WO2014157854A1 - Method for isolating collagen from jellyfish by using radiation - Google Patents

Abstract

The present invention relates to a method for isolating collagen from jellyfish by using radiation. Specifically, acid-soluble collagen and atelocollagen are prepared by using a combined method of radiation irradiation and chemical material treatment on jellyfish, and thus it would be useful as a method for isolating collagen from jellyfish at low cost and in high yield.

Description

 dnd [specification

 [Name of invention]

 Separation method of geranogen from jellyfish using radiation

 The present invention relates to a method of extracting collagen from jellyfish using a radiation technique, and more specifically, to collagen from jellyfish at a low cost and high yield rate using a hybrid method of irradiation technique and chemical treatment technique. It is about how to separate.

Background Art

 Collagen is a major component of the extracellular matrix and is distributed in skin, bone and cartilage proteins. Its structure is a three-helix fibrous polymer protein. Its diameter is about 14-15, its length is about 2800, and its average molecular weight is about 300,000 Da. Collagen has a physically and biologically stable structure by covalent cross-linking between tropocollagen molecules or tropocollagen molecules, which are the basic unit molecules of fibrous proteins. The general collagen peptide structure is (Gly-XY) n, where X is proline and Y is hydroxyproline, about one third, and the other two thirds are other amino acids. It is known.

Collagen is a functional material that is widely used in food, medicine, cosmetics, and cell culture industrially. It is used as an edible casing or carrier in food, and is also used as an additive to improve the texture of sausage or ham. In addition, due to various collagen functions such as fixation of cell adhesion, induction of cell division and differentiation, thrombolytic action, memory enhancement effect, wound healing and gastric mucosa function, demand is increasing and a large amount of collagen is required. It is true. Among them, animal-derived collagen, which is most commonly used as a pharmaceutical material, has recently been exposed to high-risk groups such as animal infectious agents (mad cow disease, bird flu, infectious spongiform encephalopathy), and human collagen may be used to solve these problems. Not easy to extract one There are complex disadvantages such as low productivity, high process costs and unethical social issues. In order to supplement these problems, development of wound coating materials, drug delivery materials, artificial organ materials for regenerative medicine using biopolymers extracted from marine organisms that have no risk of cell affinity, cytotoxicity and immune response compared to animal-derived proteins. And commercialization is active. In Korea, the development of high-purity biopolymer separation and purification for low molecular weight and low molecular weight process for pharmaceuticals takes up a large portion and is currently in the prototype development stage, but there is an urgent need for the convergence of new technologies to increase price competitiveness.

 Accordingly, studies on marine organism-derived collagen include amino acid analysis, solubility measurement, denaturation temperature, and solubility of acid-soluble collagen extracted from skin and bones of jellyfish, fish, and fish such as fish and terrestrial animal collagen. A similar structure was identified with no difference. Among them, jellyfish collagen has been found to be effective for the treatment of arthritis, hypertension, bronchitis, asthma, as well as control of skin elasticity and blood circulation.

However, large-scale emergence of jellyfish due to global warming has a detrimental effect on the ecosystem, and its amount is also difficult to process. These jellyfish are limited to simple foods. In addition, until now collagen is extracted from jellyfish using acid, alkali, salt, which is a simple chemical treatment method, and the previous extraction methods by chemical treatment are environmental pollution and low yield rate. It is pointed out as a problem in practical use. Accordingly, the present inventors have attempted to develop a new method for more efficient separation of collagen from jellyfish to solve the above problems, the treatment of the existing chemicals only if the jellyfish irradiation technology and the appropriate method of chemical treatment The present invention was completed by confirming that the cost, efficiency, and yield of separating collagen are improved compared to the method. [Detailed Description of the Invention]

[Technical Problem] An object of the present invention is to provide a method for separating acid-soluble collagen from jellyfish.

 Another object of the present invention is to provide a method for producing atelo collagen, which comprises the step of treating acid-soluble collagen prepared by the above method with a protease and drying.

Technical Solution In order to solve the above problems, the present invention

 1) washing and pulverizing jellyfish;

 2) dipping the pulverized jellyfish of step 1) in an acidic solution;

 3) irradiating and stirring the solution of step 2); And

 4) It provides a method for separating acid-soluble collagen (collagen) from jellyfish comprising the step of filtering the stirring of step 3) and then dried.

 In addition, the present invention

 It provides a method for producing atelo collagen comprising the step of treating the acid-soluble collagen prepared by the above method with a protease and drying.

[Effective Effect] The use of irradiation technology and chemical treatment in jellyfish can produce collagen at low cost and high efficiency, resulting in lower costs, higher yields, and better ecosystems than conventional methods. The jellyfish can be used to prevent environmental pollution, and furthermore, it can be usefully used as a separation process technology that can be used for the raw material and biomaterial manufacturing technology of jellyfish collagen, which is the basis of tissue engineering. [Brief Description of Drawings]

 1 is a flow chart illustrating a method for separating collagen from jellyfish according to the present invention.

 Figure 2 is a diagram showing the collagen extracted from the jellyfish according to the dose of gamma rays in accordance with the present invention.

 3 is a diagram showing the weight change after the washed jellyfish analysis.

 Figure 4 is a view showing the jellyfish particle size according to the grinding time.

 5 is a view showing the yield of the acid-soluble collagen extracted from jellyfish according to the stirring time after gamma irradiation.

 6 is a diagram showing the yield of acid-soluble collagen extracted from jellyfish according to the irradiation dose of gamma rays.

 Figure 7 is a diagram showing the extraction rate of atelo collagen prepared from jellyfish according to the dose line of gamma rays.

 8 is a diagram showing the chemical properties of the acid-soluble collagen extracted from jellyfish according to the radiation dose according to the present invention.

 9 is a diagram showing the thermal properties of atelo collagen prepared from jellyfish according to the radiation dose according to the present invention.

 10 is a diagram showing the components of atelo collagen prepared from jellyfish according to the present invention for each radiation dose.

[Best form for implementation of the invention]

 Hereinafter, the present invention will be described in detail. The present invention provides a method for separating acid-soluble collagen from jellyfish comprising the following steps:

 1) washing and pulverizing jellyfish;

 2) dipping the pulverized jellyfish of step 1) in an acidic solution;

3) irradiating and stirring the solution of step 2); And _c

 5

4) drying the filtered solution of step 3).

 In the manufacturing method according to the present invention, step 1) is a step of washing and pulverizing jellyfish.

 In the manufacturing method according to the present invention, jellyfish washing and pulverization of step 1) is preferably performed in the following steps, but is not limited thereto:

 I) crushing the washed jellyfish;

 II) lyophilizing the ground jellyfish of step I); And

 III) grinding the lyophilized jellyfish of step II).

 The lyophilized jellyfish is preferably pulverized to a particle size of 100 to 3000 is not limited thereto.

In the manufacturing method according to the present invention, step 2) is a step of dipping the pulverized jellyfish in an acidic solution. ^'

The acidic solution is preferably any one selected from the group consisting of acetic acid, citric acid and formic acid, more preferably acetic acid, but is not limited thereto. ^'

 The acidic solution is preferably 0.01 M to 2.0 M concentration, more preferably 0.1 M to 1.5 M concentration, more preferably 0.3 M to 1.0 M concentration, and most preferably 0.5 M concentration, but not limited thereto. Do not.

 In the manufacturing method according to the present invention, step 3) is a step of stirring after irradiating the solution with radiation.

 The radiation is preferably gamma rays or electron beams, and more preferably gamma rays, but is not limited thereto.

 The radiation is preferably irradiated with an irradiation dose of 5 kGy to 200 kGy, more preferably an irradiation dose of 5 kGy to 100 kGy, more preferably an irradiation dose of 5 kGy to 50 kGy, and 5 kGy to 25 It is more preferable that it is the irradiation dose of kGy, It is most preferable, but it is not limited to 10 kGy.

Outside the above range, there is no effect of extracting collagen by radiation at a radiation dose of 5 kGy or less, and decomposition and denaturation of collagen occur under conditions of 200 kGy or more. In the manufacturing method according to the present invention, step 4) is a step of drying the filtered solution after stirring and according to the following specific method.

 i) obtaining a precipitate from the filtrate which filtered the stirring liquid;

 ii) obtaining a supernatant after dissolving the precipitate of step i) in an acid; iii) adding a salt to the supernatant of step ii) to obtain a precipitate; And iv) the precipitate of step iii) is dissolved in acid, diluted, and then lyophilized, but not always limited thereto. In addition, the present invention provides a method for producing atelo collagen comprising the step of treating the acid-soluble collagen prepared by the above method with a protease and drying.

 The protease is preferably pepsin or trypsin, more preferably pepsin, but is not limited thereto.

 The protease preferably adds 1 to 10 (w / w)%, more preferably 3 to 6 (w / w)%, and most preferably 5 (/)%, but is not limited thereto.

 The protease removes the telopeptide of the collagen (telo peptide), and since the helix structure in the terminal portion of the collagen molecule is removed, since the antigenicity is removed, it is easy to use as a biomolecule.

The drying is fast freezing at -178 to -70 ^° C, but is not limited thereto.

 The present invention provides a specific method for producing atelo collagen from jellyfish comprising the following steps.

 a) dissolving acid-soluble collagen in a mixture of acid and pepsin and then stirring;

 b) obtaining a precipitate from the agitated mixture of step a), dissolving in acid, and then adding salt to precipitate collagen; And

c) dissolving the precipitated collagen of step b) in acid, diluting and freezing. In the present invention, the method for separating collagen from jellyfish using radiation is to wash and pulverize the jellyfish, immerse it in an acid solution, irradiate acid-soluble collagen by irradiation, and then lyophilize by treating with pepsin. It can manufacture.

Specifically, as shown in the schematic diagrams of FIGS. 1 and 2, the jellyfish are washed and pulverized, the pulverized jellyfish is immersed in an acidic solution, the solution is irradiated with radiation, stirred, and the precipitate is filtered to obtain a precipitate from the filtrate. Next, after dissolving in acid, a supernatant was obtained, and the salt-added precipitate was dissolved in acid, diluted, and then lyophilized to extract acid-soluble collagen. Then, the acid-soluble collagen was dissolved in a mixed solution of acid and pepsin and stirred. After obtaining a precipitate from the agitated solution, it was dissolved in acid, and then salt was added to precipitate collagen, dissolved in acid, diluted, and then lyophilized. Atelo collagen can be prepared. In a specific embodiment of the present invention, Nomura pulverized jellyfish (Afe »(¾ e / nomuri Kishinouye) was washed with distilled water and then pulverized, soaked in acetic acid, irradiated with gamma rays, and stirred, and filtered with stirring. After dilution with filtrate, the precipitate was dissolved in acetic acid to obtain a supernatant. Then, sodium chloride was added to the supernatant to obtain a precipitate, which was then dissolved in acetic acid and lyophilized to obtain acid-soluble collagen. Acetic acid and pepsin were dissolved and stirred, and after obtaining a precipitate from the agitated solution, it was dissolved in acid, and then salt was added to precipitate collagen, dissolved in acid, diluted, and lyophilized to prepare atelo collagen (Fig. 1 and 2) to analyze the weight and particle size of the jellyfish according to the grinding in the experimental example of the present invention And, when pulverizing the seaweed before lyophilization, the weight is reduced by 25%, and as the pulverization time is continued, the particles of the lyophilized jellyfish become smaller, and especially when pulverized for 60 seconds, it has a particle size of about 128 JMII ( 3 and 4, and Table 1 and Table 2) As a result of analyzing the acid-soluble collagen extraction degree according to the extraction time in the experimental example of the present invention, the collagen yield when the collagen is extracted after stirring for 3 days or 5 days It was found to increase significantly (see FIG. 5).

 As a result of analyzing the acid-soluble collagen extraction degree according to the radiation dose in the experimental example of the present invention, the yield of collagen extracted as the radiation dose increases, especially collagen yield is significantly increased by irradiation dose of 10 kGy and 25 kGy It increased, and turned slightly yellow at 100 kGy (see FIG. 6).

 As a result of analyzing the atelocollagen extract according to the radiation dose in the experimental example of the present invention, it was confirmed that the weight change is less in the case of atelocollagen having a high radiation dose (see FIG. 7).

 As a result of analyzing the chemical and thermal properties of collagen according to the radiation dose in the experimental example of the present invention, by confirming that the collagen extracted by irradiation from the jellyfish according to the present invention has a spectral pattern similar to that of animal-derived collagen, radiation Did not affect the chemical and thermal properties (see Figs. 8 and 9).

 As a result of analyzing the components of atelo collagen according to the radiation dose in the experimental example of the present invention, it was confirmed that the components of the jellyfish-derived atelo collagen and animal-derived collagen prepared by the present invention were similarly irradiated with 10 kGy irradiation dose. (See Figure 10 and Table 3)

 Therefore, using the method for separating acid-soluble collagen from the jellyfish of the present invention and the method for producing atelo collagen, the yield of separating collagen is very high compared to the method of treating only conventional chemicals, and the use of chemicals can be reduced. Costs are reduced, and jellyfish, which are not good for the ecosystem, can be used to help prevent pollution. Hereinafter, the present invention will be described in detail by Examples and Experimental Examples.

 However, the following Examples and Experimental Examples are merely illustrative of the present invention, and the content of the present invention is not limited by the following Examples and Experimental Examples.

Example 1 Jellyfish Washing and Milling

The Nomura Inverted Jellyfish (Nemop i 1 ema nomur i Ki sh inouye) was obtained from the National Fisheries Research and Development Institute and served with ice in an icebox at Bihonghang (Gunsan, Korea). Transported refrigerated. After washing the salted state jellyfish to 4 ^° C and washed with distilled water, and replace the distilled water at 4 ^° C for 3 days. Then, the washed jellyfish was pulverized with a blender and then drained with a strainer. The dried jellyfish was then lyophilized and ground in a blender.

Example 2 Immersion of Jellyfish in Acid Solution and Irradiation

The jellyfish sliced in Example 1 was soaked in 0.5 M acetic acid (acetic acid, glacial grade, Merck (Darmstadt, Germany)), and then gamma rays ( ⁶⁰ Co source, Pencil type, MDS Nordion, Cananda) were 10 kGy / hr. Irradiated at a dose of 10 to 100 kGy and stirred at 4 ° C for 2 weeks.

Example 3 Extraction of Acid Soluble Collagen from Jellyfish

The stirred solution obtained in Example 2 was filtered, and then the filtrate was diluted with 0.02 M sodium hydrogen phosphate (N¾HP0 ₄ , Sigma (St. Luis MO, USA)) and l: 3 (v / v) for dialysis. The precipitate was then obtained using a centrifuge (2000 rpm, 6 min). This precipitate was dissolved in 0.5 M acetic acid again, and the supernatant was obtained again using a centrifuge (2000 rpm, 6 min). Then, sodium chloride (Sodium chloride, NaCl, Sigma) was added to the supernatant to obtain a precipitate, which was then dissolved in 0.5 M acetic acid, diluted to 0.1 M acetic acid, and lyophilized to obtain acid-soluble collagen.

Example 4 Preparation of Atelocollagen

The <Example 3> The acid-soluble collagen, separated in 0.5 M acetic acid and 5 w / w% pepsin (pepsin, EC 3.4.23.1, 2 x crystallized, Tokyo chemical industry, Japan) was dissolved in a solution heunhap 4 ^° C Stirred for 24 h. The stirred mixture was diluted with 0.02 M disodium hydrogen phosphate to obtain a precipitate by centrifugation, and then dissolved in 0.5 M acetic acid, followed by addition of sodium chloride to a concentration of 0.9 M to precipitate collagen. It was diluted so that then the dissolved collagen was again precipitated in 0.5 M acetic acid ^'0.1 M acetic acid and then freeze-dried collagen with ahtel Prepared.

Experimental Example 1 Jellyfish Weight Analysis According to Grinding

 Since the Nomura-infused jellyfish is a large jellyfish with more than 90% of its body water, it is necessary to reduce the volume before freeze-drying of the jellyfish. Therefore, the jellyfish washed by the method described in Example 1 was pulverized with a blender to check the weight of the jellyfish according to the grinding.

 As a result, as shown in Figure 3 and Table 1, it was confirmed that the weight of the jellyfish before and after crushing is reduced by about 25% (Fig. 3 and Table 1).

 Table 1

Experimental Example 2 Jellyfish Particle Size Analysis According to Grinding Time

 In order to analyze the jellyfish particle size according to the grinding time, the jellyfish lyophilized by the method described in <Example 1> was pulverized for 0, 15, 30, 45 and 60 seconds, and then the particle size of the jellyfish pulverized by electron microscope was confirmed. It was.

 As a result, as shown in Figure 4 and Table 2, the particle size of the jellyfish decreases with the grinding time, in particular, when the freeze-dried jellyfish is pulverized for 60 seconds, up to about 17 times the particles than when crushed for 15 seconds It was confirmed that the size was reduced (Figure 4 and Table 2).

 Table 2

<실험예 3>추출시간에 따른산가용성 콜라겐의 분석

Experimental Example 3 Analysis of Acid-Soluble Collagen According to Extraction Time

 In order to analyze the degree of acid-soluble collagen extraction according to the extraction time

The jellyfish sliced in Example 1 was soaked in 0.5 M acetic acid (acetic acid, glacial grade, Merck (Darmstadt, Germany)) and irradiated with gamma rays at 10 and 25 kGy and then stirred for 1, 3 and 5 days at 4 ^° C. Then, the collagen was extracted by the method of <Example 3>, and the weight of the obtained collagen was measured to determine the yield according to the following [Equation 1] (Fig. ₅₎ ᅳ

[Equation 11

丁^{¾ j} om ^ m

 As a result, as shown in Figure 5, when the collagen was extracted after stirring for 3 or 5 days after stirring for 1 day, the collagen yield was increased, the irradiation dose of 25 kGy than the irradiation dose of 10 kGy When the gamma ray was irradiated with it, it was confirmed that the collagen yield was further increased (FIG. 5). Experimental Example 4 Analysis of Acid-Soluble Collagen According to Radiation Dose

To analyze the degree of acid-soluble collagen extraction according to the radiation dose, the jellyfish sliced in Example 1 was soaked in 0.5 and 1 M acetic acid and irradiated with 0, 10, 25, 50, and 100 kGy, and then 4 ^° C. After stirring for 2 weeks at <3> collagen was extracted, and the weight of the obtained collagen was measured to determine the yield according to the above [Equation 1] (Fig. 6).

 As a result, as shown in Fig. 6, when the 0 kGy at 100%, the collagen yield extracted as the irradiation dose increased, and the collagen yield extracted up to 421,20 ± 67.66% at the irradiation dose of 25 kGy It was found to increase significantly (FIG. 6).

Experimental Example 5 Analysis of Atelocollagen According to Radiation Dose

In order to analyze the atelocollagen extract according to the radiation dose After dipping the jellyfish sliced in <Example 1> in 0.5 and 1 M acetic acid and irradiated with gamma rays 0, 10, 25, 50 and 100 kGy and stirred for 2 weeks at 4 ^° C. Acid-soluble collagen was isolated. Subsequently, the isolated acid-soluble collagen was immersed in a mixed solution of 0.5 M acetic acid and 5 w / w% pepsin (pepsin, EC 3.4.23.1, 2 x crystallized, Tokyo chemical industry, Japan) for 24 hours at 4 ° C. After stirring, the atelo collagen was extracted by the method of <Example ₄ >, and the weight was measured to determine the extraction rate according to the following [Equation 2].

 [2]

^ ^ _sw ¾ Cher: Lihua € Locoli · By _{y Λ m}

丁 ^^ ^" ᅳ 쳉 Availability Coke ¾ Muke 무

As a result, as shown in Figure 7, the weight of the atelo collagen produced by the pepsin is reduced, but the weight change is less in the case of the high dose of atelo collagen, the extraction amount when extracting collagen using radiation It was confirmed that can be increased (Fig. 7).

Experimental Example 6 Chemical Characterization of Collagen According to Radiation Dose

 The chemical properties of acid soluble collagen were analyzed using an ATR-FTIR spectrophotometer.

Specifically, ATR-FTIR spectrophotometer was used to extract rat tail type I collagen from collagen extracted from gamma rays 0 and 10 and 25 kGy and terrestrial animal-derived collagen as shown in Experimental Example 4. (Bruker TEMSOR 37, Bruker AXS. Inc., Germany). Strum measured the range of 500-4000 cnf ¹ , and ATR mode and head count were analyzed under 64 conditions and 4 cm- ¹ resolution. As a result, as shown in Figure 8, the chemical properties of the acid-soluble collagen was confirmed. Marine collagen has a spectral pattern similar to that of animal-derived collagen. The regions of amides A, I and II are directly related to the form of the polypeptide. Amide A region (3400-3440 cm— ¹ ) is NH The amide I region (160 cnf ¹ ) is associated with stretching vibrat ions of the carbonyl group of the peptide and is most useful for examining the secondary structure of proteins. It is used. The amide Π region (-1550 cm– ¹ ) is associated with NH bending and CN stretching, and with the triple helical structure of collagen. In the case of jellyfish collagen, peaks of amide I, amide II, and amide A were found at 1635 cm ^"1 , 1530 cm ^-1 , and 3280 cn ¹ (FIG. 8).

Experimental Example 2 Thermal Characteristics Analysis of Collagen According to Radiation Dose

 Thermal properties of acid-soluble collagen were analyzed using differential scanning calorimeters-!-,

 Specifically, the collagen and rat tail type 1 collagen extracted by gamma rays 0, 10, and 25 kGy irradiation to sliced jellyfish as described in <Experimental Example 4> (differential scanning calorimeters) (TA Q100, TA instruments, USA). The measurement of the sample was carried out at a temperature rising rate of 10 ° C / min in nitrogen.

Measured up to 0 ~ 300 ° C.

 As a result, as shown in Figure 9, both acid value collagen extracted after 10 kGy and 25 kGy irradiation according to the heat change shows a patternol similar to collagen extracted from jellyfish that do not irradiate gamma rays, especially acid value collagen extracted after 10 kGy irradiation By confirming that the pattern is almost the same as the collagen extracted from the jellyfish irradiated with gamma rays, it was confirmed that irradiation for obtaining a high concentration of collagen from jellyfish does not affect the thermal properties (Fig. 9).

Experimental Example 8 SDS-PAGE Analysis of Atelocollagen According to the Radiation Dose

 To analyze the components of atelocollagen according to the radiation dose, SDS-

PAGE was performed.

Specifically, electrophoresis was measured by the method of Lae '1K1970 using Mini-Protean 3 (Bio-Rad Laboratories, Hercules, CA). Polyacryllatnide gel is a stacking gel and a resolving gel. Each was used at a concentration of 5%. The concentration of the atelo collagen sample extracted from jellyfish irradiated with gamma rays at 0, 10 and 25 kGy after dipping in 0.5 M acetic acid as in <Experiment 5> was used to make 20 mg / m £ with distilled water, 10% SDS containing 0.25 M Tris-HCKpH 6.8), 20% glycero the (Glycerol), 5% 2-mercapto ethanol (2-Mercaptoethanol), then heunhap the bromophenol blue (bromophenol blue) of 0.1% 100 ^° Heated at C for 3 minutes. Samples were prepared in the same manner as the jellyfish collagen to compare and analyze the mouse tail type 1 collagen with the control protein. The prepared jellyfish collagen sample and rat tail type 1 collagen sample were injected into the polyacrylamide gel and subjected to periodic electrophoresis at 20 mA / gel. After electrophoresis, the gel was stained with 0.25¾ (w / v) Coomassie brilliant blue R250 and bleached with a mixture of methanol and acetic acid to make rat tail type 1 collagen and jellyfish collagen. Was analyzed comparatively.

 As a result, as shown in Figure 10, the mouse tail type 1 collagen consists of two αΐ- chain, α2 α chain, β-component (crosslinked dimer of α_chain), In the case of jellyfish-derived atelo collagen, α-chain and α2-chain showed the same mobility, but β-element was faint (FIG. 10).

Experimental Example 9 Amino Acid Analysis of Atelo Collagen According to the Radiation Dose

 As in <Experiment 5>, amino acid analysis of each of atelo collagen and rat tail type 1 collagen extracted from jellyfish irradiated with gamma rays at 0 and 10 kGy after immersion in 0.5 Μ acetic acid was performed.

 As a result, as shown in Table 3, both jellyfish collagen and rat tail type 1 collagen occupy a large amount of glycine, alanine and prol ine, thus characterizing typical collagen characteristics. Showed. In particular, it was confirmed that the amino acid components of atelocollagen irradiated with gamma-rays and atelocollagen irradiated with 10 kGy hardly changed. In addition, the jellyfish collagen of the present invention was confirmed that the content of hydroxyproline (hydroxyproline) is lower than that of mammalian rat tail type 1 collagen (Table 3).

Table 3

Claims

[Claims]  [Claim 1]  1) washing and pulverizing jellyfish;  2) dipping the pulverized jellyfish of step 1) in an acidic solution;  3) irradiating and stirring the solution of step 2); And  4) A method for separating acid-soluble collagen (collagen) from the jellyfish comprising the step of filtering and stirring the stirring of step 3). [Claim 2]  The method of claim 1, wherein step 1)  I) crushing the washed jellyfish;  II) lyophilizing the ground jellyfish of step I); And  III) A method for separating acid-soluble collagen from jellyfish, characterized in that the step of pulverizing the lyophilized jellyfish of step II). [Claim 3]  3. The method of claim 2, wherein the lyophilized jellyfish of step III) is ground to a particle size of 100 to 3000. [Claim 4]  The method for separating acid-soluble collagen from jellyfish according to claim 1, wherein the acidic solution of step 2) is any one selected from the group consisting of acetic acid, citric acid and formic acid. [Claim 5] The method of claim 1, wherein the acidic solution of step 2) is a method of separating acid-soluble collagen from jellyfish, characterized in that the concentration of 0.01 M to 2.0 M. [Claim 6]  The method of claim 1, wherein the radiation of step 3) is a gamma ray or an electron beam. [Claim 7]  The method of claim 1, wherein the radiation of step 3) is irradiated with an irradiation dose of 5 kGy to 200 kGy. [Claim 8]  The method of claim 1, wherein step 4)  i) obtaining a precipitate from the filtrate which filtered the stirring liquid;  v) obtaining a supernatant after dissolving the precipitate of step i) in an acid; iii) adding a salt to the supernatant of step ii) to obtain a precipitate; And iv) dissolving the precipitate of step iii) in an acid, diluting it, and then lyophilizing the acid-soluble collagen from the jellyfish. [Claim 9]  A method for producing atelo collagen comprising the step of treating acid-soluble collagen prepared by the method of any one of claims 1 to 8 with a protease and then drying. [Claim 10]  10. The method of claim 9, wherein the protease is pepsin or trypsin. [Claim 11] 10. The method according to claim 9, wherein the protease is characterized by adding 1-10 (\%). [Claim 12]  The method of claim 9, wherein the protease removes telopeptides of collagen. [Claim 13] The method of claim 9, wherein the drying is rapid freezing at -Γ78 to -70 ^° C. [Claim 14]  The method of claim 9,  a) dissolving the acid-soluble collagen in a mixed solution of acid and pepsin and then stirring;  b) obtaining a precipitate from the agitated mixture of step a), dissolving it in acid, and then adding salt to precipitate collagen; And  c) a method for producing atelo collagen, characterized in that the step of dissolving the precipitated collagen in step b) and diluting, followed by freezing.