JP2001178492A - Method of extracting useful substance from aquatic life - Google Patents

Abstract

PROBLEM TO BE SOLVED: To attain rough extraction of the useful substances in jellyfish efficiently. SOLUTION: This crude extraction process for useful substances from jellyfishes comprises the first step in which the jellyfishes are crushed and shred into pieces, the second step in which the shred jellyfishes are decomposed and solubilized, and the third step in which the useful substance from the jellyfishes are roughly purified whereby the protease or collagen originating from the jellyfishes can be efficiently extracted.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a method for extracting useful substances from aquatic organisms such as jellyfish.

[0002]

2. Description of the Related Art Many factories and power plants in the seaside use seawater as cooling water. However, especially in summer, a large amount of water organisms such as jellyfish and sea slugs floating in the sea are forced to reduce water intake.

[0003] The water intake is covered with a fence or the like in order to prevent ingestion of foreign matter or marine life such as jellyfish.
However, a large amount of jellyfish floating in the sea gets on the flow of water and closes the fence, and as a result the water level near the water intake drops, so the water intake must be reduced, and as a result, the water intake is reduced And of course, the efficiency of operations and power generation must be reduced.

[0004] Therefore, in order to maintain the operation and power generation efficiency, it is necessary to capture and remove jellyfish approaching the water intake to secure a sufficient water intake. Most jellyfish that are caught and removed near the water intake are generally jellyfishes, and because they are different from edible jellyfish and Echizen jellyfish and their closely related species, they are generally discarded as they are. The fact is that no proper reuse has been attempted.

[0005] On the other hand, Japanese Patent Application Laid-Open No. 6-217737 discloses an effective use of jellyfish as a raw material. This is because the jellyfish is pulverized into liquefied or fine pieces, and then pH (4.0 to 8.5) and temperature adjustment (30 to 3).
5 ° C.), then add a precipitant or coagulant to precipitate (aggregate) the protein, dehydrate it, and then heat dry it to try to use jellyfish effectively.

[0006] However, in such a technique, only jellyfish is pulverized or crushed, and only a process of precipitating a useful substance → recovery of a precipitate is performed. Since the protein is not decomposed and solubilized, many unreacted unreacted parts of the body components are included in the treatment system. Such an unreacted portion has a problem of lowering the efficiency of extracting useful substances. Furthermore, the protein recovered as it is can be used as feed or the like, but it is difficult to say that it is necessarily an effective technique for obtaining a useful substance from jellyfish.

[0007] Under these circumstances, there is a need for the development of a technology for extracting valuable substances with high added value that aims at efficiently treating jellyfish to be trapped and removed and for effective reuse.

[0008]

In the conventional method for recovering precipitates from jellyfish, since the protein has not been decomposed and solubilized, many unreacted portions remain unsolubilized, and the extraction efficiency of useful substances is increased. In addition, the recovered material was only a protein for a sample, and only useful substances with low added value could be extracted.

The present invention has been made in view of the above problems, and aims to improve the efficiency of extracting useful substances by reducing the unreacted portion, and to recover water-derived valuable added valuable substances derived from aquatic organisms. An object of the present invention is to provide a method for extracting useful substances.

[0010]

In order to solve the above-mentioned problems, the method for extracting useful substances of aquatic organisms according to the first aspect of the present invention is a method of extracting enzymes from the aquatic organisms by fragmenting and shredding the aquatic organisms. And a second step of causing the enzyme to act on the protein in the fragment to decompose the fragment into a liquid substance, and a third step of extracting a useful substance from the liquid substance. Features.

The gist of the present invention is to use enzymes in the body of aquatic organisms to degrade proteins of the aquatic organisms themselves. Therefore, in the state where the in-vivo enzyme is mixed with the protein, the conditions under which the in-vivo enzyme is activated, for example, temperature, pH,
The protein is decomposed by setting the ionic strength and the like to a predetermined enzyme activity enabling condition.

The term "aquatic organism" as used herein means a cnidarian other than seawater jellyfish or freshwater jellyfish, such as sea anemones and corals, shellfish such as shellfish, sea urchins, and mollusks such as sea urchins.
Similarly, useful substances can be extracted by the treatment method of the present invention from crustaceans such as octopus and squid, and crustaceans such as shrimp and crab.

[0013] The method for extracting useful substances of aquatic organisms according to claim 2 is characterized in that in claim 1, the enzyme is an alkaline protease. As this enzyme, it is desirable to use an enzyme present in the body of aquatic organisms, but it may be added from the outside in the second step. Especially when the enzyme content in aquatic cells is decreasing due to seasonal fluctuations,
The efficiency of the treatment can be improved by adding the enzyme extracted in advance.

[0014] The method for extracting useful substances of aquatic organisms according to claim 3 is characterized in that, in claim 1, the useful substance is collagen.

[0015]

DESCRIPTION OF THE PREFERRED EMBODIMENTS The above-mentioned object is achieved by the present invention described below.

The present inventors have searched for jellyfish, particularly useful substances contained in aquatic organisms such as water jellyfish which are removed and landed in water intakes for power plants and factories. It was found to have a large amount.

Moon jellyfish contains a large amount of collagen. Naturally soluble collagen, insoluble collagen, and bovine / pork bone and skin, which are actually used at present, are decomposed with acids, alkalis, enzymes and the like. The solubilized collagen or the like has a collagen subunit structure (α1, α1, α2) composed of two heterologous polypeptides, and
Collagen derived from jellyfish has a helical structure composed of three heterologous polypeptide chains (subunit: α1 · α2
・ Has α3) and collagen contains 3 sugars
It has a characteristic structure of being bound by 特 徴 5% by weight or more, and has an excellent skin water-retaining effect because it has skin occlusive ability while maintaining appropriate moisture retention and moisture release properties. In the present invention, such high-quality collagen can be recovered and extracted by decomposing proteins.

The moon jellyfish generally comprises 95 to 96% of water in the body components, but the other components are generally 1.7% protein and 0.9 carbohydrate.
%, Fat 0.0%, and ash content 2.1%, and most of them are proteins except water. For this reason, by efficiently decomposing this protein portion, it is possible to easily decompose moon jellyfish.

Therefore, the present inventors have made effective use of the above-mentioned alkaline protease contained in the jellyfish body to efficiently decompose and solubilize the jellyfish, thereby contributing to the structure of the body of the jellyfish jellyfish. They have found that it is possible to extremely easily extract useful substances such as collagen as a component, and have completed the present invention.

In the method of the present invention, the jellyfish is first crushed and shredded as a first step, so that the alkaline protease localized in the body of the jellyfish can be released into and out of the cell. Next, as a second step following the first step, the jellyfish body components are treated in an extremely short time by treating the crushed and shredded jellyfish under conditions in which the jellyfish-derived alkaline protease suitably acts. It can be easily decomposed and solubilized. That is, by the action of the protease released from the jellyfish in the first step, the protein involved in maintaining the structure of the jellyfish is hydrolyzed to decompose and solubilize the jellyfish. Further, following the second step, as a third step, crude substances such as alkaline protease and collagen are roughly extracted from the liquid obtained by decomposing and solubilizing the moon jellyfish. It is characterized by comprising the above three steps.

As a method of crushing and shredding water jellyfish in the first step of the present invention, any method can be employed as long as it can efficiently crush and shred water jellyfish, for example, a biaxial crusher. And a shredder device can be used.

In the second step of the present invention, the jellyfish crushed and shredded in the first step, or the crushed and shredded jellyfish and an aqueous medium are mixed to allow the jellyfish-derived protease to act efficiently. It decomposes and solubilizes jellyfish. In this step, the jellyfish is decomposed and solubilized within several hours to half a day by controlling the temperature, pH, and jellyfish load in the decomposition / solubilization treatment within a range in which the jellyfish-derived protease treatment can be suitably performed. It becomes possible. Further, by controlling the decomposition / solubilization treatment conditions as described above, the activity of the alkaline protease in the solubilization treatment solution after the treatment is maintained at a high level, and the collagen in the treatment solution is further reduced. Without any loss, it is possible to send each solubilized state to the crude extraction step, which is the third step. For this reason, it is possible to perform the rough extraction processing of the third step without performing complicated pretreatment at the subsequent stage of the second step.

Here, the solubilization in the present invention refers to a state in which it is almost completely dissolved in a solvent, and does not refer to a state in which the fine flakes are in a liquid state only by diffusion. Absent.

As the processing conditions in the second step, first, as the temperature conditions, conditions that do not affect the intended useful substance can be set in the range of 20 ° C. to 50 ° C. For example, preferably 25 ° C to 45 ° C, especially 30 ° C
A neighborhood of 27 ° C to 37 ° C is preferred. Subsequently, the pH conditions are from 6 to 12, preferably from 7 to 11, and especially from around 10 to 8 to
10.5 is preferred. Further, it is desirable to stir the treatment liquid in order to promote the reaction in the decomposition / solubilization treatment tank uniformly. The stirring conditions may be any conditions under which the decomposition / solubilization treatment in the present invention can be suitably performed, and can be set as appropriate at any time according to the shape of the reaction tank, the load of the jellyfish, and the like.

The method of crude extraction of a useful substance in the third step is carried out by any method as long as the target useful substance can be efficiently crudely extracted from the jellyfish solubilized solution decomposed and solubilized in the second step. For example, salting out, precipitation with an organic solvent, isoelectric precipitation, precipitation with a water-soluble polymer, centrifugation, ultrafiltration, membrane filtration, ion exchange chromatography, adsorption chromatography, distribution Methods such as chromatography, gel filtration chromatography, affinity chromatography, and isoelectric focusing can be appropriately selected depending on the extraction target, processing conditions, and the like.

The method for extracting useful substances derived from aquatic organisms according to the present invention is characterized by comprising the above three steps. FIG. 1 shows a flowchart of the steps of the method for extracting a useful substance derived from aquatic organisms of the present invention. 1 is a breaking / shredding step, 2 is a decomposing / solubilizing step, 3 is a rough extraction step of a useful substance, and 4 is an extraction of a useful substance. The present invention is carried out in the order of the above steps.

[0027]

The invention can be better understood by describing the following illustrative but non-limiting examples. (Example 1) A processing apparatus as shown in FIG. 2 was manufactured and tested. 4 is a stirring blade motor, 21 is a jellyfish crusher, 22 is a decomposition / solubilization tank, 23 is a coarse extraction device (ultrafiltration device), 25 is a heating device, 26 is a liquid sending pump,
7 is a valve. A treatment tank having an inner volume of 3m ³ of water was added 1 m ^3, after forming a fragment of Aurelia by crushing and shredding through the crushing and shredding device Aurelia 200 kg, 25
The stirring treatment was performed at a constant temperature of 120 ° C. and a constant temperature of 120 ° C.

After 15 hours, the treatment liquid was filtered, and the weight of the residue on the filter paper was measured. Table 1 shows the results. Here, protease activity was measured for the treatment solution. Sampling was performed with n = 3, and the test was repeated three times. According to this embodiment, an enzyme as a useful substance, that is, an alkaline protease which is a kind of an enzyme for decomposing proteins of moon jellyfish can be released and extracted. A treatment tank (Comparative Example 1) internal volume 3m ³ of water was added 1 m ^3, was added as a Aurelia 200kg was minced. This was filtered and the residue weight on the membrane was measured. Table 1 shows the results. In addition, protease activity was measured for the filtrate. Table 1 shows the results.

After completion of the weight measurement, the residue was returned to the filtered solution, salted out using ammonium sulfate to precipitate the protein, and separated from the supernatant by centrifugation and collected. This was dried at 37 ° C. to perform a dehydration treatment.

After the dehydration treatment, the substance containing the recovered protein was suspended again in distilled water, dialyzed, and the protease activity was measured. As a result, no protease activity was detected in the protein thus obtained.
Sampling was performed at n = 2, and the test was repeated three times. (Comparative Example 2) 200 kg of moon jellyfish was added to a treatment tank having an internal volume of 3 m ³ to which 1 m ³ of water had been added, and shredded. The mixture was sterilized in an autoclave, and shaken at 120 ° C under a constant temperature of 25 ° C. After 15 hours, the treatment liquid was filtered, and the weight of the residue on the membrane was measured. Table 1 shows the results. In addition, protease activity was measured for the treatment solution. Table 1 shows the results. The test was performed with n = 2, and was repeated three times.

[Table 1] In Example 1 described above, the enzyme, which is a useful substance, can be significantly extracted. In other words, although some enzymes could be taken out by simply breaking water organisms in the past, the taken-out enzymes were set to the optimal conditions under which the enzymes act as in the present example, and were directly applied to tissues of sea organisms, for example, cells. It is thought that by acting on the protein to break down the cells of aquatic organisms, a large amount of enzymes trapped in the cells can be released. (Embodiment 2) In the following description of the embodiment, repetition of the same steps, the same conditions, the same apparatus, and the like as in the first embodiment will be avoided.

In the present example, more optimum conditions were searched for by changing the pH conditions under which the enzyme acts. The protease activity of the treatment solution of Example 1 was measured under different pH conditions.
Otherwise, the recovery of useful substances from marine organisms was attempted in the same manner as in Example 1. FIG. 3 shows the measurement results.

In the figure, 30 is the case where a 100 mM phosphate buffer was used, 31 is the case where a 50 mM potassium borate buffer was used, and 32 is the case where a 100 mM glycine sodium chloride sodium hydroxide buffer was used.

The test was performed with n = 2, and was repeated three times. (Example 3) In Example 3, various temperatures were used as conditions for activating the enzyme, and a temperature condition with the highest efficiency for decomposing proteins was searched for. And it carried out similarly to Example 1 except having performed the 2nd process on desirable temperature conditions. The heater 25 was used for setting the temperature.

The temperature for the decomposition and solubilization treatment is 25 ° C., 35
The same shaking treatment as in Example 1 was performed, except that the temperature was set at five temperature settings of ° C, 45 ° C, 55 ° C, and 65 ° C. After the shaking treatment, the protease activity of each treatment solution was measured.
FIG. 4 shows the results.

The sampling was performed at n = 5, and the test was repeated three times. (Example 4) Example 4 is an example in which the type of useful substance extracted from the final treatment solution extracted in Example 1 was collagen. The other points are the same as the first embodiment.

[0036] Inner volume 5 with 100 mL of sterile distilled water added
A crushed jellyfish (20 g) was added to a 00 mL Erlenmeyer flask, and shaken at 35 ° C. and 120 rpm for 5 hours.

As a result, the jellyfish was decomposed and solubilized.
The filtrate obtained by fractionating 50 mL of the solubilized solution with an ultrafiltration membrane with a molecular weight cutoff of 3 × 10 ⁵ was further subjected to a molecular weight cutoff of 1 × 10 ^{4 with} an ultrafiltration membrane to perform crude extraction of collagen. . As a result of attempting to extract collagen from the sample after ultrafiltration, it was possible to extract collagen of up to about 3% by weight of jellyfish weight. The extraction scheme is shown in FIG. (Comparative Example 3) Collagen was extracted from 10 g of jellyfish by a conventional method. As a result, collagen of up to about 2.8% by weight of the jellyfish weight was extracted. The extraction scheme is shown in FIG. 50 shows a flowchart of a collagen extraction process in Comparative Example 5, and 51 shows a washing step, 5
2 is a defatting step, 53 is a pepsin digestion step, 54 is a salting-out step, 55 is a dialysis step using a buffer, and 56 is a dialysis step using acetic acid. 60 is a flow chart of the collagen extraction process in Example 5, 61 is a delipidation step, 62 is a pepsin digestion step, 63 is a salting-out step, 64 is a dialysis step using a buffer, and 65 is acetic acid. Each shows the dialysis process using.

From the results of Example 1 and Comparative Examples 1 and 2, and from the results of Example 4 and Comparative Example 3, if the method for extracting useful substances derived from aquatic organisms of the present invention is used, It can be easily understood that the efficient use of protease makes it possible to extract crude jellyfish useful substances more efficiently than conventionally proposed techniques. Further, from the results of Examples 2 and 3, the suitable processing condition range of the present invention can be easily understood.

[0039]

As described above, according to the present invention, the efficiency of extraction of a useful substance can be improved by reducing the unreacted portion, and a valuable added substance derived from aquatic organisms can be recovered.

[Brief description of the drawings]

FIG. 1 is a flowchart showing steps of a method for extracting a jellyfish-derived useful substance according to the present invention.

FIG. 2 is a schematic view of a test apparatus used in Example 1 of the present invention.

FIG. 3 is a graph showing preferable results in Example 2 of the present invention.

FIG. 4 is a graph showing preferable results in Example 3 of the present invention.

FIG. 5 is a flowchart of collagen extraction processing in Example 5 and Comparative Example 4 of the present invention.

[Explanation of symbols]

 4 Stirring blade motor 21 Jellyfish crusher 22 Decomposition / solubilization tank 23 Coarse extraction device (Ultrafiltration device) 25 Heating device 26 Liquid feed pump 27 Valve

 ──────────────────────────────────────────────────続 き Continued on the front page F term (reference) 4B029 AA02 DA01 DB01 4B050 CC01 DD11 LL05 4B064 AG01 CA21 CD21 CE02 DA16 4D056 AB12 AB20 AC22 BA08 CA06 CA22 DA01

Claims (3)

[Claims] 1. A first step of crushing and shredding aquatic organisms into fragments to release enzymes from the aquatic organisms, and the enzyme acting on proteins in the fragments to decompose the fragments into liquid substances A method for extracting useful substances from aquatic organisms, comprising: a second step of extracting useful substances from the liquid substance. 2. The method for extracting useful substances of aquatic organisms according to claim 1, wherein the enzyme is an alkaline protease. 3. The method according to claim 1, wherein the useful substance is collagen.