JPH0819385A - Method for extracting essence of fish, shellfish and alga containing large quantity of mineral component - Google Patents

Abstract

PURPOSE:To provide an industrially usable and simple method for obtaining essences of fishes, shellfishes or algae containing effective peptides, etc., corresponding to uses such as medicines, various functional foods, drugs, various growth agents, feed additives for domestic animals, seasonings, etc., and further a large quantity of mineral components included in the resources. CONSTITUTION:This method for extracting essences of fishes, shellfishes or algae having a large quantity of mineral components is to carry out proteolysis of fishes, shellfishes or algae by a protease, purify and separate an extract and waste. The proteolysis is carried out by applying an acid resistant protease at <pH5.0. to elute the mineral components into the essence. By this method the mineral components included in fishes, shellfishes and algae resources can be eluted into the essence solely by changing pH within a specific range during the usual proteolysis.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION The present invention relates to pharmaceuticals, functional foods,
It is useful for medicines, various growth agents, feed additives for livestock, seasonings, etc., and minerals such as phosphorus, calcium, zinc, iron, potassium contained in raw material fish and shellfish algae, and a large amount of taurine-rich minerals. The present invention relates to a method for extracting contained fish and shellfish algae extracts.

[0002]

2. Description of the Related Art Conventionally, various methods have been proposed in which a proteolytic enzyme is allowed to act on fish and shellfish algae to separate it into an extract and wastes to extract the fish and shellfish algae extract. 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. Specifically, for example, JP-B-53-31935 discloses that the temperature is 50 to 60 ° C., p
In H9-10, a method of adding an alkali-resistant protease to the raw fish to decompose the protein, adjusting the pH to 5-6, adding / decomposing the acid-resistant protease, and concentrating the separated liquid Japanese Patent Publication No. 14885/1989 discloses that raw fish and shellfish are allowed to act in a two-step manner with a Bacillus subtilis-produced protease and an Aspergillus-produced protease at a molecular weight of 3000. The 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 liquid is disclosed in Japanese Examined Patent Publication No. 5-73731 and decomposes fish with a proteolytic enzyme. An immunostimulant containing a proteolytic extract containing the thus obtained peptide having a molecular weight of 3500 to 15000 as an active ingredient has been proposed. In addition, a method of treating raw fish and shellfish with an acid-resistant proteolytic enzyme in one step to extract an effective extract, a method of using the obtained extract for a seasoning, a growth agent, etc. are proposed. ing.

Further, these conventional methods are aimed 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 thereof is, for example, a centrifugal separation method or a vacuum separation method. A method of separating the extract component decomposed by a proteolytic enzyme and the wastes is adopted, and when the extract component is used as a drug or the like, an ion exchange membrane or the like is added to the separated extract component. The active ingredient with high purity is extracted by carrying out a purification process utilizing

By the way, recently, the influence of mineral components on humans or livestock has been emphasized. Particularly, zinc components are alkaline phosphatase, alcohol dehydrokinase, decarboxylase, carboxypeptidase A, uricase, DNA polymerase, RNA polymerase. It has been proposed that it plays an important role in that it functions as a constituent component of zinc enzymes such as the above, as an active center, and also functions as forming an organic ligand to activate the enzyme. So now,
When the extract is used, for example, as a functional food or a feed additive for livestock, it is finalized after adding new mineral components such as phosphorus, calcium and zinc to adjust the components.

On the other hand, it is known that various mineral components are contained in the raw fish, shellfish and 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 purpose, most of the mineral components contained in the raw material are contained in the separated waste. The current situation is that they are discarded in the state of being discarded.

Therefore, as a method of effectively utilizing the mineral components contained in the raw material fish and shellfish algae, a purification process of the mineral components is carried out before the separation process of the above-mentioned extract and the waste products, or the separated waste products. It is possible to purify the mineral component from the extract and add it to the extract, but the addition of such a purification step complicates the method, and a new mineral component is added to the separated extract in terms of cost. It has the disadvantage of being more expensive than the current method of addition and unsuitable for industrial production.

[0007]

Therefore, an object of the present invention is to extract an extract containing a large amount of a mineral component contained in fish and shellfish algae and an effective peptide component by an easy and simple method. An object of the present invention is to provide a method of extracting a fish and shellfish alga extract containing a large amount of mineral components.

[0008]

According to the present invention, a method of proteolytically decomposing raw material fish and shellfish algae with a proteolytic enzyme and purifying and separating into extract and waste to extract a fish and shellfish algae extract Provided is a method for extracting a large amount of mineral component-containing fish and shellfish alga extract, which is characterized in that a pH during proteolysis by acting a proteolytic enzyme is set to less than 5.0 and a mineral component is eluted in the extract. .

The present invention will be described in more detail below. In the extraction method of the present invention, the raw material fish and shellfish algae are first subjected to protein degradation with a protease, and at this time, the degradation with an acid-resistant protease is carried out at a specific pH.

Examples of the raw material fish and shellfish algae include horse mackerel,
Mackerel, sardines, saury, bonito, hockey, cod, squid,
Octopus, shrimp, oyster, blue clam, clam, mussel, mussel,
Examples thereof include red mussel, clam, seaweed, kelp, hijiki or a mixture thereof. The proteolytic enzyme is not particularly limited as long as it can decompose a protein, and various kinds can be selected depending on the use of the obtained effective extract, the conditions for proteolysis and the like. Specific examples thereof include Bacillus subtilis-produced protease, Aspergillus-produced protease, or a mixture thereof. As described above, as the proteolytic enzyme, a known one used in the method of extracting a fish and shell alga extract by proteolytically decomposing the raw material fish and shell alga with a proteolytic enzyme and purifying and separating it into an extract and a waste product is used. Can be used,
When the proteolytic enzyme acts in one step, it is necessary to use an acid-resistant proteolytic enzyme, and when the proteolytic enzyme acts in multiple steps, the acid-resistant protease should be used at least once, preferably in the final step. It is necessary to use proteolytic enzymes. The use of the acid-resistant proteolytic enzyme is for degrading the protein of the raw material fish and shellfish alga to the molecular weight of the peptide according to various uses.

In order to proteolytically decompose the above-mentioned raw fish and shellfish algae, for example, the raw fish and shellfish algae which have been subjected to pretreatment such as shredding and slurrying, part of the internal organs of the fish and shellfish, or the portion of the fish shellfish or the whole Put whole fish and shellfish algae into the reaction can. Immediately after the addition, the temperature is preferably 75 ° C. or higher, particularly preferably 80.
It is desirable to raise the temperature to ℃ or more to deactivate the autolytic enzyme contained in the fish and shellfish algae to remove the raw shavings, malodors and the like peculiar to the fish and shellfish algae. Then, the temperature is 40 to 70 ° C., p
At H5.0 to 7.0, preferably pH 5.5 to 6.5, an alkali-resistant protease is added to roughly decompose proteins contained in fish and shellfish algae. It is not always necessary to perform the crude decomposition with the alkali-resistant protease, but it is a step for relaxing the decomposition conditions up to the effective extract component by one step of the acid-resistant protease. At this time, the decomposition time is not critical, but is usually 30 minutes to 10 hours, preferably 1 to 6 hours.

In the case of performing the crude decomposition with the alkali-resistant protease, the temperature is at least 75 ° C. or higher,
It is preferable to raise the temperature to 80 ° C. or higher to completely inactivate the alkali-resistant protease. The inactivation time is usually 10 minutes to 1 hour, preferably 15 to 30 minutes. If this inactivation step is not carried out, in the proteolytic step by an acid-resistant proteolytic enzyme, which is an essential step to be described later, the enzyme in the previous step acts together to make it difficult to decompose into the target effective peptide. Is not desirable.

The acid-degrading protease is added to the crude fish-and-sea alga in which the autolytic enzyme is inactivated or the crude decomposed product by the alkali-resistant protease, preferably at 30 to 60 ° C., for protein degradation. At this time, the pH is less than 5.0, preferably 4.5 or less, and particularly preferably less than 4.0. When the pH is 5.0 or higher, the mineral components such as phosphorus, calcium, zinc and taurine contained in the raw material shellfish algae do not elute on the side of the extract containing the effective peptide, and the fish finally obtained. The content of mineral components in shellfish algae extract is extremely reduced. On the other hand, the lower limit of pH is not particularly limited because the lower the pH, the more the elution amount of the mineral component into the extract increases, but from the viewpoint of operability, the pH is 1.0 or more. Is preferred and 4
When treating at a low temperature of 0 ° C. or lower, pH of 2.0 or higher is preferable. The proteolytic degradation time by the acid-resistant protease is usually about 30 minutes to 5 hours, although it varies depending on the raw material components to be used for the decomposition, the type of acid-resistant protease, the molecular weight of the target effective peptide, and the like. be able to.

By performing treatment with an acid-resistant proteolytic enzyme under the above-mentioned specific pH conditions, the protein of the raw material fish and shellfish algae is decomposed into effective peptide components according to the desired use, and the extract containing the effective peptide is decomposed. In addition, a large amount of mineral components contained in the raw fish, shellfish and algae are eluted. After the treatment with the acid-resistant protease, the temperature is preferably raised 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 component and the waste product are purified and separated to obtain the desired algae extract containing a large amount of mineral components.

Purification and separation of the extract and the waste can be carried out by a conventional method, for example, according to a known vacuum filtration method, centrifugal separation method or the like. The separated extract is an effective peptide according to the desired application; isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, free amino acids such as valine; taurine and potassium, phosphorus, magnesium, calcium, zinc,
Iron, molybdenum, aluminum, manganese, copper, lithium, tin, nickel, etc. A raw material is a component having a water content of 90% by weight or more, which contains a mineral component according to the raw fish, shellfish and algae as the main component, and the waste is an oil layer and bone fragments. It is an undecomposed material such as.

The extract thus obtained can be used as it is as a target fish and shellfish alga extract containing a large amount of mineral components, and after the mineral components are eluted into the extract,
The target fish and shellfish alga extract containing a large amount of mineral components can be obtained by a method of increasing pH and the like. The pH is
It is preferable to raise the pH to the original pH of the raw fish, shellfish and algae, and specifically, it is desirable to adjust the pH to 5.5 to 7.0. If necessary, the organic mineral can be extracted by using a purification method. In addition, while maintaining the content of mineral components,
Further, in order to highly purify the effective peptides, purification using an ion exchange resin membrane or the like can be carried out by a conventional method, and preferably, the water content is 30 to 6 at 60 ° C. or lower.
The target fish and shell alga extract can be obtained by a method such as vacuum concentration to about 0%.

The fish and shellfish alga extract obtained by the extraction method of the present invention is a non-toxic and extremely safe substance because it consists entirely of natural products, and is a drug, various functional foods, drugs, various growth agents, The blending amount and the like of the livestock feed additive, seasoning and the like can be appropriately selected and used. Further, since a large amount of mineral component is contained, it is not necessary to add a new mineral component which is conventionally used, or it can be carried out by adding a small amount.

[0019]

INDUSTRIAL APPLICABILITY In the method for extracting a fish and shellfish alga extract of the present invention, the acid-resistant proteolytic enzyme is allowed to act, and the pH in the case of proteolysis is changed to a specific range by simply changing the condition in the conventional process. Since the mineral components contained in fish and shellfish algae can be eluted into the extract, it is a simple method that can be used industrially, such as pharmaceuticals, various functional foods, drugs, various growth agents, feed additives for livestock, It is possible to obtain a fish and shellfish alga extract containing an effective peptide or the like depending on the use such as a seasoning and also containing a large amount of mineral components contained in the raw fish and shellfish alga.

[0020]

EXAMPLES The present invention will be described in more detail with reference to Examples and Comparative Examples, but the present invention is not limited thereto.

[0021]

[Example 1] 2 kg of oysters were not pretreated and were left as they were,
It was put in a reaction can equipped with a stirrer together with 10 kg of water and heated to 80 ° C. After 15 minutes, lower the temperature to 50 ° C and adjust to pH 5.0.
Bacillus subtilis-produced proteolytic enzyme, trade name "Aroase AP-10" (manufactured by Yakult Biochemical KK) in Japan
Was added and reacted for 5 hours. Then raise the temperature to 80 ° C,
After maintaining for 15 minutes, the mixture was cooled to 50 ° C., and citric acid was added to adjust the pH to 4.0. 2 g of a koji mold-produced proteolytic enzyme, trade name "Punchtase NP-2" (manufactured by Yakult Biochemical KK) was added to this and reacted for 3 hours. Then, the temperature was raised to 80 ° C. to inactivate the enzyme again. This reaction solution is separated into an extract layer, an oil layer, an undecomposed layer such as bone fragments by a centrifuge by a conventional method, and the extract layer is filtered at 60 ° C.
Was concentrated under reduced pressure to prepare an oyster extract having a water content of 50% or less. Table 1 shows the ratios 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 shown by the amount per 100 g of the extract, and the contents of taurine, protein, free amino acid and peptide amino acid group are shown by weight% with respect to the total amount of the extract component.

[0022]

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

[0023]

[Comparative Example 1] pH at the time of adding the protease of Aspergillus oryzae
The same procedure as in Example 1 was repeated, except that the water content was changed to 6.0.
An oyster extract of 0% or less was prepared. Table 1 shows the proportions of mineral components, free amino acids and peptide amino acids.
Shown in

[0024]

[Table 1]

[0025]

Example 3 An oyster extract having a water content of 50% or less was prepared in the same manner as in Example 1 except that the oysters used in Example 1 were different from each other in the content of each component. Table 2 shows the proportions of the mineral component, the free amino acid group and the peptide amino acid group component.

[0026]

[Example 4] pH at the time of adding a protease produced by Aspergillus oryzae
In the same manner as in Example 3, except that the trade name "Punchtase NP-2" was changed to "Acid protease A" (manufactured by Amano Pharmaceutical Co., Ltd.) and the pH was adjusted to 6.0 before concentration. The oyster extract having a water content of 50% or less was obtained. Table 2 shows the ratios 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 a protease produced by Aspergillus oryzae
The oyster extract having a water content of 50% or less was obtained in the same manner as in Example 3 except that the oyster extract was changed to 5.0. Table 2 shows the proportions of the mineral component, the free amino acid group and the peptide amino acid group component.

[0028]

[Table 2]

[0029]

Example 5 An oyster extract having a water content of 50% or less was prepared in the same manner as in Example 1 except that the oysters used in Example 1 had different contents of each component. Table 3 shows the ratios of the mineral component, the free amino acid component and the peptide amino acid group component.

[0030]

[Example 6] pH at the time of adding Aspergillus oryzae-produced protease
2.5, the trade name "Punchatase NP-2" is changed to the trade name "Purified papain" (manufactured by Asahi Breweries), and after concentration p
An oyster extract having a water content of 50% or less was obtained in the same manner as in Example 4 except that H was 7.0. Table 3 shows the proportions of mineral components, free amino acids and peptide amino acids.
Shown in

[0031]

[Comparative Example 3] pH at the time of adding the protease of Aspergillus oryzae
Was carried out in the same manner as in Example 4 except that the oyster extract had a water content of 50% or less. Table 3 shows the ratios of the mineral component, the free amino acid component and the peptide amino acid group component.

[0032]

[Table 3]

[0033]

[Example 7] Mackerel 2.5 kg was put into a reaction can equipped with a stirrer together with 12 kg of water without pretreatment,
Heated to ° C. After 15 minutes, lower the temperature to 50 ° C and adjust the pH.
Commercially available Bacillus subtilis proteolytic enzyme in 8.0, trade name "Aroase AP-10" (Yakult Biochemistry KK
2 g), and reacted for 5 hours. Then, the temperature was raised to 80 ° C. and maintained for 15 minutes, then cooled to 50 ° C. and acetic acid was added to adjust the pH to 4.0. Proteolytic enzyme produced by Aspergillus oryzae, product name "Punchase NP-2"
(Yakult Biochemical KK) 2 g was added and reacted for 3 hours. Then, the temperature was raised to 80 ° C. to inactivate the enzyme again. This reaction solution was separated into an extract layer, an oil layer, an undegraded layer such as bone fragments by a centrifugal separator by a conventional method, and the extract layer was filtered and concentrated under reduced pressure at 60 ° C. to prepare a mackerel extract having a water content of 40% or less. Table 4 shows the proportions of the mineral component, the free amino acid group and the peptide amino acid group component.

[0034]

[Comparative Example 4] pH at the time of adding Aspergillus oryzae proteolytic enzyme
Was performed in the same manner as in Example 6 except that the water content was changed to 6.0.
% Mackerel extract was prepared. Table 4 shows the proportions of the mineral component, the free amino acid group and the peptide amino acid group component.

[0035]

[Table 4]

Claims (2)

[Claims] 1. A method of proteolytically degrading a raw fish and shellfish algae with a proteolytic enzyme to purify and separate into an extract and a waste product to extract a fish and shellfish algae extract, by acting an acid-resistant proteolytic enzyme. A method for extracting a fish and shellfish alga extract containing a large amount of mineral components, characterized in that the pH at the time is less than 5.0 and the mineral components are eluted into the extract. 2. The method for extracting a seaweed alga extract containing a large amount of mineral components according to claim 1, wherein the pH is raised after the mineral components are eluted in the extract.