JP2007051191A - Method for recovering collagen from jellyfish - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To recover collagen, which is a useful material, from jellyfish by solubilizing the collagen efficiently in a native state. <P>SOLUTION: The method for recovering collagen comprises the steps of: freezing jellyfish, thawing the frozen jellyfish to activate an endogenous enzyme of jellyfish itself to start the decomposition reaction of jellyfish, mixing the thawed jellyfish to solubilize the collagen of jellyfish in a native state to form a neutral salt solution containing native collagen, and recovering the native collagen from the neutral salt solution. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a method for efficiently solubilizing and recovering collagen, which is a useful substance from jellyfish, in an undenatured state.

  A variety of species that are not subject to fishing are distributed in the ocean, and they serve to protect the marine ecosystem, and sometimes anomalies of a single species have a significant impact on fisheries and other industries. May give. In recent years, large amounts of moon jellyfish and Echizen jellyfish have occurred in various places along the coast of Japan, causing a great deal of damage to the fishery. In addition, a large amount of various marine organisms gather in the cooling water intakes of factories and power plants, including not only fishery species but also non-fishery species such as moon jellyfish and Echizen jellyfish. Not only are these precious marine biological resources not used effectively, but they also require significant costs for their removal and processing.

  Therefore, it has been considered to effectively use jellyfishes such as moon jellyfish and Echizen jellyfish that gather at the water intake. As a method for effectively utilizing jellyfishes, it has been proposed to extract collagen from jellyfish. Collagen produced mainly from livestock animals is in great demand for food, pharmaceuticals and industrial products. Since the occurrence of BSE, there has been a problem in terms of raw material supply and safety against relying solely on livestock animals for collagen raw materials. Accordingly, various attempts have been made to utilize unused marine animals such as jellyfish as collagen raw materials.

  As a technique for extracting collagen from jellyfish, for example, there is a method of freezing and thawing moon jellyfish collected from the sea at −20 ° C. to obtain about 4% solids, separating water-soluble protein from this and using it as collagen. It has been devised (see, for example, Patent Document 1).

  In addition, the jellyfish is shredded to a size of 10 mm square or less and extracted by solubilizing it in a buffer solution of pH 6.0 to 8.0, and the collagen extracted in the buffer solution is precipitated by a salting-out device. Then, after dehydrating the precipitated collagen, a method of freezing or drying has been devised (for example, see Patent Document 2).

In addition, a first step of crushing / chopping jellyfish, a second step of decomposing / solubilizing the crushed / chopped jellyfish, and a third step of roughly refining useful substances from the decomposed / solubilized jellyfish Thus, a method of efficiently roughly extracting jellyfish-derived protease and collagen has been devised (see, for example, Patent Document 3).
JP 2003-321497 A JP 2004-99513 A JP 2001-178492 A

  However, the properties of the water-soluble protein obtained by the method described in Patent Document 1 are not described in the document, and it is completely unknown what purity the collagen has. Since native collagen is inherently insoluble in water, it is presumed that the recovered collagen has undergone denaturation or molecular weight reduction. When using collagen recovered by such a method and subjected to denaturation or low molecular weight, it is difficult to select an application.

  Further, in the method of Patent Document 2, extraction is performed by adding about 5 volumes of buffer solution to 1 volume of moon jellyfish, so that a large space and buffer solution for preparing buffer solution and collagen extraction are prepared. Cost is required.

  In the method of Patent Document 3, it is considered that the treatment temperature is preferably 27 ° C. to 37 ° C. in the second step, and it is highly possible that collagen is denatured in this temperature range. Therefore, it is assumed that when the enzyme treatment is performed under these conditions, the collagen is remarkably reduced in molecular weight, but the properties of the recovered collagen are not described in the literature, and what purity is the collagen? Is completely unknown. Furthermore, since about 5 volumes of water is added to 1 volume of moon jellyfish during extraction, there remains a problem that a large space for extraction is required.

  The present invention has been made to solve such a conventional problem, and aims to provide a method for efficiently solubilizing and recovering collagen, which is a useful substance from jellyfish, in a native state. It is.

  The invention according to claim 1 of the present invention includes a freezing step for freezing jellyfishes, and a thawing for thawing frozen jellyfishes in order to activate endogenous enzymes possessed by the jellyfishes themselves and initiate a jellyfish decomposition reaction. A step of stirring the thawed jellyfish in order to solubilize the collagen possessed by the jellyfish in a native state to produce a neutral salt solution containing the native collagen; A recovery step of recovering native collagen.

  In addition, the invention according to claim 2 of the present invention has a freezing step in which jellyfish are shredded and frozen, and an endogenous enzyme possessed by the jellyfish itself is activated to initiate a decomposition reaction of the jellyfish. Thawing the frozen jellyfish to recover solubilized collagen from the neutral salt solution to solubilize the collagen in its native state to produce a neutral salt solution containing the native collagen And a recovery step.

  According to the method for producing collagen from jellyfishes of the present invention, it is possible to recover native collagen by a simple operation by effectively utilizing an endogenous enzyme possessed by the jellyfish itself.

  Hereinafter, a method for recovering collagen from jellyfish according to an embodiment of the present invention will be described in detail with reference to the drawings.

  FIG. 1 is a diagram for explaining each step of a method for recovering collagen from jellyfish according to an embodiment of the present invention.

(Freezing process)
In step 101 (in the figure, step is abbreviated as S. The same shall apply hereinafter), the jellyfish is stored as it is or after being shredded at a temperature at which the jellyfish freezes or at a temperature lower than that.

  Because jellyfish often occur at irregular times, it is necessary to preserve the raw materials for a long period of time. However, jellyfish have a high water content in the body, and the progress of decay is particularly rapid under high temperature conditions. Therefore, by freezing and storing jellyfish, a large amount of raw jellyfish can be stored easily and efficiently and used as a resource.

  Here, the temperature at which the jellyfish is cryopreserved is not particularly limited as long as it is below the temperature at which the jellyfish can be frozen (around −2 ° C.), but it is preferably cryopreserved at −20 to −30 ° C.

(Thawing process)
Step 102 is a step of thawing frozen jellyfish at a predetermined temperature at which the collagen contained in the jellyfish can maintain an undenatured state. This thawing step is a step of stirring, which will be described later, in which jellyfish is thawed to a fluid and stirrable state to activate endogenous enzymes possessed by jellyfish themselves and start a decomposition reaction.

  As a result of intensive studies, the present inventors have found that an enzyme that solubilizes collagen works when jellyfish is thawed.

  This is because the electrophoretic pattern of the solubilized collagen obtained by this embodiment is reproducible, and is very similar to the electrophoretic pattern of collagen decomposed by matrix metalloprotease, and an inhibitor of metalloprotease This is because it has been clarified that the cause of solubilization is due to enzymatic degradation from the fact that the addition of (EDTA) suppresses solubilization of collagen.

  In addition, as a result of intensive studies, the present inventors have found that destruction of tissues in jellyfish by freezing and thawing helps solubilize collagen.

  In general, in the case of slow freezing (freezing method in which the temperature is slowly lowered), the ice crystals formed in the tissue become larger and the tissue is destroyed by the ice crystals. This is because the enzyme is leaked to the outside of the cell, the localized enzyme is dispersed, and the chance of contact between the protein and the enzyme in the tissue is increased, so that the enzymatic degradation is promoted.

  Further, as a result of intensive studies, the present inventors have found that the denaturation temperature of collagen possessed by jellyfish is around 30 to 34 ° C., and the denaturation starts gradually from the latter half of the 20 ° C. range.

  FIG. 7 is a graph showing the denaturation temperature of collagen obtained by the example of the present invention. FIG. 7A shows the denaturation temperature of collagen under acidic conditions, and FIG. 7B shows the denaturation temperature of collagen under neutral conditions.

  As shown in the figure, not all the collagen molecules tested were denatured at that temperature (33.7 ° C under neutral conditions) at once, but gradually began to denature at around 27-30 ° C. It will follow the Yamagata process in which the number of unmodified molecules decreases after reaching the peak and then reaching the upper half of the 30 ° C range.

  From the figure, it can be seen that the temperature at which no denaturation is considered to occur may change slightly due to subtle differences in conditions, so that it is sufficient to keep the temperature to 25 ° C. or less on the safe side.

  Therefore, in this thawing step, jellyfish are thawed at the upper limit of thawing temperature of the order of 20 ° C., preferably at a temperature of 25 ° C. or lower where it has been confirmed that no denaturation of collagen occurs. This makes it possible to maintain the undenatured state of collagen.

  Moreover, the lower limit temperature for thawing may be a temperature at which frozen jellyfish can be thawed (around −2 ° C.), but as a result of the evaluation by the present inventors, at a temperature of 4 ° C., the jellyfish itself Since it has been confirmed that the endogenous enzyme possessed has activity, thawing should be performed at a temperature of 4 ° C. or higher, which is the temperature at which the endogenous enzyme possessed by the jellyfish itself is active.

  More preferably, in this thawing step, thawing is performed in a temperature range of 4 ° C. to 10 ° C.

(Stirring process)
In step 103, the thawed jellyfish is agitated to solubilize collagen in an undenatured state. By this stirring process, the body tissue of jellyfish undergoes degradation by endogenous enzymes, almost completely disintegrates and becomes liquid. The body fluid of jellyfish contains salt (about 0.5 mol / l) comparable to seawater, and pH is 7.4-7.9. For this reason, when the body tissue of jellyfish is liquefied, it becomes a neutral salt solution containing unmodified collagen without adding a buffer solution or the like.

  The stirring method may be a general method such as inserting a propeller or the like into a container containing jellyfish and rotating it. Moreover, it is preferable that it is the stirring method which does not produce a bubble at the time of stirring.

  In the stirring step, stirring is performed until the jellyfish are in a solution state or until they are in a steady state (a state where dust and tissue pieces are still left but do not change any more). When there is insoluble matter (dust, tissue fragments, and other contaminants) that remains undissolved during the stirring step, a step of removing them by filtration or centrifugation may be provided separately.

  In addition, the temperature during the stirring step is preferably within the temperature range where collagen denaturation does not occur, and within the temperature range where the endogenous enzyme of the jellyfish itself is active, as in the thawing step. . More preferably, in this stirring step, stirring is performed in a temperature range from 4 ° C to 10 ° C.

  In addition, when shredded before freezing jellyfish, depending on the size of the tissue piece after shredding, it becomes a neutral salt solution just by thawing without performing this stirring step. The process can be omitted.

(Recovery process)
In step 104, solubilized native collagen is recovered by salting out. The salting-out method is suitable for collecting undenatured collagen because only undenatured collagen is collected and can be used for simple purification.

  Undenatured collagen is completely precipitated by sodium chloride with a final concentration of 4.4 mol / l. Therefore, sodium chloride is added to the obtained neutral salt solution to a concentration of 4.4 mol / l and stirred efficiently. Native collagen can be recovered.

  The salt concentration is not limited to the above-mentioned concentration, and sodium chloride is added so as to obtain a salt concentration at which the undenatured collagen is completely precipitated, depending on the jellyfish that collects the collagen. Regarding the salt to be used, not only sodium chloride but also a salt that can perform the same function (for example, ammonium sulfate, potassium chloride, sodium sulfate, etc.) is applicable.

  In the present embodiment, collagen is recovered by salting out, but the recovery method is not limited to this, and for example, an ultrafiltration method or a chromatography method can be used. Further, once the neutral salt solution is concentrated by the ultrafiltration method and then salted out, salt consumption can be suppressed.

  Moreover, in this collection | recovery process, it is preferable to carry out the temperature in a process within the temperature range which denaturation of collagen does not occur. More preferably, in this recovery step, the recovery is performed in a temperature range from 4 ° C to 10 ° C.

  As described above, according to the method for recovering collagen from jellyfish of the present embodiment, the jellyfish can be frozen and thawed at a temperature at which the jellyfish freezes or after being shredded. After thawing at any temperature from 25 ° C. to 25 ° C., the collagen can be solubilized in an undenatured state by stirring and recovered by salting out.

  According to the method for recovering collagen from jellyfish according to the present embodiment, no space other than the space occupied by the jellyfish itself is required, and a reagent necessary for preparing a buffer solution is not required. By stirring after thawing, the jellyfish body tissue undergoes degradation by endogenous enzymes, almost completely disintegrates and becomes liquid. When the body tissue of jellyfish is liquefied, it becomes a neutral salt solution containing undenatured collagen.

  By adding sodium chloride to the obtained neutral salt solution at a concentration of 4.4 mol / l and stirring, the native collagen can be recovered efficiently and with a simple operation. Furthermore, by freezing and thawing jellyfish under neutral conditions, degradation occurs with good reproducibility, and by performing salt precipitation under neutral conditions, collagen exhibiting stable properties can be obtained.

  Regarding the mechanism of tissue disintegration due to agitation after thawing, the properties of the obtained collagen were examined by biochemical techniques, and as a result, it was mainly degraded by matrix metalloproteases (metal-requiring collagenolytic enzymes). Conceivable. Further, the collagen obtained by the method for recovering collagen from jellyfish according to the embodiment of the present invention was confirmed to be a native collagen having a triple helical structure retained by precipitation with respect to sodium chloride or differential scanning calorimetry. .

  Examples of the method for recovering collagen from jellyfish of the present invention will be described below. However, the method for recovering collagen from the jellyfish of the present invention is not limited to the following examples.

(Freezing / thawing / stirring method of jellyfish tissue)
An outline of the jellyfish tissue freezing and thawing method employed in the evaluation of this example is shown in FIG.
2A is a diagram showing a freezing / thawing method A, and FIG. 2B is a diagram showing a freezing / thawing method B.

  In the freezing / thawing method A, the tissue was shredded by homogenizing at 5,000 rpm for about 2 minutes in advance using a homogenizer before freezing. 500 g of a chopped sample was placed in a 1 liter square jar, frozen at -30 ° C for about 2 weeks, and thawed at 4 ° C for 24 hours.

  On the other hand, in the freezing / thawing method B, 3 kg of moon jellyfish was placed in a plastic container in a hole, frozen at -30 ° C for about 2 weeks, and then thawed at 4 ° C for 24 hours. At that time (half-thawed state), the mixture was homogenized using a homogenizer in the same manner as described above, and stirred at 4 ° C. until it became a solution (about 2 hours).

  The collagen solution obtained by the freezing / thawing methods A and B was centrifuged at 10,000 g × 20 minutes to remove the remaining undegraded suspension.

(Collagen recovery method)
In order to collect collagen solubilized by an endogenous enzyme, two methods shown in FIG. 3 were employed in the evaluation of this example. FIG. 3 (a) is a diagram showing precipitation under neutral conditions (referred to as precipitation method I), and FIG. 3 (b) is a diagram illustrating precipitation under acidic conditions (referred to as precipitation method II). .

  In precipitation method I (precipitation under neutral conditions), sodium chloride was added to the obtained collagen solution to a final concentration of 4.4 M, and the precipitated collagen was collected by centrifugation. In addition, since the pH of the obtained collagen solution was in the range of about 7.4 to 7.8, in this method, collagen is precipitated under neutral conditions. The obtained precipitate was washed with 0.5 M acetic acid containing 2 M sodium chloride, dialyzed against distilled water, and lyophilized.

  In precipitation method II (precipitation under acidic conditions), acetic acid is added to the obtained collagen solution to a final concentration of 0.5M, pH is adjusted to 2.2, and then sodium chloride is added to a final concentration of 2M. To precipitate collagen. The resulting precipitate was collected by centrifugation, dialyzed against distilled water, and lyophilized.

  As shown in FIG. 4, the collagens prepared this time by the combination of freezing / thawing methods A and B and precipitation methods I and II were respectively collagen A (combination of freezing / thawing method A and precipitation method I. “Example 1 ), Collagen B (combination of freezing / thawing method B and precipitation method I. “Example 2”), collagen C (combination of freezing / thawing method A and precipitation method II). Example 1 ”) and collagen D (combination of freezing / thawing method B and precipitation method II.“ Comparative example 2 ”).

(SDS-PAGE analysis of recovered collagen)
The lyophilized product or the fraction obtained by chromatography was subjected to SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) to examine their properties.

  FIG. 5 is a diagram showing SDS-PAGE patterns of collagens A, B, C and D. FIG. In the figure, “M” indicates a marker, “A” indicates collagen A, “B” indicates collagen B, “C” indicates collagen C, “D” indicates collagen D, and “E”. Indicates pepsin solubilized collagen (PSC) of moon jellyfish.

  Collagen A (Example 1) shows two major bands around 130-150K, two bands around 97K, and one band around 50K, etc. Showed a typical pattern exhibited by degraded collagen.

  Collagen B (Example 2) showed almost the same pattern as collagen A. Therefore, when precipitation method I (precipitation under neutral conditions) was used, the effect of freezing / thawing method on the properties of collagen was small. It is considered a thing.

  Collagen C (Comparative Example 1) basically showed a pattern similar to collagen A or B, but the band pattern around 97K was different.

  Collagen D (Comparative Example 2) was significantly different from other collagens in that it showed three bands near 130-150K and a large number of minor bands in the low-molecular region from 97K. . A possible cause of the significant decrease in the molecular weight of collagen D in collagen D is that the solubilized collagen may be side-degraded by acidic protease during salting out.

  Furthermore, the collagen recovered using precipitation method I showed a reproducible SDS-PAGE pattern, but when using precipitation method II, some differences were observed for each experiment.

  From the above results, it is considered that precipitation method I (salting out with 4.4 M sodium chloride under neutral conditions) is suitable for the recovery of collagen.

  When the precipitation method I was used, there was almost no difference depending on the freezing / thawing method. Therefore, the freezing / thawing method A (Example 1) can be used when the amount of jellyfish treatment is small. When a large amount of jellyfish needs to be processed, it is considered preferable to use the freezing / thawing method B (Example 2).

  Therefore, the validity of the recovery method of the present invention combining the freezing / thawing method A or B and the precipitation method I was proved.

(Properties of partially degraded collagen)
As described above, since collagen having a reproducible SDS-PAGE pattern was recovered when it was precipitated by precipitation method I, amino acid analysis was performed on collagen A (Example 1). It was compared with the amino acid composition of PSCs of PSC and Stomolophus nomurai (FIG. 6).

  As described above, since collagen A (Example 1) and collagen B (Example 2) exhibit substantially the same properties, the characteristics of collagen A described later may be considered as the characteristics of collagen B as they are.

(Amino acid analysis)
The amino acid analysis was performed by the following method. A 100 μg collagen A freeze-dried sample was dissolved in 100 μl 0.1 M hydrochloric acid to prepare a sample. The analysis was performed by the PICO TAG method using an amino acid analysis system (Waters PICO TAG system; Waters, Milford, Mass) equipped with an ODS column (Cosmosil 5C18-AR; 4.6 × 250 mm; Nacalai Tesque, Kyoto, Japan). The sample solution placed in the test tube was subjected to gas phase hydrolysis with 6M hydrochloric acid at 150 ° C. for 1 hour in a vial under reduced pressure. The hydrolyzate was neutralized with a methanol-ultra pure water-triethylamine (2: 2: 1) mixture. Phenylthiocarbamoyl derivative formed by the reaction of PITC and amino acids by adding a mixed solution of methanol-ultra pure water-triethylamine-phenyl isothiocyanate (PITC) (7: 1: 1: 1) to the neutralized hydrolyzate Were separated and quantified by the above amino acid analysis system.

  As shown in FIG. 6, collagen A generally showed an amino acid composition similar to moon jellyfish PSC and Echizen jellyfish PSC, but glycine (Gly) was as low as 260.7 residues per 1000 residues, and lysine (Lys) was 58.3. Because of its high residue, collagen A is considered to contain proteinaceous impurities.

  However, it can be said that the collagen sample obtained by only a simple operation of adding a salt to the solution obtained by freezing and thawing has a relatively high purity. Therefore, the effectiveness of the recovery method of the present invention was proved also from this point.

(Denaturation temperature)
Next, the denaturation temperature of the collagen obtained in this example was measured. Sample preparation for measurement of denaturation temperature was performed at 4 ° C., and measurement was performed under the following two conditions.

  (1) Acidic conditions: Collagen A (Example 1) A lyophilized sample and a jellyfish pepsin-solubilized collagen (PSC) lyophilized sample were dissolved in 0.1 M acetic acid to a concentration of 1 mg / ml. Note that the moon jellyfish PSC was not sufficiently solubilized, so the supernatant obtained by crushing with Polytron and further stirring overnight, followed by centrifugation (18,000 × g, 4 ° C., 15 minutes) was used as a measurement sample. .

  (2) Neutral conditions: Collagen A and PSC were suspended in 20 mM sodium phosphate buffer (pH 7.2) to a concentration of 1 mg / ml and homogenized with polytron as a measurement sample. .

  Samples were subjected to a differential scanning calorimeter (Micro DSC III, Setaram, France), 0-25 minutes (0 ° C), 25-165 minutes (0-70 ° C, 0.5 ° C / min), 165-182 minutes (70- (20 ° C, 3 ° C / min).

  FIG. 7 is a diagram showing the denaturation temperature of collagen A obtained in this example. FIG. 7A shows the denaturation temperature of collagen A under acidic conditions, and FIG. 7B shows the denaturation temperature of collagen A under neutral conditions.

  As a result, collagen A showed a relatively high denaturation temperature as a marine animal at 30.3 ° C under acidic conditions and 33.7 ° C under neutral conditions, and as a control, the common casbe skin ASC (28.8 ° C) and rainbow trout muscle measured under acidic conditions. It was 1.5 ° C and 5.4 ° C higher than ASC (24.9 ° C), respectively.

  The denaturation temperatures of other jellyfish collagens have been reported to be 27 ° C for Stomolophus nomurai, 26.0 ° C for Stomolophus meleagris, and 28.8 ° C for Rhopilema asamushi. All of these are measured using the viscosity change of the PSC acidic solution as the treatment temperature rises as an index, so they cannot be compared in general, but even though they are partially decomposed by moon jellyfish, they are comparable. Or the point which shows the modification | denaturation temperature beyond them is notable.

  PSC could not be measured because it did not dissolve sufficiently under acidic conditions, but it was measured at 34.0 ° C in neutral. Although the denaturation temperature is slightly lower than PSC, probably due to partial degradation, it shows a denaturation temperature higher than that of common kasbe or rainbow trout ASC. These results indicate that collagen A (Example 1) Is excellent in terms of thermal stability.

(Search for endogenous protease)
Finally, an inhibitor was used to search for an endogenous protease that acts upon tissue thawing. Thawed frozen samples of various serine, cysteine, and aspartate proteases and inhibitors of metalloproteases, and the amount of protein in the dissolved fraction was measured. Inhibitors of metalloproteases (EDTA) A significant decrease in the amount of protein was observed only in the experimental group to which was added.

  These results indicate that the main enzyme involved in solubilization of the jellyfish collagen is a metalloprotease (a metalloprotease, a protease that requires a metal for activity expression). Yes. In addition, considering this together with the results of SDS-PAGE, this enzyme is a matrix metalloprotease (a metal-requiring collagenolytic enzyme, which is a type of metalloprotease that cleaves the triple helical region of collagen. ). In addition, even when inhibitors for various serine-based, cysteine-based, and aspartic acid-based proteases were added, a decrease in the amount of protein was observed, so the possibility that these proteases are involved in solubilization of collagen cannot be denied.

It is a figure explaining each process of the collagen collection | recovery method from the jellyfish by embodiment of this invention. It is a figure which shows the outline | summary of the freeze / thaw method of the jellyfish structure | tissue employ | adopted for evaluation of a present Example. It is a figure which shows the outline | summary of the precipitation method employ | adopted for evaluation of a present Example. It is a figure which shows the combination of freezing / thawing method A, B and precipitation method I, II which were employ | adopted for evaluation of a present Example. It is a figure which shows the SDS-PAGE pattern of collagen A, B, C, and D. It is the figure which compared the amino acid composition of collagen A, PSC of moon jellyfish, and PSC of Echizen jellyfish. It is a figure which shows the denaturation temperature in the acidic condition of the collagen A obtained by the Example of this invention. It is a figure which shows the denaturation temperature in the neutral conditions of the collagen A obtained by the Example of this invention.

Claims (8)

A freezing step of freezing jellyfish,
A thawing step of thawing the frozen jellyfish in order to activate an endogenous enzyme possessed by the jellyfish itself and initiate a decomposition reaction of the jellyfish,
A stirring step of stirring the thawed jellyfish in order to solubilize the collagen possessed by jellyfish in a native state to produce a neutral salt solution containing the native collagen;
A recovery step of recovering the native collagen from the neutral salt solution;
A method for recovering collagen from jellyfish, comprising: Freezing process to shred and freeze jellyfish,
The jellyfish itself activates an endogenous enzyme to initiate a jellyfish decomposition reaction, solubilizes the collagen that the jellyfish has in its native state, and generates a neutral salt solution containing the native collagen. In order to thaw the frozen jellyfish,
A recovery step of recovering the native collagen from the neutral salt solution;
A method for recovering collagen from jellyfish, comprising: Each of the steps excluding the freezing step,
The method for recovering collagen from jellyfishes according to claim 1 or 2, wherein the jellyfish has a collagen at or below a predetermined temperature at which the collagen is maintained in an undenatured state.   The method for recovering collagen from jellyfish according to claim 3, wherein the predetermined temperature at which the collagen of the jellyfish can maintain an undenatured state is 25 ° C. Each of the steps excluding the freezing step and the collecting step is
The method for recovering collagen from jellyfish according to any one of claims 1 to 4, wherein the enzyme is carried out at a predetermined temperature or more at which an endogenous enzyme possessed by the jellyfish itself is active.   The method for recovering collagen from jellyfish according to claim 5, wherein the predetermined temperature at which the endogenous enzyme of the jellyfish itself is active is 4 ° C.   The method for recovering collagen from jellyfish according to any one of claims 1 to 6, wherein in the recovery step, the undenatured collagen is recovered by a salting-out method. The method for recovering collagen from jellyfish according to any one of claims 1 to 7, wherein the endogenous enzyme includes a metalloprotease.

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