9.2 8.4 8.2

Experimental Example 5

CH ₃ COOH concentration (N) 0.05 0.25 0.5 0.75 One 1.5 2 Collagen Peptide Content (BRIX) 2.2 3.4 3.8 5.0 5.8 6.2 6.2

Experimental Example 6

H ₂ SO ₄ concentration (N) 0.05 0.25 0.5 0.75 One 1.5 2 Collagen Peptide Content (BRIX) 4.2 4.4 5.8 6.4 6.8 4.2 4.2

Combining the results of Experimental Examples 1 to 6, it can be seen that the concentration of the base and acid to be treated is relatively increased in the content of the peptide extracted in the range of 0.1 to 2.0 N.

Experimental Examples 7 to 8 Measurement of the content of collagen peptide according to the formulation of the optimum concentration.

Experimental Example 7

Experimental Examples 7 and 8 were prepared in the same manner as in Example 1, except that the concentrations of the basic solution and the acidic solution treated in the pretreatment step of the raw materials were sequentially different. The content of the prepared collagen peptide is measured and shown in Tables 7 and 8, respectively.

NaOH + NaHCO ₃ + Na ₂ SO ₄ Concentration (N) 0.05 0.25 0.5 0.75 One 1.5 2 Collagen Peptide Content (BRIX) 5.4 6.8 8.2 9.4 9.6 9.2 74

Experimental Example 8

HCl + H ₂ SO ₄ concentration (N) 0.05 0.25 0.5 0.75 One 1.5 2 Collagen Peptide Content (BRIX) 6.2 6.4 8.2 9.2 9.6 8.4 6.2

Comparative results of Experimental Examples 7 and 8 and Experimental Examples 1 to 6 can be confirmed through the following FIG. 1. Referring to Figure 1, it can be seen that the content of collagen was increased at a certain concentration, for example, 0.1 to 2.0 N range. In addition, it can be confirmed that more efficient treatment is possible when various kinds of bases or acids are properly blended as compared with the case of treating the base or the acid alone.

Experimental Examples 9 to 10 Measurement of collagen peptide content and hydrolysis rate according to the type or mixture of proteolytic enzymes.

Experimental Example 9

Compared to Example 1, except that the type of enzyme treated in the hydrolysis step was different, collagen peptides were prepared in the same manner. Hydrolysis rate and reaction conditions of the prepared collagen peptide are shown in Table 9 below.

pepsin Papain Endo-type protease + exo-type peptidase Endo-type protease Trypsin Protease Temperature (℃) 30-40 50-60 45-55 55-65 65-75 50-60 pH 2 ~ 3 5 ~ 6 5.5 ~ 6.5 6 ~ 7 8-9 6 ~ 7 Response time (hr) 60-72 36-48 24-36 24-48 60-72 36-48 Collagen Peptide Content BRIX) 6.4 8.6 8.8 9.4 6.8 9.2 Hydrolysis Rate (%) 60 80 85 90 65 85

 Experimental Example 10 Comparison of Molecular Weight of Collagen Peptides.

The molecular weight distribution of the blowfish and sea bream-derived collagen peptides according to Examples 1 and 2 and the conventional fish-derived collagen peptides according to the above 1 were compared. Molecular weight distribution was measured for each analytical sample using high performance liquid chromatography (HPLC). The results are shown in Figures 3, 4 and 5, respectively, summarized in Table 10 below.

Molecular Weight Molecular weight distribution (%) by raw material type Blowfish-derived collagen peptide Bream-derived collagen peptide Traditional Fish-derived Collagen Peptides 500 or less 29.2 43.4 22.4 500-1,000 50.8 44.2 42.4 1,000-5,000 19.8 12.4 34.5 5,000-10,000 0.2 0 0.7 More than 10,000 0 0 0 Weight average molecular weight 757 604 969

3 to 5 and Table 10, the molecular weight distribution of the blowfish or sea bream-derived collagen peptide according to the present invention compared to the conventional fish-derived collagen peptide is distributed in a relatively low range and the weight average molecular weight is significantly low It can be seen that.

Experimental Example 11 Molecular Weight Analysis of Blowfish-Derived Collagen Peptides.

The molecular weight distribution of the blowfish-derived collagen peptide according to Example 1 was analyzed by mass spectrometry (MS). The analysis results are shown in FIG. 6, which is summarized in Table 11 below.

Molecular Weight Molecular Weight Distribution (%) 250 or less 0 250-500 25.8 500-750 18.2 750-1,000 33.7 1,000-1,250 8.9 1,250-1,500 13.4 1,5000 or more 0 Weight average molecular weight 650 ~ 850

Referring to Figure 6 and Table 11, it can be seen that the molecular weight of the blowfish-derived collagen peptide according to the present invention is mainly distributed in the range of 250 to 1,000, the weight average molecular weight is 650 to 850 conventional fish-derived collagen It can be seen that it is significantly lower than the peptide.

Experimental Example 12 Analysis Comparison of Amino Acid Composition.

The amino acid composition of the fish and collagen peptides derived from puffer fish and sea bream according to Examples 1 and 2 and the conventional fish-derived collagen peptides according to the above 1 were analyzed. For each collagen peptide, amino acid analyzer was used to determine the amino acid content. The measurement results are shown in FIG. 7 and summarized in Table 12 below.

Raw Material Type Amino Acid Amino Acid Composition (%) Blowfish-derived collagen peptide Bream-derived collagen peptide Collagen peptides derived from existing fish Aspartic acid 5.69 5.33 7.15 Throneine 2.96 2.96 3.25 Serine 3.80 3.49 3.86 Glutamic acid 10.69 10.49 10.98 Proline 11.72 11.37 7.94 Glycine 22.94 22.68 17.83 Alanine 10.03 10.93 9.83 Valine 2.48 2.51 3.43 Isoleucine 1.19 1.25 2.02 Leucine 2.63 2.35 3.88 Tyrosine 0.74 0.68 0.89 Phenylalanine 1.76 1.89 1.75 Histidine 1.27 1.82 1.39 Lysine 3.30 3.49 4.87 Arginine 7.27 6.89 6.84 Cystine 0.12 0.11 0.52 Methionine 1.49 2.16 2.99 Hydroxy-proline 10.58 8.2 6.53 Hydroxy-Lysine 1.88 1.09 0.99

7 and Table 12, the blowfish or sea bream-derived collagen peptide according to the present invention compared to the conventional fish-derived collagen peptide, the content of proline and glycine increased, especially the content of hydroxyproline and hydroxylysine It can be seen that this is increased.

Figure 1 is a graph showing the results of comparing the measurement of collagen peptide content according to the pretreatment of the basic solution or acidic solution by concentration.

Figure 2 is a graph showing the results of comparing and measuring the change in the content of collagen peptide according to the type of hydrolase.

3 to 5 show the results of measuring the molecular weight distribution using high performance liquid chromatography (HPLC) for the blowfish-derived collagen peptide, the sea bream-derived collagen peptide, and the conventional fish-derived collagen peptide.

Figure 6 is a graph showing the results of measuring the molecular weight distribution of the blowfish-derived collagen peptide using a mass spectrometer (MS).

7 is a result of comparing and analyzing the amino acid composition of the collagen peptide.

Claims (10)

A collagen hydrolyzate, characterized in that the content of hydroxyproline (Hydroxyproline) and hydroxylysine (Hydroxylysine) is at least 9% of the total amino acids of the hydrolyzate. The collagen hydrolyzate of claim 1, wherein the collagen hydrolyzate is derived from the shell or scales of the fish. The collagen hydrolyzate according to claim 2, wherein the fish is blowfish or sea bream. The collagen hydrolyzate of claim 1, wherein the collagen hydrolyzate comprises a peptide having an average molecular weight of 500 to 1,500. Pretreatment of raw materials for removing impurities from fish-derived raw materials and extracting collagen protein components by treating basic solutions containing NaOH, Na ₂ SO ₄ and NaHCO ₃ or acidic solutions containing HCl and H ₂ SO ₄ , A hydrolysis step of treating proteolytic enzyme to the extracted collagen protein component, and Method of producing a collagen peptide comprising a filtration and purification step of filtering the raw material using a filtration filter and activated carbon to remove the color and smell peculiar to the fish. The method of claim 5, wherein The hydrolase is a single form consisting of an endo-type protease, or a method for producing a collagen peptide, characterized in that the complex form consisting of a mixture of endo-type protease and exo-type peptidase. The method of claim 6, The amount of the hydrolase is, the method of producing a collagen peptide, characterized in that 2 to 2.5% by weight of the raw material. The method of claim 5, wherein The filtration and purification steps include a micro filter filtration step of removing fine deposits using a micro filter system, a decolorization and deodorization step of removing color and odor by adding silicon and activated carbon of natural origin, an ultra filter system and a nano filter. Method for producing a collagen peptide comprising the step of using a system to sequentially and ultra-filter and nano-filter purification to selectively decompose the polymer polypeptide component of the hydrolyzate. The method of claim 8, The filtration and purification step may further include a desalting step of desalting hydrolyzate using an electrolysis method and an ion exchange resin method sequentially after the ultrafilter and nanofilter purification steps. The method of claim 5, wherein The method, the method of producing a collagen peptide further comprises a powder drying step of drying and powdering the remaining material after the filtration and purification step.

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