JP2021031444A - Extract - Google Patents

Abstract

To provide extract capable of containing balenine in abundance.SOLUTION: One extract of the present invention contains: proteolytic enzyme degradation products of fish meat, fish head, fish internal organs, fish skin, fish tail, or fish bone; and a balenine soluble in a physiologically acceptable liquid medium, or a balenine extractable from a physiologically acceptable liquid medium. Since the extract contains abundant amounts of balenine together with the proteolytic enzyme degradation products mentioned above, an antioxidant effect and/or an anti-fatigue effect and the like can be exerted by ingesting the extract.SELECTED DRAWING: Figure 1

Description

The present invention relates to extracts, especially those containing ophidine.

Health enriches the lives of citizens and contributes to the construction of a vibrant society. Providing foods that can help maintain or improve health is now an important issue not only in developed and developing countries. One of the representatives of such foods is marine products represented by fish. Humans have been eating fish for a long time, and their characteristics such as high-quality animal protein, low calories, and improvement of brain function have been attracting attention for a long time.

In recent years, among marine products that are said to enhance human physiological functions, fish focusing on antioxidant action and / or anti-fatigue action have also attracted attention (Non-Patent Document 1).

For example, there are mainly three types of substances classified as imidazole dipeptides, and in general, the types of imidazole dipeptides present in fish are biased. Specifically, among the imidazole dipeptides, anserine and carnosine are mainly contained in fish. For example, high-speed migratory fish such as bonito and tuna are known to contain a relatively large amount of anserine. In addition to seafood, carnosine and anserine are abundant in, for example, pork or chicken. Therefore, research on carnosine and anserine is being actively pursued. In particular, regarding carnosine, a composition for inhibiting carnosine dipeptidase is disclosed because carnosine is decomposed by carnosine dipeptidase present in serum or tissue and impairs the functioning of carnosine (Patent Document 1). ).

In addition, the present inventor found that lampris guttatus (also called "mandai") contains ophidine, which is a kind of imidazole dipeptide, which is more than the marine mammal minke whale, and discloses the fact. (Non-Patent Document 3, Non-Patent Document 4).

International Publication No. WO2017 / 104777

Nishitani, 3 others, "Review New Anti-Fatigue Ingredient: Imidazole Dipeptide", Journal of Japan Society for Complementary and Alternative Medicine, Vol. 6, No. 3, October 2009, p123-129 Takahashi, et al., "Effect of anserine-containing salmon extract on high-fat diet-fed rats", Journal of the Japanese Society of Fisheries Science, Vol. 76, No. 6, 2008, p1075-181 Omura, 8 others, "Valenin content in the muscle of Lampris guttatus", Proceedings of the 2018 Spring Meeting of the Japanese Society of Fisheries Science, March 26, 2018, p92 Omura, 9 others, "Discovery that lampris guttatus has a high content of ophidine that has a fatigue-suppressing function", Research Movement (National Research Institute of Fisheries Science), September 2018, p11

Among the imidazole dipeptides, ophidine is a substance that has not received much attention so far. This is because the animals containing ophidine in their muscles and the like are extremely limited, such as whales and scallops, which are marine mammals as described above. In addition, whale meat, known as ophidine-rich meat along with other imidazole dipeptides, is impossible or very difficult to ingest in situations where whaling is not easy. Therefore, even in a country that was once a whaling country, there are very few opportunities to take ophidine at present.

However, among the imidazole dipeptides, ophidine has antioxidant and / or anti-oxidant effects compared to other imidazole dipeptides (carnosine, anserine) due to the ecology of whales that can continue to swim long distances without eating food. It is a substance that is considered to have an advantage in terms of fatigue action. Also, therefore, it is extremely important for societies to maintain or improve their health to continue their efforts to find ways to continuously ingest imidazole dipeptides, especially ophidine, even in situations where whaling is not easy. ..

As described above, the present inventor has found that lampris guttatus contains ophidine in spite of fish, but how to extract the ophidine from the lampris guttatus makes the best use of the ophidine. Since it has not yet been examined whether it will be, it is necessary to examine and analyze it from various viewpoints. Therefore, it can be said that the research and development and materialization of the extraction of ophidine from fish have just begun.

The present invention greatly contributes to the realization of an extract that can be rich in ophidine.

Based on the situation that whale meat is difficult to ingest easily, the present inventor has conducted research and analysis including the ecology of marine organisms that may contain ophidine that replaces or exceeds whales. As a result of many investigations and analyzes, the present inventor has exerted an antioxidant effect and / or an anti-fatigue effect in the body by some influence in addition to the viewpoint of a migratory fish that can cope with continuous exercise. We have also come to pay attention to fish that are thought to be deaf, especially deep-sea fish.

Subsequent trial and error and analysis by the inventor revealed that the deep-sea fish Lampris guttatus (also called "mandai") contains abundant amounts of imidazole dipeptide, especially ophidine. I got it. Therefore, the present inventor has intensively studied and analyzed lampris guttatus, which is a kind of fish. As a result, we obtained a very interesting technical finding that the ophidine content does not change substantially even when heat is applied to almost all parts of lampris guttatus, for example.

In addition, among imidazole dipeptides, ophidine has antioxidant, anti-fatigue, active oxygen scavenging, autonomic regulation, alcohol metabolism promoting, blood glucose elevation, uric acid lowering, iron absorption, and eye strain. The present inventor has found that at least one selected from the group of reducing, promoting wound healing, metal chelating, and suppressing the decrease in muscle activity caused by pH decrease due to lactic acid can be exhibited.

Further analysis revealed that the imidazole dipeptides contained in the edible part (ordinary muscle) of lampris guttatus and the parts different from the edible part have the following characteristics (1) to (5). The inventor has learned.

(1) The parts (head, internal organs, and skin) of Lampris guttatus that are different from the so-called edible part (normal muscle) also contain an amount of ophidine comparable to that of the edible part (normal muscle).
(2) The amount of imidazole dipeptide contained in the part (visceral and skin) of Lampris guttatus different from the so-called edible part (ordinary muscle) is larger than the amount of imidazole dipeptide contained in the edible part (ordinary muscle). Remarkably many things.
(3) The amount of imidazole dipeptide contained in the so-called edible portion (ordinary muscle) of lampris guttatus and the imidazole dipeptide contained in a site (particularly internal organs and skin) different from the edible portion (ordinary muscle). The amount of each type is significantly different.
(4) The amount of imidazole dipeptide (including ophidine) in the bloody meat varies greatly (standard deviation) compared to the normal muscle of Lampris guttatus.
(5) The bone and tail 90 contain ophidine.

Furthermore, the present inventor has focused on the existence of an enzyme that appropriately decomposes a protein containing collagen, which is a main component of the fish meat, in order to efficiently extract the so-called ophidine buried in the fish meat. As a result of research and analysis for the extraction of ophidine by enzymatic decomposition of fish meat protein, the present inventor has found that ophidine can be efficiently extracted by adopting a certain enzyme.

The present invention has been created based on the above-mentioned viewpoints and technical knowledge.

One extract of the present invention is a proteolytic enzymatic degradation product of fish meat, fish head, fish internal organs, fish skin, fish tail, or fish bone, and ophidine, which is soluble in a physiologically acceptable liquid medium, or Contains ophidine, which can be extracted from a physiologically acceptable liquid medium.

Since this extract contains abundant amount of ophidine together with the above-mentioned proteolytic enzyme degradation product, by ingesting the extract, for example, antioxidant action, anti-fatigue action, active oxygen scavenging action, autonomy Neuromodulatory action, alcohol metabolism promoting action, blood glucose level rise suppressing action, uric acid level lowering action, iron absorption promotion, eye strain reduction, wound healing promotion action, metal chelating action, and decrease in muscle activity due to pH lowering due to lactic acid At least one selected from the group of inhibitory effects can be played.

By the way, in the present application, the "skin" or "skin portion" means a portion having a thickness of less than 10 mm from the outermost surface of a fish (for example, Lampris guttatus) toward the inside (inside the body). Further, in the present application, the “bloody meat” refers to a part of the muscle on the body side of the fish that exhibits a dark red color. Looking at another index, in the present application, the “bloody meat” refers to a portion containing an iron content of twice or more the iron content contained in the “ordinary muscle”.

According to one extract of the present invention, for example, antioxidant action, anti-fatigue action, active oxygen scavenging action, autonomic nerve regulation action, alcohol metabolism promoting action, blood glucose level rise suppressing action, uric acid level lowering action, iron absorption promoting action, At least one selected from the group of eye strain reducing action, wound healing promoting action, metal chelating action, and suppressing action of reducing muscle activity caused by pH lowering due to uric acid can be exhibited.

It is a flow which shows the manufacturing process of the extract of 1st Embodiment. It is a photograph which shows the cut part of the lampris guttatus of the first embodiment, and a part of the lampris guttatus showing the part. It is a flow which shows the manufacturing process of the extract of 2nd Embodiment.

Embodiments of the present invention include proteolytic enzymatic degradation products of fish meat, fish head, fish internal organs, fish skin, fish tail, or fish bone, and ophidine, or physiologically soluble in a physiologically acceptable liquid medium. An extract containing ophidine extractable from an acceptable liquid medium and a method for producing the same will be described in detail with reference to the accompanying drawings.

<First Embodiment>
FIG. 1 is a flow showing a manufacturing process of the extract of the present embodiment. More specifically, FIG. 1 shows proteolytic enzymatic degradation products of fish meat, fish head, fish internal organs, fish skin, fish tail, or fish bone, and ophidine soluble in a physiologically acceptable liquid medium. Alternatively, it is a flow showing a production process of an extract containing ophidine, which can be extracted from a physiologically acceptable liquid medium. In the extract of the present embodiment and the method for producing the same, each treatment step shown in FIG. 1 can be all steps or a part thereof in the method for producing the extract. Therefore, the method for producing the extract of the present embodiment does not necessarily include all of the treatment steps shown in FIG. Further, FIG. 2 is a photograph showing a cut portion of the lampris guttatus 100, which is an example of the fish of the present embodiment, and a part of the lampris guttatus 100 showing the portion.

(Freezing process and cutting process)
Specifically, as shown in FIG. 1, the caught lampris guttatus 100 is frozen (step S1). After that, as shown in FIG. 2, the head 10, the muscle 20 (more specifically, the normal muscle 20a and the bloody meat 20b), the internal organ 30, the bone (not shown), and the tail 90 of the lampris guttatus 100, respectively. A cutting (disassembling) step is performed in which the frozen lampris guttatus 100 is cut along _{C 1} , C ₂ , and C ₃ substantially linearly so as to be separated (step S2). In FIG. 2, a part of the part including the dorsal fin is not shown in the photograph, but the part not shown is not the raw material of the extract of the present embodiment, and thus the description of the part will be omitted.

Here, cutting as shown in C ₁ in FIG. 2, as sickle of red sunfish 100 of the present embodiment is not included in the muscle 20, and along a substantially tangential direction of the end portion of the muscle 20 side of the hook Will be implemented. Further, the cutting shown in C ₂ in FIG. 2, the eyes and mouth of the red sunfish 100 of the present embodiment is performed as included in the head 10. In addition, the cutting shown in C _3, the muscles 20 of red sunfish 100 of this embodiment is carried out as much as possible, not included in the tail 90. Further, the bone not shown in FIG. 2 is separated from other parts by a cutting process using a knife such as a kitchen knife or a cutting device such as a known electric grinder. Although a known band saw device is used in the cutting (disassembling) step of the present embodiment, the means for cutting the lampris guttatus in the step is not limited to the band saw device. Other known cutting methods capable of cutting frozen lampris guttatus along C ₁ , C ₂ , and C _{3 may also be employed.}

After that, in order to perform fragmentation by a known chopper device (for example, manufactured by Natsune Co., Ltd., model: MC-22), the red was frozen to the extent that it could be introduced into the chopper device. Each part of the sunfish 100 (head 10, normal muscle 20a, bloody meat 20b, internal organs 30, skin 40, bone, tail 90) is thawed completely or incompletely, respectively. Then, by performing the shredding step using the chopper device, for example, the minced minced parts can be obtained from a plate having a through hole of 10 mmφ. Although a known chopper device is used in the shredding step of the present embodiment, the means for obtaining the shredded parts in the step is not limited to the chopper device. For example, known silent cutters can also be employed in the shredding process.

[Analysis of ophidine content extracted from each part of fish without using enzymes]
Here, the present inventor analyzed the amount of imidazole dipeptide (particularly ophidine) contained in each of the fragmented sites of the present embodiment. Specifically, after preparing a perchloric acid extract from each of the fragmented parts as described above, each part of the fragmented lampris guttatus 100 is prepared using a known amino acid automatic analyzer. Analysis of free amino acids and bound amino acids was performed on (head 10, normal muscle 20a, bloody meat 20b, visceral part 30, skin part 40, bone, tail part 90).

In addition, "Bal." In Table 1 means "content of ophidine". The skin 40 as the analysis target in Table 1 is the skin of the muscle 20. Each numerical value is expressed as the amount of ophidine (the amount of acquired ophidine) with respect to the fish meat (ordinary muscle 20a) of Lampris guttatus 100 as mass% (wt%).

Moreover, in this analysis, the presence of ophidine was clarified based on the result that a high concentration of 3-methylhistidine was detected by hydrolyzing with hydrochloric acid and analyzing the bound amino acid. In addition, the amount of ophidine in each fragmented site was quantified by quantifying the amount of ophidine based on 3-methylhistidine in a hydrolyzed solution using a standard ophidine product. The method for quantifying the amount of ophidine is the same as the method for quantifying the amount of ophidine in the extraction step by enzymatic decomposition, which will be described later.

As shown in Table 1, the above-mentioned analysis revealed the following characteristic (1) regarding the amount of ophidine in each fragmented site.

(1) The parts (head, internal organs, and skin) of Lampris guttatus that are different from the so-called edible part (normal muscle) also contain an amount of ophidine comparable to that of the edible part (normal muscle).

In addition, although not shown in Table 1, as a result of analysis by the present inventor, the following findings (2) to (5) were also obtained.
(2) The amount of imidazole dipeptide (combined with ophidine, anserine, and carnosine) contained in the part (visceral and skin) different from the so-called edible part (ordinary muscle) of lampris guttatus is the edible part ( Significantly higher than the amount of imidazole dipeptide contained in normal muscle).
(3) The amount of imidazole dipeptide contained in the so-called edible portion (ordinary muscle) of lampris guttatus and the imidazole dipeptide contained in a site (particularly internal organs and skin) different from the edible portion (ordinary muscle). The amount of each type is significantly different.
(4) The amount of imidazole dipeptide (including ophidine) in the bloody meat varies greatly (standard deviation) compared to the normal muscle of Lampris guttatus.
(5) The bone and tail 90 contain ophidine.

More specifically, the above-mentioned feature (1) will be described more specifically. At least each fragmented part (head 10, normal muscle 20a, bloody meat 20b, visceral part 30) derived from Lampris guttatus 100 is contained. It can be said that the amount of ophidine to be produced is equal to or considerably larger than the amount of ophidine contained in the red meat of the back meat of minke whales. Interestingly, among the normal muscles 20a in Table 1, the ophidine content and the imidazole dipeptide content of the normal muscle 20a from the center to the cranial side of the normal muscle 20a are the normal muscles from the center to the caudal side of the normal muscle 20a. It was confirmed that it was about 6 wt% or more higher than the content of ophidine and imidazole dipeptide of 20a.

In the present embodiment, the position of the "center of the normal muscle 20a" is the length from the tip of the head 10 (the tip of the mouth) to the caudal end of the normal muscle 20a (thus not including the tail 90). It is defined as a position 0.5 from the cranial tip when the (“L” in FIG. 2) is 1. Therefore, in order to obtain the normal muscle 20a having a high content of valenin with higher accuracy, it is preferable to obtain the imidazole dipeptide and / or valenin from the normal muscle 20a including the normal muscle 20a from the cranial tip to 0.4 or less. This is one aspect.

(Extraction process by enzymatic decomposition)
In the present embodiment, after the cutting step is performed, typically, the fragmented normal muscle 20a of Lampris guttatus 100 is used as an extraction medium (liquid medium) at about 50 ° C. containing a certain proteolytic enzyme. Contact with water (ion-exchanged water). More specifically, an extraction step (step S3) is performed in which the fragmented ordinary muscle 20a is immersed in the extraction medium at about 50 ° C. in a known container. As a result, an extract of the present embodiment containing a proteolytic enzyme degradation product and ophidine, which will be described later, is obtained. In the extraction step of the present embodiment, it is also possible to intentionally stir the extraction medium.

The contact time in the extraction step of this embodiment is about 2 hours. In addition, the types of enzymes adopted in this embodiment are the following three types (e-1) to (e-3).
(E-1) An enzyme having both an endopeptidase action and an exopeptidase action, or an enzyme in which an enzyme having an endopeptidase action and an enzyme having an exopeptidase action are mixed (e-2) An enzyme having an endopeptidase action (Cysteine protease)
(E-3) Enzyme having endopeptidase action (serine protease)

Here, the example of the enzyme shown in (e-1) above may be either one type of enzyme or a mixture of two or more types of enzymes. Further, among the enzymes shown in (e-1) above, an example of an enzyme having both an endopeptidase action and an exopeptidase action is Streptomyces griseus-produced peptidase, which has an endopeptidase action. An example of an enzyme that has is a serine protease. Examples of the enzyme having an exopeptidase action include an enzyme having an exopeptidase action contained in the product name "Denateam AP" manufactured by Nagase ChemteX Corporation, or leucine aminopeptidase (LAP) or dipeptidyl-aminopeptidase (. DAP) can be adopted.

An example of the above-mentioned enzyme (cysteine protease) having an endopeptidase action (e-2) is papain. Further, an example of the enzyme (serine protease) having an endopeptidase action (e-3) described above is peptidase produced from Bacillus subtilis.

(Separation process)
Then, in the present embodiment, about 2 hours after the fragmented normal muscle 20a is immersed in the extraction medium, an extract from the fragmented normal muscle 20a (in the present embodiment). , Aqueous solution) is separated from the normal muscle 20a (which can be treated as a residue) using a centrifuge (step S4). Specifically, the extract of the present embodiment can be produced by performing a centrifugal filtration treatment using a 300-mesh filter cloth. The extract of this embodiment is liquid at this stage. Further, in the separation step of the present embodiment, the centrifugal filtration treatment using a filter cloth of 300 mesh is adopted, but the separation step of the present embodiment is not limited to such an embodiment. It is another aspect in which a known separation step (for example, separation treatment by centrifugal sedimentation) can also be adopted. Further, in the present embodiment, the normal muscle 20a is immersed in the extraction medium for about 2 hours, but the immersion time can be appropriately changed depending on the purpose of extracting ophidine.

In the present embodiment, the normal muscle 20a has been taken up and described as a representative, but the present embodiment is not limited to the normal muscle 20a. For example, the present embodiment also relates to an extract containing at least one proteolytic enzyme degradation product selected from a group of fish meat, fish head, fish internal organs, fish skin, fish tail, and fish bone, and ophidine. Can play at least part of the effect of.

<Modified example (1) of the first embodiment>
In this modification, as shown in FIG. 1, by further concentrating the extract produced by the process of producing the extract of the first embodiment, the extract as a concentrated extract containing valenin (hereinafter referred to as “extract”). , Simply referred to as "concentrated extract") can be performed in a concentration step (step S5).

In the concentration step (step S5) of this modification, for example, the first implementation is carried out in a space in a reduced pressure (for example, 0.03 MPa) state at a temperature of about 60 ° C. for about 1 hour to about 8 hours. A concentrated extract can be produced by stirring or allowing the morphological extract to stir.

By the way, in the concentration step of this modification, the above-mentioned processing conditions are adopted, but the concentration step of this modification is not limited to such an embodiment. It is another aspect in which another known concentration step different from the purification step described later can also be adopted. Further, it is another aspect that can be adopted that a sterilization treatment (for example, a heat treatment) is performed before the concentration step of the present modification is performed. Further, in the concentration step (step S5), if the temperature at which the extract of the first embodiment is stirred or allowed to stand is 20 ° C. or higher, 30 ° C. or higher, 40 ° C. or higher, or 50 ° C. or higher, this modification At least part of the effect of can be achieved.

<Modified example (2) of the first embodiment>
In this modification, as shown in FIG. 1, the extract of the first embodiment in which the above-mentioned concentration step (step S5) is not performed is dried, or the modification of the first embodiment (1). ), The drying step (step S6) of obtaining the dried extract can be carried out by drying the concentrated extract produced by the manufacturing step of the extract as the concentrated extract.

In this modification, a vacuum drying (including vacuum drying) step or a freeze-drying step is performed as a typical example of the drying step. Here, the "vacuum drying (or vacuum drying) step" in this modification means a step of drying in a vacuum state (or a reduced pressure state) while heating. Further, the "freeze-drying step" in this modification includes a "freeze-vacuum drying step" or a "freeze-vacuum drying step".

As long as the amount of imidazole dipeptide (particularly valenin) contained in the dried extract of this modified example is not significantly impaired (for example, about 20 wt% or more), hot air drying is performed instead of the above-mentioned two types of drying steps. , Microwave drying, superheated steam drying, frying drying, vacuum frying drying, drum drying, or sun drying can also be adopted.

For example, when the vacuum drying step is adopted, known methods and devices can be used. For example, a continuous belt dryer or a vacuum drum dryer may be adopted. In the present embodiment, a modified example (1) of the first embodiment is performed for about 12 hours to about 72 hours under a pressure of −0.093 MPa with a gauge pressure of −0.093 MPa with the atmospheric pressure as 0 and in an environment of about 60 ° C. ) Can be stirred or allowed to stand to form a dry extract of this variant. In this vacuum drying step, the temperature at which the concentrated extract of the modified example (1) of the first embodiment is stirred or allowed to stand is 20 ° C. or higher, 30 ° C. or higher, 40 ° C. or higher, or 50 ° C. or higher. For example, at least a part of the effect of this modification can be achieved.

Next, when the freeze-drying step is adopted, known methods and devices can be used. For example, a freeze-drying device (manufactured by Kyowa Vacuum Technology Co., Ltd., model: RL4522BS) can be adopted. In the present embodiment, the concentrated extract of the modified example (1) of the first embodiment is stirred or allowed to stand for about 168 hours under a pressure of about 266 Pa and an environment of about -20 ° C. Allows the formation of a dry extract of this variant.

The water content of the dried extract of this modified example represented by vacuum drying or freeze drying can be appropriately adjusted by a known means. However, from the viewpoint of ease of transportation and / or ease of handling, it is a preferable aspect that the water content is more than 0 wt% and 10 wt% or less of the dried extract of this modified example. From the above viewpoint, the water content is more preferably 5 wt% or less of the dry extract of the present modification, and further preferably 3 wt% or less of the dry extract of the present modification.

To obtain a crude powder by further pulverizing the dried extract obtained by drying the extract of the first embodiment in which the above-mentioned concentration step (step S5) is not performed, or the dried extract of the present modification. It is one aspect that can be adopted that the powdering step (step S7) is performed. In this embodiment, for example, a known hammer mill (for example, a hammer mill manufactured by Hosokawa Micron Co., Ltd.) can be adopted. More specifically, the dried extract of the present modification can be pulverized under the treatment conditions set so that the particle size is 2 mm or less.

As long as the amount of imidazole dipeptide (particularly ophidine) contained in the dried extract of this modified example is not significantly impaired (for example, about 20 wt% or more), instead of the above-mentioned hammer mill, a pin mill, a ball mill, or an impact Mills, edge runner mills, roller mills, vibration mills, conical grinders, jet mills, or other known mechanical impact / grinders can also be employed.

Through each of the above steps, a dry extract (hereinafter, also simply referred to as “crude powder”) as a crude powder of the present embodiment, which is derived from Lampris guttatus 100, can be produced.

The water content of the dry extract as the crude powder of this modification can also be adjusted as appropriate by known means in the same manner as described above. However, from the viewpoint of ease of transportation and / or ease of handling, it is a preferable aspect that the water content is more than 0 wt% and 10 wt% or less of the crude powder of this modified example. From the above viewpoint, the water content is more preferably 5 wt% or less of the crude powder of the present modified example, and further preferably 3 wt% or less of the crude powder of the present modified example.

In addition, the crude powder of this modification is a crude powder produced without going through a purification step. More specifically, the crude powder is prepared, for example, as in the method disclosed in Non-Patent Document 2, after the crude powder is dissolved in water or the like, and then an ion exchange resin (for example, cation exchange) is used. It is a crude powder obtained without performing a step of extracting and purifying an imidazole dipeptide containing ophidine using a resin). Therefore, even if it is an unpurified crude powder, for example, antioxidant action, anti-fatigue action, active oxygen scavenging action, autonomic nerve regulation action, alcohol metabolism promoting action, blood glucose level increase suppressing action, uric acid level lowering action, iron An imidazole dipeptide capable of exerting at least one selected from the group of absorption promotion, eye fatigue reduction, wound healing promotion action, metal chelating action, and suppression action of decrease in muscle activity due to pH decrease due to lactic acid, particularly The abundant amount of valenin as described above improves the yield (including yield) of imidazole dipeptide and / or valenin, simplifies the manufacturing process (including shortening of manufacturing time), and manufactures. It deserves special mention because it creates many advantages such as cost reduction.

[Analysis of ophidine content extracted after extraction process by enzymatic decomposition]
After obtaining the dry extract as the crude powder described above, the present inventor, in particular, paying attention to the amount of valenin obtained from the extract, perchloric acid extraction as in the first embodiment. After preparing the extract, analysis on free amino acids and bound amino acids was performed using a known automatic amino acid analyzer. Tables 2 and 3 show the analysis results. In Table 3, the unit of each numerical value in Table 2 is replaced with mass% (wt%) (however, in order to make the table easier to see, "about" is omitted in each numerical value in Table 3. .).

As shown in Tables 2 and 3, the amount of ophidine (the amount of ophidine obtained) is 1 wt% with respect to the fish meat (normal muscle 20a) of Lampris guttatus 100 regardless of which proteolytic enzyme is adopted. It was confirmed that the above (more narrowly, 1.4 wt% or more) can be obtained. Further, if the proteolytic enzyme (e-1) or (e-2) is adopted, the amount of ophidine is 2 wt% or more (2.5 wt in a narrower sense) with respect to the fish meat (normal muscle 20a) of Lampris guttatus 100. It was confirmed that it is possible to obtain% or more, more narrowly 2.9 wt% or more). In particular, when the proteolytic enzyme is (e-1), it was confirmed that the amount of ophidine can be 3 wt% or more with respect to the fish meat (normal muscle 20a) of Lampris guttatus 100.

Therefore, the extract containing the proteolytic enzyme-degraded product and valenin obtained by using the proteolytic enzyme has ophidine (the amount of ophidine obtained) when the mass ratio of normal muscle fish meat is 100. It was revealed that it contained 1 or more (1.4 or more in a narrower sense, 2 or more in a narrower sense, 2.9 or more or 3 or more in a very narrow sense).

As a result of further analysis of the enzyme for extracting ophidine by the present inventor, very interesting findings were obtained. Specifically, when at least the fish meat of Lampris guttatus is immersed in water, the pH value of the aqueous solution containing the extract of each part of Lampris guttatus is 7 or less (less than 7 in a narrower sense, and more narrowly, It was confirmed that it would be 6.5 or less). Thus, the liquid extract obtained by immersing the lampris guttatus fish meat, fish head, fish guts, fish skin, fish tail, or fish bone in water will be neutral or acidic.

Therefore, for example, when Akamanbou is adopted among fish for acquisition by extraction of ballenin using an enzyme, the extraction solvent or the extracted liquid extract is acidic or neutral water or an aqueous solution (hereinafter referred to as “Aqueous solution”). In this modified example, the term "aqueous solution") is a preferred embodiment from the viewpoint of obtaining more valenin. In other words, the above-mentioned first containing a proteolytic enzymatic degradation product by an enzyme capable of exerting higher activity in an aqueous solution having a pH value of more than 7 than in an aqueous solution having a pH value of 7 or less or less than 7. The embodiment of, or the extract of each of the above or below variants, will contain a higher amount of ophidine.

Focusing on the amount of ophidine obtained from fish meat (typically, normal muscle 20a) in Tables 1 and 2, the extraction when the proteolytic enzyme (e-1) or (e-2) was adopted. It can be seen that the substance can contain a larger amount of ophidine than the extract when extracted without the proteolytic enzyme (e-1) or (e-2). This indicates that the proteolytic enzyme can efficiently extract the ophidine that is buried in the fish meat.

Since the proteolytic enzyme (e-3) in the present embodiment was a proteolytic enzyme whose activity was enhanced in an alkaline aqueous solution, it is considered that the amount of ophidine obtained was smaller than that of other enzymes. Therefore, more ophidine can be obtained by adopting (e-3) a proteolytic enzyme whose activity is enhanced in an acidic or neutral aqueous solution among enzymes having an endopeptidase action (serine protease).

By the way, in the above-mentioned first embodiment, or in each of the above-mentioned or later modified examples, the amount of ophidine obtained from fish meat (typically, ordinary muscle 20a) is described, but the above-mentioned first embodiment is described. The embodiment of, or each of the above-mentioned or later-described modifications, can also be applied to the amount of ophidine obtained from a site other than fish meat or normal muscle 20a.

Further, in the above-mentioned first embodiment, or in each of the above-mentioned or later-described modifications, the extraction medium was water, but in the above-mentioned first embodiment, or in each of the above-mentioned or later-described modifications, the extraction medium was water. , The extraction medium is not limited to water. For example, adopting a mixed solution (aqueous solution) of water and alcohol (for example, ethanol) as an extraction medium (liquid medium) is also an embodiment of the above-mentioned first embodiment or each of the above-mentioned or later-described modifications. Is. The above-mentioned "alcohol" is not limited to ethanol. For example, the "alcohol" is brewed liquor containing beer, wine, and sake, distilled liquor containing whiskey, shochu, and vodka, and cassis and cassis unless the alcohol concentration is such that the effect of the present embodiment does not occur. May include mixed liquors, including Campari.

In addition, in the above-mentioned first embodiment, or in each of the above-mentioned or later modified examples, the extract obtained from fish meat (typically, ordinary muscle 20a) is described, but the above-mentioned first embodiment is described. The embodiment of the above, or each of the above-mentioned or later-described modifications, is an extract obtained from a part other than fish meat or normal muscle 20a (head 10, bloody meat 20b, internal organs 30, skin 40, bone, tail 90). Can also be applied to.

As described above, according to the first embodiment and each modification, by adopting one of the following production methods, an extract containing a proteolytic enzyme degradation product and a large amount of ophidine is obtained. Obtainable.
(Example of method for producing the extract (1))
Extraction medium (typically water) that is a physiologically acceptable liquid medium for fish meat, fish heads, fish guts, fish skins, fish tails, or fish bones in fish (typically Akamanbo). And / or aqueous solution) with proteolytic enzymatic degradation products of fish meat, fish head, fish guts, fish skin, fish tail, or fish bone, and valenin soluble in the extraction medium, or the extraction medium. A method for producing an extract, which comprises an extraction step containing, and, which can be extracted from.
(Example of method for producing the extract (2))
A method for producing an extract, further comprising a concentration step of concentrating the above-mentioned extract after the above-mentioned extraction step to obtain a concentrated extract containing the above-mentioned proteolytic enzyme degradation product and the above-mentioned ophidine.
(Example of method for producing the extract (3))
After the above-mentioned extraction step or the above-mentioned concentration step, a drying step of drying the extract so that the water content of the above-mentioned extract is more than 0 wt% and 10 wt% or less to obtain a dry extract is further added. Method of producing the extract, including.
(Example of method for producing the extract (4))
A method for producing an extract, further comprising a pulverization step of pulverizing the above-mentioned dry extract to obtain a powder of the dry extract having a particle size of less than 1 mm.

By the way, in the above-mentioned first embodiment and each modification, the enzymes shown in the above-mentioned (e-1) to (e-3) are adopted as the proteolytic enzyme, but the enzyme of the present embodiment The types are not limited to them. For example, other examples of enzymes with exopeptidase activity that can be employed in this embodiment are aminopeptidase, dipeptidase-peptidase, trypeptidyl-peptidase, peptidyl-zipeptidase, serine-type carboxypeptidase, metal carboxypeptidase, and / or. It is a cysteine type carboxypeptidase. In addition, other examples of enzymes having endopeptidase action that can be adopted in the present embodiment are serine endopeptidase, aspartic acid endopeptidase, metalloendopeptidase, threonine endopeptidase. In addition, lipid-related enzymes (lipase, phosphorlipase), plant tissue-disintegrating enzymes (cellulase, pectinase, hemicellulase), nucleic acid-related enzymes (endonuclease, adenylate deaminase), amino acid-related enzymes (asparaginase, glutaminase) can also be adopted. This is an example of an enzyme. As described above, even when an enzyme different from the proteolytic enzymes (e-1) to (e-3) is adopted, at least a part of the effect of this modification can be exhibited.

Further, as described above, among the fish meat, the amount of imidazole dipeptide (including ophidine) of the blood mixture 20b has a very large variation (standard deviation) as compared with the normal muscle 20a, so that it is formed from the blood mixture in the fish meat. The fact that the ratio of the obtained extract is less than 5 wt% has the effect of reducing the variation in the amount of ophidine contained in the extract obtained in the above-mentioned first embodiment or in each of the above-mentioned or later-described modifications. Can be played. In other words, the proportion of the extract formed from the bloody meat in the fish meat is less than 5 wt%, which can realize the production of the extract containing a more stable amount of ophidine.

<Modified example (3) of the first embodiment>
In this modification, as shown in FIG. 1, the dry extract as a crude powder produced by the production process of the modification (2) of the first embodiment is further classified to be less than 1 mm (more). A classification step (step S8) for obtaining a dry extract (hereinafter, also simply referred to as “fine powder”) as a fine powder composed of a powder having a particle size of (preferably less than 0.8 mm) can be performed.

By adopting the classification step (step S8) of this modification, for example, only fine powder passing through a sieve having a mesh opening of 0.7 mm can be recovered. As a result, a dry extract of the present modification (a dry extract as a fine powder composed of a powder having a particle size of less than 1 mm) can be produced. When the hammer mill or the like adopted in the powdering step in the modified example (2) of the first embodiment also has a classification function, the powdering step and the classification step can be carried out at the same time.

The water content of the dried extract as the fine powder of this modification can also be adjusted as appropriate by a known means in the same manner as described above. However, from the viewpoint of ease of transportation and / or ease of handling, it is a preferable aspect that the water content is more than 0 wt% and 10 wt% or less of the fine powder of this modified example. From the above viewpoint, the water content is more preferably 5 wt% or less of the fine powder of the present modified example, and further preferably 3 wt% or less of the fine powder of the present modified example.

As described above, the dry extract as the fine powder in this modification is a fine powder produced without undergoing a purification step. More specifically, the fine powder is prepared, for example, as in the method disclosed in Non-Patent Document 2, after the fine powder is dissolved in water or the like, an ion exchange resin (for example, a cation exchange resin) is used. It is a fine powder obtained without performing a step of extracting and purifying an imidazole dipeptide containing ophidine using the above. Therefore, even if it is an unpurified fine powder, for example, it has an antioxidant effect, an anti-fatigue effect, an active oxygen scavenging effect, an autonomic nerve regulation effect, an alcohol metabolism promoting effect, a blood glucose level increase suppressing effect, a uric acid level lowering effect, and iron absorption. An imidazole dipeptide capable of exerting at least one selected from the group of promotion, reduction of eye strain, promotion of wound healing, metal chelating action, and suppression of decrease in muscle activity due to pH decrease due to lactic acid, particularly abundant. It is worth noting that it contains a large amount of valenin.

<Second embodiment>
In the extract of the present embodiment and the method for producing the same, each treatment step shown in FIG. 3 can be all steps or a part thereof in the method for producing the extract. Therefore, the method for producing the extract of the present embodiment does not necessarily include all of the treatment steps shown in FIG. Moreover, the description overlapping with the first embodiment and each modification thereof may be omitted.

Specifically, as shown in FIG. 3, each step (step) from the freezing step to the drying step (steps S1 to S6) and from the freezing step to the pulverization step shown in the first embodiment and each modification thereof. This is carried out after S1 to S7) or each step (steps S1 to S8) from the freezing step to the classification step (not shown), or after the separation step (steps S1 to S4) is performed from the freezing step. The purification step of the form (step S9) is performed.

In the purification step (step S9) of the present embodiment, as in the method disclosed in Non-Patent Document 2, a dry extract or a dry extract as a crude powder is dissolved in water or the like, and then ion exchange is performed. A step of extracting and purifying an imidazole dipeptide containing valenin is performed using a resin (for example, a cation exchange resin).

Since the upper limits of the imidazole dipeptide content and the ophidine content with respect to the total mass of the extract obtained by performing the purification step (step S9) of the present embodiment are not particularly limited, the upper limits of the respective contents are not particularly limited. So to speak, it can be said that it is less than 100 wt%. If the upper limit is intentionally mentioned, in a narrower sense, the content of the imidazole dipeptide and the content of the ophidine are 99 wt% or less, and in a narrower sense, 95 wt% or less. Further, the numerical range of this upper limit value may be applied to other embodiments.

<Other Embodiments>
By the way, in the first embodiment and each modification thereof, and in the second embodiment, each part of the red manbow 100 (head 10, normal muscle 20a, bloody meat 20b, visceral part 30, skin part 40, bone). , Tail 90), individually extraction step (step S3) and separation step (step S4), and if necessary concentration step (step S5), drying step (step S6), pulverization step (step S6). Step S7), a classification step (step S8), or a purification step (step S9) is performed, but the present embodiment is not limited to such an embodiment.

For example, as an analysis by the present inventor, it was confirmed that the visceral part 30 and the skin part 40 have a large amount of imidazole dipeptide because they contain a large amount of carnosine. Therefore, it is also possible to adopt that each step after the cutting (disassembling) step (step S2) is carried out so that the internal organ portion 30 and the skin portion 40 can be mixed. In addition, all of the sites (head 10, normal muscle 20a, bloody meat 20b, internal organs 30, skin 40, bones, tail 90) having a remarkable feature of being rich in imidazole dipeptide containing ophidine. It is also an aspect that each step after the cutting (disassembling) step (step S2) is carried out so as to be in a state where the mixture can be mixed.

Further, in the first embodiment and its respective modifications, and in the second embodiment, the lampris guttatus caught by the longline fishing boat is promptly (for example, within 60 minutes) low-temperature freezing treatment (for example, within 60 minutes) on the fishing boat. , -65 ° C or higher and -15 ° C or lower), but the method of catching lampris guttatus and the method of preserving lampris guttatus on a fishing boat are not limited to the above examples. However, from the viewpoint of reducing the possibility of reducing the amount of imidazole dipeptide (typically, ophidine) by autolysis, the caught lampris guttatus is promptly (for example, within 60 minutes) on the fishing boat. Low temperature freezing treatment (for example, −65 ° C. or higher and −15 ° C. or lower) is a preferred embodiment.

Further, in the first embodiment, each part of the fragmented lampris guttatus 100 is immersed in an extraction medium (for example, water) containing a proteolytic enzyme at about 50 ° C. in the container. An extraction step (step S3) is performed, but the first embodiment is not limited to such an extraction step. For example, adopting the extraction step (step S3) as shown in the following (a) to (c) is also a suitable modification of the first embodiment.
(A) Akamanbou fragmented in an extraction medium (for example, water) having a temperature of 30 ° C. or higher, 40 ° C. or higher, 50 ° C. or higher, or 60 ° C. or higher and lower than the temperature at which the proteolytic enzyme is inactivated. Extraction step in which each part of 100 is immersed (b) The pressure in the container in the extraction step is different from the normally adopted pressure of about 1 atm (about 0.101 MPa) (for example, about 0.001 MPa or more and about 100 MPa or less). (However, excluding about 0.101 MPa)) Extraction step (c) The extraction medium is a physiologically acceptable liquid medium (eg, an organic liquid medium) and / or other physiologically acceptable extraction. Extraction step of salt water as an example of a medium (liquid medium) (preferably salt water having a low concentration (for example, salt water having a salt water concentration of less than 10 wt%). Here, the above-mentioned "physiologically acceptable liquid medium" is used. It goes without saying that it contains water.

When the extraction step (a) is adopted among the extraction steps (step S3) described as a modification of the first embodiment described above, extraction at a lower temperature is realized, which saves energy. Can contribute. As a result of investigation by the present inventor, even if the temperature of the extraction medium is 20 ° C. under about 1 atm (about 0.101 MPa) which is usually adopted, the temperature (about 50 ° C.) as an example of the first embodiment. ), It has been found that about 0.33 can be extracted when the amount that can be extracted is 1.

It should be noted that imidazole dipeptide (typically, valenin) can be extracted even with an extraction medium having an extraction temperature of 20 ° C., which is considerably low.

Further, in each extraction step (step S3) described as a modification of the first embodiment described above, when the extraction step (b) is adopted, for example, the pressure is higher than the pressure normally adopted. When extracted, the extraction step can be implemented using a cooler extraction medium.

Further, when the extraction step (c) is adopted among the extraction steps (step S3) described as a modification of the first embodiment described above, it can contribute to improving the flavor of the extract. ..

Further, in the first embodiment and its respective modifications (1) to (3), the second embodiment, and other embodiments, the dorsal fin of Akamanbou 100 can be used as a physiologically acceptable liquid medium. Extracts extracted using a soluble or physiologically acceptable liquid medium as an extraction medium (typically, but not limited to extracts, including concentrated extracts and dry extracts; , The same.), But the extract is not limited to such embodiments. For example, in order to further simplify the manufacturing process of the extract, without cutting shown in C ₃ in FIG. 2, it is an aspect that may be employed for each subsequent step. Therefore, it can also be adopted that the dorsal fin is included as one of the constituents of the extract of the first embodiment and its respective modifications (1) to (3), and the second embodiment.

Further, tablets and capsules containing a proteolytic enzyme degradation product and valenin as at least a part of the constituents produced in the first embodiment and its respective modifications, the second embodiment, and other embodiments. , Powders (including granules), pills, syrups, liquids, drinks, troches, drops, starch syrup, etc. (hereinafter collectively referred to as "formulations") are suitable to be adopted. This is one aspect. Since the preparation contains imidazole dipeptide, particularly abundant amount of ophidine, by ingesting the preparation, for example, antioxidant action, anti-fatigue action, active oxygen scavenging action, autonomic nerve regulation action, alcohol From the group of metabolism promoting action, blood glucose level rise suppressing action, uric acid level lowering action, iron absorption promotion, eye strain reduction, wound healing promotion action, metal chelating action, and suppression action of muscle activity decrease due to pH decrease due to lactic acid. At least one selected can be played. The preparation can be produced by a known production method. In addition, the preparation is appropriately mixed with an excipient, a binder, a disintegrant, a wetting agent, honey, rice flour, etc. in order to improve moldability or ease of oral administration. Can be formed.

Since one extract and preparation of the present invention is derived from fish that can be rich in ophidine, it is extremely useful not only in the food industry and the fishery industry but also in each industry related to pharmaceuticals, health, medical care, and beauty. ..

10 Head 20 Muscle 20a Normal muscle 20b Bloody meat 30 Internal organs 40 Skin 90 Tail 100 Lampris guttatus

Claims (8)

With proteolytic enzyme degradation products of fish meat, fish head, fish internal organs, fish skin, fish tail, or fish bone,
Containing ophidine soluble in a physiologically acceptable liquid medium or ophidine extractable from a physiologically acceptable liquid medium,
Extract. In terms of mass ratio, when the fish meat of normal muscle is 100, the ophidine is 1 or more.
The extract according to claim 1. The proteolytic enzymatic degradation product is a proteolytic enzymatic degradation product by an enzyme capable of exhibiting higher activity in an aqueous solution having a pH value of more than 7 than in an aqueous solution having a pH value of 7 or less or less than 7.
The extract according to claim 1 or 2. In terms of mass ratio, when the fish meat of normal muscle is 100, the ophidine is 2 or more.
The extract according to claim 1 or 3. The proteolytic enzyme degradation product is a proteolytic enzyme degradation product produced by an enzyme having both endopeptidase action and exopeptidase action, or a mixture of an enzyme having endopeptidase action and an enzyme having exopeptidase action. is there,
The extract according to any one of claims 1 to 4. The proteolytic enzyme degradation product is a proteolytic enzyme degradation product of the fish meat, and the proportion of the extract formed from the bloody meat in the fish meat is less than 5 wt%.
The extract according to any one of claims 1 to 5. A powder having a water content of more than 0 wt% and 10 wt% or less.
The extract according to any one of claims 1 to 6. From fish meat, fish heads, fish guts, fish skins, fish tails, or fish bone proteolytic enzyme degradation products and ophidine soluble in a physiologically acceptable liquid medium, or from a physiologically acceptable liquid medium Contains extractable ophidine, and,
Tablets, capsules, powders, pills, syrups, liquids, drinks, lozenges, drops, or starch syrup,
Formulation.