JP2696485B2 - Extraction method of fish and shellfish algae extract containing large amount of mineral components - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to pharmaceuticals, functional foods,
It is useful as a drug, various growth agents, feed additives for livestock, seasonings, etc., and mineral components such as phosphorus, calcium, zinc, iron and potassium contained in raw fish and shellfish algae, and large mineral components containing a large amount of taurine The present invention relates to a method for extracting a fish and shellfish algae extract.

[0002]

2. Description of the Related Art Various methods have been proposed for extracting a fish and shell algae extract by causing a protease to act on fish and shell algae, separating the extract into waste and waste. The resulting fish and shellfish algae extract contains effective peptides and free amino acids, and is used as various functional food additives, drugs, growth agents, livestock feed additives, seasonings, and the like. More specifically, for example, Japanese Patent Publication No.
In H9-10, a method of adding an alkali-resistant proteolytic enzyme to a raw fish to degrade the protein, adjusting the pH to 5-6, adding / decomposing the acid-resistant proteolytic enzyme, and concentrating the separated solution. Japanese Patent Publication No. 1-18855 discloses that a Bacillus subtilis-producing protease and a Aspergillus oryzae-producing protease are allowed to act on raw fish and shellfish in two steps at pH 6.0 to 7.0 and have a molecular weight of substantially 3,000. A method for producing a fish and shellfish extract having a pharmacological action by decomposing the following peptide amino acid groups and free amino acids and separating and concentrating the separated solution is disclosed in Japanese Patent Publication No. 5-73731. An immunostimulant containing, as an active ingredient, a proteolytic extract containing a peptide having a molecular weight of 3500 to 15000 obtained as described above has been proposed. In addition, a method has been proposed in which raw fish and shellfish are treated in one step with an acid-resistant protease to extract an effective extract, and a method in which the obtained extract is used for a seasoning, a growth agent and the like. ing.

[0003] Further, these conventional methods aim at decomposing proteins such as raw fish and shellfish to extract effective peptides and the like according to the intended use, and the purification step is, for example, centrifugation, vacuum separation, or the like. A method of separating an extract component decomposed by a protease from waste and a waste material is adopted.When the extract component is used as a drug or the like, an ion exchange membrane or the like is further added to the separated extract component. A high-purity active ingredient has been extracted by performing a purification step utilizing the same.

[0004] Recently, the influence of mineral components on humans or livestock has been emphasized. In particular, zinc components include alkaline phosphatase, alcohol dehydrokinase, decarboxylase, carboxypeptidase A, uricase, DNA polymerase, and RNA polymerase. In addition to acting as a component and active center of the zinc enzyme such as the above, it also has a function of activating the enzyme by forming an organic ligand, and has been proposed to play an important role. So now,
When the extract is used, for example, as a functional food, a feed additive for livestock, or the like, a new mineral component such as phosphorus, calcium, zinc or the like is added to adjust the component, and then the final product is produced.

On the other hand, it is known that various mineral components are contained in the raw fish and shellfish algae from which the extract is extracted. However, in the above-mentioned conventional method for extracting an effective extract, since the purpose is to extract an extract containing an effective peptide or the like according to the application, most of the mineral components contained in the raw material are contained in the separated waste. At present, it is discarded in the state where it was done.

[0006] Therefore, as a method of effectively utilizing the mineral components contained in the raw fish and shellfish algae, a step of purifying the mineral components is carried out before the step of separating the extract from the waste, It is possible to purify the mineral component from the extract and add it to the extract component.However, the addition of such a purification step leads to complication of the method, and furthermore, a new mineral component is added to the separated extract component in terms of cost. It has the disadvantage of being more expensive than current methods of addition and not suitable for industrial production.

[0007]

Accordingly, an object of the present invention is to extract an extract containing a large amount of a mineral component and an effective peptide component contained in fish and shellfish algae by an easy and simple method. It is an object of the present invention to provide a method for extracting a fish and shellfish algae extract containing a large amount of mineral components, which can be produced.

[0008]

According to the present invention, there is provided a means for solving] The raw fish shellfish algae proteolytically by proteolytic enzymes, a method of extracting the seafood algae extract was purified separated into the extract component and waste, autolysis 3. The pH at the time of proteolysis into raw fish and shellfish algae in which the enzyme has been deactivated is set to 4 . The present invention provides a method for extracting a fish and shellfish alga extract containing a large amount of a mineral component, characterized in that a mineral component is eluted in the extract by causing an acid-resistant proteinase to act when the value is less than 0.

Hereinafter, the present invention will be described in more detail. In the extraction method of the present invention, first, the raw fish and shellfish algae are decomposed by a protease, and the acid-resistant protease is decomposed at a specific pH.

As the raw fish and shellfish algae, for example, horse mackerel,
Mackerel, sardines, saury, skipjack, hockey, cod, squid,
Octopus, shrimp, oyster, clam, clam, mussel, mussel,
Red mussels, clams, seaweed, kelp, hijiki or mixtures thereof can be mentioned. The proteolytic enzyme is not particularly limited as long as it can decompose the protein, and can be variously selected according to the use of the obtained effective extract, the conditions of proteolysis, and the like. Specific examples include Bacillus subtilis-produced protease, Aspergillus-produced protease, and mixtures thereof. As described above, as the proteolytic enzyme, a known proteolytic enzyme which is conventionally used in a method in which a raw fish and shellfish algae is subjected to proteolysis with a protease and purified and separated into an extract component and waste to extract the fish and shellfish algal extract is used. Can be used but
When the protease is acted on in one step, it is necessary to use an acid-resistant protease. When the protease is acted on in multiple steps, it is necessary to use the acid-resistant protease at least once, preferably in the final step. Protease must be used. The use of the acid-resistant proteolytic enzyme is for decomposing the raw fish and shellfish algae protein to the molecular weight of the peptide according to various uses.

In order to decompose the raw fish and shellfish algae, for example, first, raw fish and shellfish algae, which are pre-treated such as shredding and slurrying, and a part of fish and shellfish from which a part of the internal organs of the fish and shellfish are taken out or a round. Pour the raw fish and shellfish algae into the reactor. Preferably, immediately after the introduction, the temperature is 75 ° C. or higher, particularly preferably 80 ° C.
It is desirable to raise the temperature to at least ℃ to deactivate the autolytic enzymes contained in the fish and shell algae, thereby removing odors such as fishy and shellfish algae's peculiar odor and odor. Then, at a temperature of 40 to 70 ° C., p
At an H of 5.0 to 7.0, preferably at a pH of 5.5 to 6.5, a protein contained in fish and shell algae is roughly degraded by adding an alkali-resistant protease. The crude degradation by the alkali-resistant protease is not always necessary, but is a step for relaxing the degradation conditions to the effective extract component by one stage of the acid-resistant protease. At this time, the decomposition time is not critical, but it is usually 30 minutes to 10 hours, preferably 1 to 6 hours.

When the crude decomposition is carried out with the alkali-resistant protease, the temperature should be at least 75 ° C.
Preferably, the temperature is raised to 80 ° C. or higher to completely inactivate the alkali-resistant protease. The inactivation time is usually from 10 minutes to 1 hour, preferably from 15 to 30 minutes. If this inactivation step is not carried out, the enzyme in the preceding step acts in the protein degradation step by acid-resistant protease, which is an essential step described later, and it is difficult to decompose the target effective peptide. Is not preferred.

The acid-degraded protease is added to the crude degraded product of the raw fish and shellfish algae or the alkali-resistant protease deactivating the autolytic enzyme, preferably at 30 to 60 ° C. to perform protein degradation. At this time, the pH should be less than 4.0.
You . pH 4 . In the case of 0 or more, the mineral components such as phosphorus, calcium, and zinc contained in the raw fish and shellfish algae and taurine do not elute into the extract containing the active peptide,
The mineral component content in the finally obtained fish and shell algae extract is extremely reduced. On the other hand, the lower limit of the pH is not particularly limited since the lower the pH, the more the amount of the mineral component eluted into the extract component is increased. However, when the treatment is performed at a high temperature from the viewpoint of operability and the like, the pH is 1.0 or more. Preferably 40 ° C
When the treatment is performed at the following low temperature, the pH is preferably 2.0 or more. The degradation time of the proteolysis by the acid-resistant protease is usually about 30 minutes to 5 hours, although it depends on the raw material components used for the degradation, the type of the acid-resistant protease, the molecular weight of the target effective peptide, and the like. be able to.

By performing the treatment with the acid-resistant protease under the above-mentioned specific pH conditions, the protein of the raw fish and shellfish algae is decomposed into an effective peptide component corresponding to the desired use, and the extract containing the effective peptide is contained in the extract. Then, a large amount of mineral components contained in the raw fish and shellfish algae elute. After the treatment with the acid-resistant protease, the temperature is preferably raised immediately to 75 ° C. or higher, particularly preferably 80 ° C. or higher, to completely inactivate the acid-resistant protease.

In the extraction method of the present invention, after the treatment with the acid-resistant proteolytic enzyme, the extract and waste are purified and separated to obtain the desired fish and shellfish algae extract containing a large amount of mineral components.

The purification and separation of the extract and the waste can be carried out by a conventional method according to, for example, a known vacuum filtration method, a centrifugation method or the like. The separated extract is an effective peptide depending on the desired use; free amino acids such as isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, and valine; taurine and potassium, phosphorus, magnesium, calcium, zinc,
Iron, molybdenum, aluminum, manganese, copper, lithium, tin, nickel, etc. It is a component with a water content of 90% by weight or more containing as a main component a mineral component corresponding to the raw fish and shellfish algae. And the like.

The obtained extract can be directly used as a target fish and shellfish algae extract containing a large amount of mineral components, and after the mineral components are eluted in the extract components,
A fish and shell algae extract containing a large amount of mineral components can be obtained by, for example, increasing the pH. The pH is
The pH is preferably raised to the original pH of the raw fish and shell algae, and more specifically, the pH is preferably adjusted to 5.5 to 7.0. Further, if necessary, organic minerals can be extracted using a purification method. Also, while maintaining the mineral content,
In order to further purify the effective peptides, purification using an ion-exchange resin membrane or the like can be performed according to a conventional method, and preferably, the water content is 30 to 6 at 60 ° C or less.
The target fish and shellfish algal extract can be obtained by a method such as concentration under reduced pressure to about 0%.

The fish and shellfish algae extract obtained by the extraction method of the present invention is a very safe substance without toxicity because it is composed entirely of natural products, and is a drug, various functional foods, drugs, various growth agents, The compounding amount and the like can be appropriately selected and used depending on the use of feed additives and seasonings for livestock. In addition, since many mineral components are contained, it is not necessary to add a new mineral component, which is conventionally performed, or the addition can be performed with a small amount of addition.

[0019]

According to the method for extracting algae extract of fish and shellfish of the present invention, the acid-resistant protease is allowed to act to change the pH in the conventional process such that the pH at the time of proteolysis is adjusted to a specific range. Mineral components contained in fish and shellfish algae can be eluted into extracts, so pharmaceuticals, various functional foods, drugs, various growth agents, livestock feed additives, It is possible to obtain a fish and shellfish algal extract containing an effective peptide and the like according to the use of a seasoning and the like, and also containing a large amount of mineral components contained in the raw fish and shellfish algae.

[0020]

The present invention will be described in more detail with reference to examples and comparative examples, but the present invention is not limited to these examples.

[0021]

[Reference Example 1] 2 kg of oysters are not pre-processed,
The mixture was placed in a reactor equipped with a stirrer together with 10 kg of water and heated to 80 ° C. After 15 minutes, the temperature is lowered to 50 ° C. and the pH is adjusted to 5.0.
2 g of a commercially available Bacillus subtilis-producing protease, trade name "Aloase AP-10" (manufactured by Yakult Biochemical KK)
Was added and reacted for 5 hours. Then the temperature was raised to 80 ° C,
After maintaining for 15 minutes, the mixture was cooled to 50 ° C. and adjusted to pH 4.0 by adding citric acid. To this was added 2 g of a koji mold-producing protease, trade name "Punchase NP-2" (manufactured by Yakult Biochemical KK), and the mixture was reacted for 3 hours. Thereafter, the temperature was raised to 80 ° C. to inactivate the enzyme again. This reaction solution was separated into an undecomposed layer such as an extract layer, an oil layer, and bone fragments by a centrifugal separator according to a conventional method.
Was concentrated under reduced pressure to prepare an oyster extract having a water content of 50% or less. Table 1 shows the proportions of the mineral component, the free amino acid component and the peptide amino acid group component. The content of the mineral component in the obtained extract component is indicated by the amount per 100 g of the extract, and the content of taurine, protein, free amino acid and peptide amino acid group is indicated by% by weight based on the total amount of the extract component.

[0022]

Example 1 pH at the time of adding Aspergillus-produced protease
Was changed to 2.0, and the oyster extract having a water content of 50% or less was prepared in the same manner as in Reference Example 1 except that the trade name "Punchase NP-2" was changed to the trade name "Sumiteam AP" (manufactured by Shin Nippon Chemical Industry Co., Ltd.). Prepared. Table 1 shows the proportions of the mineral component, the free amino acid component and the peptide amino acid group component.

[0023]

[Comparative Example 1] pH when adding Aspergillus-produced protease
Was changed to 6.0, and the same procedure as in Reference Example 1 was carried out.
Oyster extract of 0% or less was prepared. Table 1 shows the ratio of the mineral components, the free amino acid group and the peptide amino acid group component.
Shown in

[0024]

[Table 1]

[0025]

[Reference Example 2] except that the content of oyster and components used in Reference Example 1 using different oysters, was in to perform moisture below 50% oyster same manner as in Reference Example 1. Table 2 shows the proportions of the mineral components, the free amino acid group and the peptide amino acid group component.

[0026]

Example 2 pH at the time of adding Aspergillus-produced protease
Was changed to 3.0, the trade name “Punchase NP-2” to the trade name “Acid protease A” (manufactured by Amano Pharmaceutical Co., Ltd.), and the pH was set to 6.0 before concentration, in the same manner as in Reference Example 2. To give an oyster extract having a water content of 50% or less. Table 2 shows the ratio of the mineral component, the free amino acid component and the peptide amino acid group component.

[0027]

[Comparative Example 2] pH at the time of adding Aspergillus-produced protease
Was carried out in the same manner as in Reference Example 2 except that the oyster extract was adjusted to 5.0 to obtain an oyster extract having a water content of 50% or less. Table 2 shows the proportions of the mineral components, the free amino acid group and the peptide amino acid group component.

[0028]

[Table 2]

[0029]

[Reference Example 3] except that the content of oyster and components used in Reference Example 1 using different oysters, was in to perform moisture below 50% oyster same manner as in Reference Example 1. Table 3 shows the proportions of the mineral component, the free amino acid component, and the peptide amino acid group component.

[0030]

Example 3 pH at the time of adding Aspergillus-produced protease
Was changed to 2.5 and the trade name “Punchinase NP-2” was changed to “Purified Papain” (manufactured by Asahi Breweries).
An oyster extract having a water content of 50% or less was prepared in the same manner as in Reference Example 3 except that H was set to 7.0. Table 3 shows the proportions of the mineral components, the free amino acid group and the peptide amino acid group component.
Shown in

[0031]

Comparative Example 3 pH when adding Aspergillus-produced protease
Was carried out in the same manner as in Reference Example 3 except that the oyster extract was changed to 6.0 to obtain an oyster extract having a water content of 50% or less. Table 3 shows the proportions of the mineral component, the free amino acid component, and the peptide amino acid group component.

[0032]

[Table 3]

[0033]

REFERENCE EXAMPLE 4 2.5 kg of mackerel was placed in a reaction vessel equipped with a stirrer together with 12 kg of water as it was, without performing pretreatment.
Heated to ° C. After 15 minutes, lower the temperature to 50 ° C.
8.0, a commercially available Bacillus subtilis-producing protease, trade name "Aloase AP-10" (Yakult Biochemical KK.
2g) and reacted for 5 hours. Next, the temperature was raised to 80 ° C. and maintained for 15 minutes, then cooled to 50 ° C., and adjusted to pH 4.0 by adding acetic acid. In addition to this, a koji mold-producing protease, trade name "Punchase NP-2"
2 g (manufactured by Yakult Biochemicals KK) was added and reacted for 3 hours. Thereafter, the temperature was raised to 80 ° C. to inactivate the enzyme again. The reaction solution was separated into an extract layer, an oil layer, an undecomposed layer such as bone fragments by a centrifugal separator by a conventional method, and the extract layer was filtered and concentrated at 60 ° C. under reduced pressure to prepare a mackerel extract having a water content of 40% or less. Table 4 shows the proportions of the mineral components, the free amino acid group and the peptide amino acid group component.

[0034]

Comparative Example 4 pH at the time of adding Aspergillus-produced protease
Was changed to 6.0, and the same procedure as in Reference Example 4 was carried out to obtain a water content of 40%.
% Or less mackerel extract was prepared. Table 4 shows the proportions of the mineral components, the free amino acid group and the peptide amino acid group component.

[0035]

[Table 4]

Claims (2)

(57) [Claims] 1. A method for extracting a fish and shell algae extract by decomposing a raw fish and shell algae with a proteolytic enzyme, purifying and separating the extract and waste into an extract, and deactivating the autolytic enzyme.
3. The pH at the time of proteolysis into raw fish and shellfish algae is 4 . A method for extracting a fish and shellfish alga extract containing a large amount of a mineral component, wherein the mineral component is eluted in the extract component by allowing the acid-resistant protease to act as a value of less than 0. 2. The method for extracting a fish and shellfish alga extract containing a large amount of mineral components according to claim 1, wherein the pH is increased after the mineral components are eluted in the extract components.