JP6778094B2 - Production method of casein enzymatic degradation product and casein enzymatic degradation product - Google Patents

Description

The present invention relates to a method for producing a casein enzymatic degradation product and a casein enzymatic degradation product.

Conventionally, milk proteins have excellent nutritional properties in terms of in vivo usability and are widely used in the fields of foods, beverages and the like. Recently, the use of enzymatically degraded milk protein, which is obtained by enzymatically decomposing milk protein, is increasing for the purpose of improving stability under acidic conditions, improving digestion and absorption, reducing antigenicity, and the like.

However, when milk protein is hydrolyzed, there is a problem that unpleasant flavors such as bitterness are generated due to the generated tasting peptides and free amino acids, which has been a major restriction when using milk protein hydrolyzate. .. In addition, the generation of this unpleasant flavor is particularly remarkable due to the generation of bitterness when casein containing a large amount of hydrophobic amino acids is hydrolyzed. Therefore, when a casein enzymatically hydrolyzed product obtained by hydrolyzing casein with an enzyme is contained in food or drink and used, improvement of flavor has been an important issue.

Therefore, some techniques for improving the flavor have been developed. For example, Patent Document 1 discloses a casein enzymatic decomposition product that has almost no taste and odorless flavor and can be used for applications such as foods and beverages.

In addition, some milk protein enzyme decomposition products contain amino acids that are the source of umami, such as glutamic acid, and can express cheese-like flavor, umami, and richness. For example, Non-Patent Document 1 discloses a casein hydrolyzate which is a powder hydrolyzed using casein protein as a raw material by utilizing enzymatic decomposition and exhibits a strong umami taste and an aged cheddar cheese-like taste. ing.

However, with the prior art, it has been difficult to provide a casein enzymatic decomposition product having a strong umami taste while suppressing the bitterness of casein.

Japanese Unexamined Patent Publication No. 9-28306

Dairy Journal, 2016, Volume 6, pp.32-36

Therefore, the main purpose of this technique is to provide a casein enzymatic decomposition product having a strong umami taste and less bitterness.

As a result of diligent research to solve the above problems, the present inventors have conducted three types of enzymes: a proteolytic enzyme derived from a microorganism belonging to the genus Aspergillus, a proteolytic enzyme derived from lactic acid bacteria, and an enzyme having glutaminase activity. Surprisingly, it was found that a casein enzymatically decomposed product having a strong taste and less bitterness could be produced by enzymatically decomposing casein using the above-mentioned drug, and the present technology was completed.

That is, in the present technology, a proteolytic enzyme derived from a microorganism belonging to the genus Aspergillus, a proteolytic enzyme derived from lactic acid bacteria, and an enzyme having glutaminase activity are used for casein, and the content of free glutamate is 3800 mg. A method for producing a casein enzymatic degradation product in an amount of / 100 g or more , wherein the enzyme having glutaminase activity is a glutaminase derived from a microorganism belonging to the genus Bacillus .
In the production method according to the present technology, the enzyme having glutaminase activity can be added in an amount of 0.4% by mass or more with respect to casein.
Further, in the production method according to the present technology, the proteolytic enzyme derived from lactic acid bacteria can be added in an amount of 0.4% by mass or more with respect to casein. In this case, the content of the free glutamic acid can be 4400 mg / 100 g or more.
Further, in the production method according to the present technology, the casein enzymatic decomposition product can also be used as a flavor enhancer.

According to this technique, it is possible to provide a casein enzymatic decomposition product having strong umami and less bitterness. The effects described here are not necessarily limited, and may be any of the effects described in the present disclosure.

<1. Method for producing casein enzymatic degradation product>
The production method according to the present technology is characterized by using a proteolytic enzyme derived from a microorganism belonging to the genus Aspergillus, a proteolytic enzyme derived from lactic acid bacteria, and an enzyme having glutaminase activity with respect to casein, and the content of free glutamic acid is high. It is a method for producing a casein enzymatic degradation product having a weight of 3800 mg / 100 g or more.

(1) Casein In the production method according to this technology, casein, which is a raw material, is mainly composed of a protein derived from milk. The casein is not particularly limited, but a commercially available product may be used as the casein, or a refined product separated from milk may be used.
Examples of casein include various caseins, caseinates, rennet caseins, and the like, and these can be used alone or in combination of two or more.
Specifically, acid casein such as lactic acid casein, sulfuric acid casein, and hydrochloric acid casein; caseinate such as sodium caseinate, potassium caseinate, calcium caseinate, and magnesium caseinate, or a mixture of two or more of these. Can be mentioned.
In addition, bovine-derived casein or the like separated and purified from milk, skim milk, whole milk powder, whole fat powder milk, skim milk powder, or the like by a conventional method can also be used.

As described above, it has been difficult to provide a casein enzymatic decomposition product having a strong umami taste while suppressing the bitterness of casein by the prior art. In particular, it has been extremely difficult to produce a casein enzymatic degradation product having a free glutamic acid content of 3800 mg / 100 g or more.
However, the present inventors unexpectedly act on casein with three types of enzymes: a proteolytic enzyme derived from a microorganism belonging to the genus Aspergillus, a proteolytic enzyme derived from lactic acid bacteria, and an enzyme having glutaminase activity. It was also found that a casein enzymatic degradation product having a free glutaminase content of 3800 mg / 100 g or more can be obtained. Further, as confirmed by the present inventors, the casein enzymatic decomposition product also has a feature that the bitterness generated when casein is hydrolyzed is very small.

The casein enzymatic decomposition product produced by the production method according to this technology has high value by itself, but since it has the above-mentioned properties, it can be added to pharmaceuticals, foods and drinks, drinking feed, etc. to add flavor, richness, etc. It can be used as a flavor enhancer for enhancing the flavor.

Since casein used as a raw material is derived from milk, it has the advantages of being relatively inexpensive, stable, convenient, and available in large quantities. Therefore, in the production method according to the present technology using casein as a raw material, the casein enzymatic decomposition product can be obtained in a relatively inexpensive, safe, stable, convenient and large amount.

(2) Proteolytic enzyme derived from a microorganism belonging to the genus Aspergillus In the production method according to this technology, one or two proteolytic enzymes derived from a microorganism belonging to the genus Aspergillus that can be usually used in the field of pharmaceuticals or foods and drinks are used. More than a species can be freely selected and used.

Aspergillus (Aspergillus) genus includes, for example, Aspergillus oryzae, Aspergillus glaucus, Aspergillus sojae, Aspergillus tamarii, Aspergillus tamarii, Aspergillus tamarii, Aspergillus tamarii awamori) etc. can be mentioned.

Among these, it is preferable to use a proteolytic enzyme derived from Aspergillus oryzae in the present technology.

In this technique, the amount of proteolytic enzyme derived from a microorganism belonging to the genus Aspergillus is not particularly limited, and depends on the desired yield of the casein enzyme degradation product to be produced, the amount of two other types of enzymes to be added in combination, and the like. , Can be set as appropriate. In the present technology, it is particularly preferable to add 0.1% by mass or more, more preferably 0.5% by mass or more, and further preferably 2% by mass or more with respect to casein. This makes it possible to efficiently and stably produce a casein enzymatic decomposition product having a high content of free glutamic acid.

Further, in the present technology, from the viewpoint of cost and prevention of complication of the manufacturing process, it is preferable to add 20% by mass or less of the proteolytic enzyme derived from a microorganism belonging to the genus Aspergillus to casein, and add 10% by mass or less. It is more preferable, and it is further preferable to add 5% by mass or less.

(3) Lactic Acid Bacteria-Derived Proteolytic Enzymes In the production method according to this technology, one or more types of lactic acid bacteria-derived proteolytic enzymes that can be usually used in the field of pharmaceuticals or foods and drinks are freely selected and used. be able to. The "lactic acid bacterium" is a general term for bacteria that produce lactic acid by metabolism, and examples thereof include Lactococcus bacteria, Lactobacillus bacteria, Streptococcus bacteria, and Bifidobacterium bacteria. Bifidobacterium bacteria can also be called "Bifidobacterium", which is a general term for bacteria that produce lactic acid and acetic acid by metabolism.

The proteolytic enzyme derived from lactic acid bacteria can be produced, for example, by the method described in the section "(3) Enzyme to be used" in column 6, line 4 of Japanese Patent Publication No. 54-36235.
Specifically, lactic acid bacteria (including "Bifidobacterium") are cultured by a known method (for example, the method described in Japanese Patent Publication No. 48-43878), and the obtained culture solution is centrifuged to separate lactic acid bacteria cells. , Suspend the cells in sterile water, centrifuge and collect the lactic acid bacteria cells twice, wash the cells, suspend the cells in sterile water at a concentration of 20%, and collect the bacteria. The cells are crushed by a body crusher (for example, manufactured by Willy Bachnfen Engineering; KDL type) and lyophilized to obtain a powder of a proteolytic enzyme derived from lactic acid bacteria.

Examples of bacteria belonging to the genus Lactococcus include Lactococcus lactis, L. lactis subsp. Lactis, and Lactococcus lactis subsp. Lactis. Bacteria such as L. lactis subsp. Lactis biovar. Diacetylactis and L. lactis subsp. Cremoris can be mentioned.

Examples of Lactobacillus genus bacteria include Lactobacillus delbrueckii subsp. Lactis and L. delbrueckii subsp. Bulgaricus. , Lactobacillus bulgaricus (L. bulgaricus), Lactobacillus reuteri (L. reuteri), Lactobacillus herlveticus (L. helveticus) and the like.

Examples of the Streptococcus genus bacterium include bacteria such as Streptococcus salivarius subsp. Thermophilus and S. salivarius subsp. Thermophilus.

Bifidobacterium genus bacteria include, for example, Bifidobacterium longum, B. breve, Bifidobacterium bifidum, and Bifidobacterium. Examples include bacteria such as Bifidobacterium longum infantis (B. infantis) and Bifidobacterium longum (B. animalis).

In the present technology, among these, it is preferable to use a proteolytic enzyme derived from a bacterium belonging to the genus Lactobacillus, and it is more preferable to use a proteolytic enzyme derived from L. helveticus.

In this technique, the amount of the proteolytic enzyme derived from lactic acid bacteria is not particularly limited, and is appropriately set according to the target yield of the casein enzyme-degraded product to be produced, the amount of the other two types of enzymes to be added in combination, and the like. can do. In the present technology, it is particularly preferable to add 0.4% by mass or more, more preferably 0.6% by mass or more, and further preferably 0.8% by mass or more with respect to casein. As a result, a casein enzymatic degradation product having a high content of free glutamic acid can be efficiently and stably produced, and in particular, a casein enzymatic degradation product having a free glutamic acid content of 4400 mg / 100 g or more is stable. It becomes easy to manufacture.

Further, in the present technology, from the viewpoint of cost and prevention of complication of the manufacturing process, the proteolytic enzyme derived from lactic acid bacteria is preferably added in an amount of 20% by mass or less, more preferably 10% by mass or less, based on casein. It is more preferable to add 5% by mass or less.

(4) Enzyme having glutaminase activity In the production method according to this technology, one or more kinds of enzymes having glutaminase activity that can be usually used in the field of pharmaceuticals or foods and drinks can be freely selected and used. it can. The "glutaminase activity" is an enzymatic activity that catalyzes a hydrolysis reaction that produces L-glutamic acid using free L-glutamine as a substrate.

Examples of the enzyme having glutaminase activity include glutaminase derived from a microorganism belonging to the genus Bacillus, glutaminase derived from a microorganism belonging to the genus Aspergillus, and the like. In this technique, the enzyme having glutaminase activity does not have to be an enzyme having only glutaminase activity, and may be an enzyme having both a rearrangement reaction and an asparaginase reaction.

Microorganisms belonging to the genus Bacillus include, for example, Bacillus subtilis, Bacillus amyloliquefaciens, Bacillus coagulans, Bacillus lentus, Bacillus lentus, and Bacillus lentus.・ Examples include Bacillus licheniformis. Further, as a glutaminase derived from a microorganism belonging to the genus Bacillus, a glutaminase produced by a modified bacterium such as gene recombination by applying biotechnology may also be used.

Examples of microorganisms belonging to the genus Aspergillus include Aspergillus oryzae, Aspergillus soyer, Aspergillus niger and the like. Further, as a glutaminase derived from a microorganism belonging to the genus Aspergillus, a glutaminase produced by a modified bacterium such as genetic recombination by applying biotechnology may also be used.

Among these, in the present technique, it is particularly preferable to use glutaminase derived from a microorganism belonging to the genus Bacillus, and it is more preferable to use glutaminase derived from Bacillus subtilis.

In the present technology, the amount of the enzyme having glutaminase activity is not particularly limited, and is appropriately set according to the desired yield of the casein enzyme degradation product to be produced, the amount of the other two types of enzymes to be added in combination, and the like. be able to. In the present technology, it is particularly preferable to add 0.4% by mass or more, and more preferably 0.8% by mass or more with respect to casein. This makes it possible to efficiently and stably produce a casein enzymatic decomposition product having a high content of free glutamic acid.

Further, in the present technology, from the viewpoint of cost and prevention of complication of the manufacturing process, it is preferable to add 10% by mass or less of the enzyme having glutaminase activity to casein, and more preferably 5% by mass or less. It is more preferable to add 1% by mass or less.

(5) Other Enzymes In the manufacturing method according to this technology, one or more types of other enzymes that can be usually used in the field of pharmaceuticals or foods and drinks can be freely used as long as the effects of this technology are not impaired. It can be selected and used. Further, in the present technology, the amount of other enzymes added is not particularly limited, and is appropriately set according to the desired yield of the casein enzymatic decomposition product to be produced, the amount of the other three types of enzymes to be added in combination, and the like. be able to.

(6) Specific Example of Production Method The production method according to this technique is free from casein using a proteolytic enzyme derived from a microorganism belonging to the genus Aspergillus, a proteolytic enzyme derived from lactic acid bacteria, and an enzyme having glutaminase activity. The detailed steps are not particularly limited as long as a casein enzymatic degradation product having a glutamic acid content of 3800 mg / 100 g or more can be obtained.
In the production method according to the present technology, various steps used in the known production method of casein enzymatic decomposition products can be freely selected and adopted.
Hereinafter, an example of the manufacturing method will be specifically described.

[Preparation of raw material solution]
A casein-containing solution is prepared by dissolving or dispersing a raw material containing casein in a solvent such as water.

Casein used as a raw material is as described above.
Examples of raw materials other than casein include pH adjusters, surfactants, and the like, and one or more of them can be selected.
The solvent is not particularly limited, but it is preferable to use distilled water, RO water (water treated with a reverse osmosis membrane), well water or the like as water.
The casein concentration in the casein-containing solution is not particularly limited, but is usually preferably in the concentration range of about 5 to 15% by mass in terms of protein from the viewpoint of efficiency and operability.

The pH of the casein-containing solution is preferably pH 4 to 10, more preferably pH 6 to 10, further preferably pH 6.2 to 9, and preferably pH 6.5 to 9. Especially preferable.
Inorganic acids such as hydrochloric acid and sulfuric acid; organic acids such as acetic acid can be used for pH acid adjustment, and alkali metal salts such as sodium hydroxide; alkaline soil such as calcium hydroxide can be used for pH alkali adjustment. Kind metal salts and the like can be used. In the present technology, these may be used alone or in combination of two or more.

[Enzyme reaction]
Next, the above-mentioned three types of enzymes (proteolytic enzyme derived from a microorganism belonging to the genus Aspergillus, proteolytic enzyme derived from lactic acid bacteria, and enzyme having glutaminase activity) are added to the casein-containing solution. In the present technology, the order of addition of these enzymes is not particularly limited.

Further, in the present technology, the progress state of the enzyme treatment can be appropriately adjusted by adjusting the reaction conditions such as the reaction duration, the reaction temperature, and the initial pH.
For example, by monitoring the enzymatic reaction, the duration of the reaction can be determined so that the physicochemical properties of the casein enzymatic degradation product are at the desired values.
Examples of the method for monitoring the enzyme reaction include a method in which a part of the reaction solution is collected and the decomposition rate of the protein is measured. The reaction duration may be determined by measuring the collected reaction solution as it is.

In the present technology, the lower limit of the reaction time is preferably 3 hours or more, more preferably 5 hours or more, still more preferably 10 hours or more, and particularly preferably 15 hours or more. The upper limit is preferably 24 hours or less, more preferably 20 hours or less.
The reaction time is preferably 3 to 24 hours, more preferably 5 to 24 hours, still more preferably 10 to 20 hours, and particularly preferably 15 to 20 hours.

In the present technique, the reaction pH is preferably pH 6-9, more preferably 6.2-9. A more preferable upper limit value is 9. Further, a more preferable lower limit value is pH 6.3. The reaction pH is preferably pH 6 or higher in order to suppress acid precipitation of casein.

In the present technology, the lower limit of the reaction temperature is preferably 20 ° C. or higher, more preferably 40 ° C. or higher, and further preferably 50 ° C. or higher. The upper limit is preferably 65 ° C. or lower, more preferably 60 ° C. or lower, and even more preferably 55 ° C. or lower.
The reaction temperature is preferably 20 to 65 ° C, more preferably 40 to 60 ° C, still more preferably 50 to 60 ° C, and particularly preferably 50 to 55 ° C. The reaction temperature is preferably 60 ° C. or lower in order to suppress the heating inactivation of the enzyme.

Next, the enzymatic reaction is stopped.
The enzyme reaction is stopped by inactivating the enzyme in the hydrolyzate. The deactivation treatment can be carried out by a conventional method, for example, a heat deactivation treatment or the like.
The conditions for the heat deactivation treatment (heating temperature, heating time, etc.) can be appropriately set in consideration of the thermal stability of the enzyme used.
In the present technology, as an example, the enzyme can be inactivated by holding at 80 to 85 ° C. for 5 to 10 minutes.

[Separation / purification]
The enzymatic reaction product obtained by carrying out the above-mentioned enzymatic reaction may be separated / purified by a conventional separation / purification method. Thereby, impurities may be removed from the enzymatic decomposition product according to the present technology. Further, the enzymatic decomposition product according to the present technology may be in a solution state. Further, in the present technology, a desired fraction or a desired peptide may be recovered from the enzymatic decomposition product according to the present technology.

Examples of the separation / purification method include a filtration method, a microfiltration method, a membrane separation treatment method such as a limit filtration membrane, a resin adsorption separation method, a column chromatography method, an HPLC separation / purification method, a precipitation method, and a crystal method. .. In the present technology, these can be used alone or in combination of two or more.

The filtration method can be carried out by a known method, for example, using diatomaceous earth and using a known device.

The membrane separation treatment method can be carried out using a known device. The known apparatus is not particularly limited, but for example, an ultrafiltration module Pellicon2 Cassettes (manufactured by Merck Millipore, molecular weight cut-off 1,000 Da), an ultrafiltration module SEP1053 (manufactured by Asahi Kasei Co., Ltd., molecular weight cut-off). 3,000 Da), SIP1053 (manufactured by Asahi Kasei Co., Ltd., molecular weight cut-off 6,000 Da), SLP1053 (manufactured by Asahi Kasei Co., Ltd., molecular weight cut-off 10,000 Da) and the like.

The resin adsorption separation method can be carried out by a known method, for example, by bringing an enzymatic decomposition product into contact with a resin. The resin is not particularly limited, and examples thereof include an ion exchange resin, a chelate resin, an affinity adsorption resin, a synthetic adsorbent, and a resin for high performance liquid chromatography. Specific examples thereof include trade names: Diaion, Sepabeads (manufactured by Mitsubishi Chemical Corporation), Amberlite XAD (manufactured by Organo Corporation), KS-35 (manufactured by Ajinomoto Fine Techno Co., Ltd.) and the like.
The resin adsorption separation method can also be carried out by a continuous method in which these resins are filled in a column and the enzymatic decomposition products are continuously flowed in and out, and the resin is put into the enzymatic decomposition products. Then, after contacting with each other for a certain period of time, the enzymatic decomposition product and the resin can be separated by a batch method.

The precipitation method can be carried out by a known method, and examples thereof include an acid precipitation method and a centrifugation method. In the present technology, these can be used alone or in combination of two or more.

[Sterilization]
In addition, in this technique, the obtained casein enzymatic decomposition product may be sterilized. As the sterilization method, a conventional heat treatment method or the like can be used.
The heating temperature and holding time during the heat treatment may be appropriately set under conditions that allow sufficient sterilization. For example, sterilization can be performed by heat treatment at 70 to 140 ° C. for 2 seconds to 30 minutes.
The heat sterilization method can be either a batch method or a continuous method, and even in the continuous method, a plate heat exchange method, an infusion method, an injection method, or the like can be used.

[Concentration treatment / drying treatment / powdering treatment / secondary treatment]
Further, the obtained casein enzymatic decomposition product can be used as it is in a solution state, and if necessary, the solution can be used as a concentrated concentrated solution by a known method. Further, the concentrated solution can be dried by a known method and used as a powder.

<2. Casein Enzymatic Degradation>
The present technology also provides a casein enzymatic degradation product having a free glutamic acid content of 3800 mg / 100 g or more. Since the casein enzymatic decomposition product according to this technology contains a high content of free glutamic acid known as an umami component, it is added to various pharmaceuticals, foods and drinks, drinking feeds, etc. to enhance the flavor such as umami and richness. It can be used as a flavor enhancer. In addition, the casein enzymatic decomposition product according to the present technology is highly useful because it has very little bitterness, as shown in Examples described later.

The method for producing the casein enzymatic decomposition product according to the present technology is not particularly limited, but it is desirable to produce it by the above-mentioned production method according to the present technology. The casein enzymatic degradation product produced by the production method according to the present technology is excellent in safety because it is produced using casein as a raw material and usually using three types of enzymes that can be used in the pharmaceutical or food fields. Even if it is taken continuously for a long period of time, there is little need to worry about side effects, which is very useful.

The content of free glutamic acid of the casein enzymatic decomposition product according to the present technology is not particularly limited as long as it is 3800 mg / 100 g or more, but is preferably 4000 mg / 100 g or more, more preferably 4200 mg / 100 g or more, and 4400 mg / 100 g or more. It is more preferably 100 g or more, and particularly preferably 4800 mg / 100 g or more.

In addition, the casein enzymatic decomposition product according to the present technology also has an effect of making the salty taste strongly felt as shown in Test Examples 2 and 3 described later, and therefore, it is also used as a flavor enhancer for salt reduction. Can be done.

When the casein enzymatic degradation product according to the present technology is used for a pharmaceutical product, it can be prepared by adding the casein enzymatic degradation product according to the present technology to a known pharmaceutical product, or the casein enzymatic degradation product is mixed with the raw material of the pharmaceutical product. It is also possible to manufacture new medicines.
Further, when the casein enzymatic decomposition product according to the present technology is used as a pharmaceutical product, it may be used as it is, after being concentrated, or after being processed into a solid, granular or powder form.

The drug can be appropriately formulated into a desired dosage form according to an administration method such as oral administration or parenteral administration. The dosage form is not particularly limited, but in the case of oral administration, for example, solid preparations such as powders, granules, tablets, troches, capsules; preparations in liquid preparations such as solutions, syrups, suspensions and emulsions. Can be transformed into. In the case of parenteral administration, it can be formulated into, for example, suppositories, sprays, inhalants, ointments, patches, injections and the like. In the present technology, it is preferable to formulate the dosage form for oral administration.
The formulation can be carried out by a known method as appropriate according to the dosage form.

At the time of formulation, a formulation carrier may be appropriately added to the formulation. Further, in addition to the casein enzymatic decomposition product according to the present technology, components such as excipients, pH adjusters, colorants, and flavoring agents usually used for formulation can be used. Further, a component having a known or future-finding effect of preventing, treating and / or ameliorating a disease or symptom can be appropriately used in combination depending on the purpose.

As the preparation carrier, various organic or inorganic carriers can be used depending on the dosage form.
Examples of the carrier in the case of a solid preparation include excipients, binders, disintegrants, lubricants, stabilizers, flavoring agents and the like.

Examples of excipients include sugar derivatives such as lactose, sucrose, glucose, mannit, and sorbit; starch derivatives such as corn starch, horse bell starch, α-starch, dextrin, and carboxymethyl starch; crystalline cellulose, hydroxypropyl cellulose, etc. Cellulose derivatives such as hydroxypropylmethyl cellulose, carboxymethyl cellulose, carboxymethyl cellulose calcium; gum arabic; dextran; purulan; silicate derivatives such as light anhydrous silicic acid, synthetic aluminum silicate, magnesium aluminometasilicate; phosphate derivatives such as calcium phosphate; carbonic acid Carbonated carbonate derivatives such as calcium; sulfate derivatives such as calcium sulfate can be mentioned.

Examples of the binder include gelatin; polyvinylpyrrolidone; macrogol, etc., in addition to the above-mentioned excipients.

Examples of the disintegrant include, in addition to the above-mentioned excipients, chemically modified starch or cellulose derivatives such as croscarmellose sodium, carboxymethyl starch sodium, and crosslinked polyvinylpyrrolidone.

Examples of the lubricant include talc; stearic acid; metal stearic acid salts such as calcium stearate and magnesium sulfate; colloidal silica; waxes such as pea gum and gay wax; boric acid; glycol; carboxylic acids such as fumaric acid and adipic acid. Sodium carboxylic acid salts such as sodium benzoate; sulfates such as sodium sulfate; leucine; lauryl sulfates such as sodium lauryl sulfate and magnesium lauryl sulfate; silicates such as silicic acid anhydride and silicate hydrate; starch derivatives and the like. Be done.

Examples of the stabilizer include paraoxybenzoic acid esters such as methylparaben and propylparaben; alcohols such as chlorobutanol, benzyl alcohol and phenylethyl alcohol; benzalconium chloride; acetic acid anhydride; sorbic acid and the like.

Examples of the flavoring and flavoring agent include sweeteners, acidulants, and flavors.
Examples of the carrier used in the case of a liquid preparation for oral administration include a solvent such as water, a flavoring and odorant, and the like.

The content of the casein enzymatic decomposition product according to the present technology in the above-mentioned pharmaceutical products is not particularly limited, but it is desirable that the content is within a range in which the flavor improving effect can be efficiently obtained and can be ingested reasonably.

Further, in the present technology, when the casein enzymatic decomposition product according to the present technology is used for foods and drinks for humans or animals, it can be prepared by adding the casein enzymatic decomposition product according to the present technology to known foods and drinks. It is also possible to produce a new food or drink by mixing the casein enzymatic decomposition product with the raw material of the food or drink.
Further, when the casein enzymatic decomposition product according to the present technology is used for foods and drinks, it may be used as it is, after being concentrated, or after being processed into a solid form, a granular form or a powder form.

Foods and drinks may be liquid, pasty, solid, powder, etc., in addition to tablets, liquid foods, etc., for example, wheat flour products, instant foods, processed agricultural products, processed marine products, processed livestock products, milk. -Dairy products, fats and oils, basic seasonings, complex seasonings / foods, frozen foods, confectionery, beverages, and other commercial foods.

Examples of dairy products include fermented milk, dairy beverages, lactic acid bacteria beverages, sweetened milk powder, skim milk powder, sweetened milk powder, adjusted milk powder, cream, cheese, butter, ice cream and the like.
Examples of wheat flour products include bread crumbs, macaroni, spaghetti, noodles, cake mixes, fried chicken flour, bread crumbs and the like.
Examples of instant foods include instant noodles, cup noodles, retort / cooked foods, canned foods, microwave foods, instant soups / stews, instant miso soup / soups, canned soups, freeze / dried foods, and other instant foods. Be done.
Examples of processed agricultural products include canned agricultural products, canned fruits, jams and marmalades, pickles, boiled beans, dried agricultural products, cereals (processed grain products), and the like.
Examples of processed marine products include canned marine products, fish hams and sausages, fish paste products, marine delicacies, and boiled fish.
Examples of processed livestock products include canned livestock and pastes, livestock ham and sausage.
Examples of fats and oils include butter, margarines, vegetable oils and the like.
Examples of the basic seasonings include soy sauce, miso, sauces, processed tomato seasonings, mirins, vinegars and the like, and the compound seasonings / foods include cooking mixes, curry ingredients, sauces, etc. Examples include dressings, mentsuyu, spices, and other complex seasonings.
Examples of frozen foods include raw material frozen foods, semi-cooked frozen foods, and cooked frozen foods.
Examples of confectioneries include caramel, candy, chewing gum, chocolate, cookies, biscuits, cakes, pies, snacks, crackers, Japanese confectionery, rice confectionery, bean confectionery, dessert confectionery, and other confectionery.
Beverages include, for example, carbonated beverages, natural fruit juices, fruit juice beverages, refreshing beverages containing fruit juice, fruit meat beverages, fruit beverages containing fruit grains, vegetable beverages, soymilk, soymilk beverages, coffee beverages, tea beverages, powdered beverages, concentrated beverages. , Sports beverages, nutritional beverages, alcoholic beverages, other favorite beverages and the like.
Examples of commercial foods other than the above-mentioned foods include baby food, sprinkle, tea mashed seaweed and the like.

In addition, the food and drink defined in the present technology can be provided and sold as food and drink for which insurance purposes (for example, for salt reduction, etc.) are displayed.

In the present technology, the "display" act includes all acts for informing the consumer of the said use, and if the expression can remind or analogize the said use, the purpose and display of the display. Regardless of the content, the object to be displayed, the medium, etc., all of them fall under the "display" act of the present technology.

Further, it is preferable that the "display" is performed by an expression that allows the consumer to directly recognize the application. Specifically, the act of transferring a product related to food and drink or the packaging of the product with the above-mentioned use, displaying it for delivery, transfer or delivery, and importing it, advertising on the product, price list or transaction documents Examples include the act of describing the above-mentioned use and displaying or distributing it, or describing the above-mentioned use in the information containing these and providing it by an electromagnetic (Internet, etc.) method.

On the other hand, as the display content, it is preferable that the display is approved by the government or the like (for example, a display obtained based on various systems established by the government and performed in a manner based on such approval). In addition, it is preferable to attach such display contents to packaging, containers, catalogs, pamphlets, advertising materials at sales sites such as POP (Point of purchase advertising), and other documents.

In addition, "labeled" includes health foods, functional foods, foods for the sick, enteral nutrition foods, special purpose foods, health functional foods, specified health foods, functional labeled foods, nutritionally functional foods, and pharmaceutical departments. Labeling as a foreign product can also be mentioned. Among these, in particular, labeling approved by the Consumer Affairs Agency, for example, a food system for specified health use, a food system with functional labeling, and labeling approved by a system similar to these can be mentioned. More specifically, labeling as food for specified health use, labeling as conditional food for specified health use, labeling as functional food, labeling indicating that it affects the structure and function of the body, labeling for reducing disease risk, etc. Can be mentioned. Among these, as a typical example, labeling as a food for specified health use stipulated in the Health Promotion Law Enforcement Regulations (April 30, 2003, Ministry of Health, Labor and Welfare Ordinance No. 86) (especially labeling for health uses) ), Labeling as functionally labeled foods stipulated in the Food Labeling Law (Act No. 70 of 2013) and labeling similar to these.

It should be noted that the wording used for displaying as described above is not limited to the wording such as "for salt reduction", and any other wording may be a wording indicating the salt reduction effect or the like. Needless to say, it is included in the scope of the present technology. As such a wording, for example, it is possible to display based on various uses such that the consumer is made to recognize the salt reduction effect and the like.

Further, in the present technology, when the casein enzymatic decomposition product according to the present technology is used for drinking feed, it can be prepared by adding the casein enzymatic decomposition product according to the present technology to a known drinking feed, or as a raw material for drinking feed. A new drinking feed can also be produced by mixing the casein enzymatic decomposition product therein.
Further, when the casein enzymatic decomposition product according to the present technology is used for drinking feed, it may be used as it is, after being concentrated, or after being processed into a solid form, a granular form or a powder form.

As raw materials for drinking feed, for example, cereals such as corn, wheat, barley, and rye; bran such as bran, wheat bran, rice bran, and defatted rice bran; production lees such as corn gluten meal and corn jam meal; defatted milk powder, Animal feeds such as whey, fish flour, and bone meal; yeasts such as beer yeast; mineral feeds such as calcium phosphate and calcium carbonate; fats and oils; amino acids; sugars and the like. In addition, examples of the form of the drinking feed include pet food drinking feed (pet food and the like), livestock feeding feed, fish feeding feeding feed and the like.

As described above, the casein enzymatic decomposition product according to the present technology can be used in a wide range of fields such as pharmaceuticals, foods and drinks, and drinking feed.

Further, in this technology, it is also possible to adopt the following configuration.
[1] Use of a proteolytic enzyme derived from a microorganism belonging to the genus Aspergillus, a proteolytic enzyme derived from lactic acid bacteria, and an enzyme having glutaminase activity for producing a casein enzymatic degradation product having a free glutamic acid content of 3800 mg / 100 g or more. ..
[2] The use according to [1], wherein 0.4% by mass or more of the enzyme having glutaminase activity is added to casein.
[3] The use according to [1] or [2], wherein 0.4% by mass or more of the proteolytic enzyme derived from lactic acid bacteria is added to casein.
[4] The use according to [3], wherein the content of the free glutamic acid is 4400 mg / 100 g or more.
[5] Use of a proteolytic enzyme derived from a microorganism belonging to the genus Aspergillus, a proteolytic enzyme derived from lactic acid bacteria, and an enzyme having glutaminase activity for producing a casein enzymatic degradation product having a free glutamic acid content of 4400 mg / 100 g or more. ..
[6] The use according to any one of [1] to [5], wherein the casein enzymatic decomposition product is used as a flavor enhancer.
[7] Use of a casein enzymatic degradation product having a free glutamic acid content of 3800 mg / 100 g or more as a flavor enhancer.
[8] The use according to [7], wherein the flavor enhancer is used for reducing salt.

Hereinafter, the present technology will be described in more detail based on Examples. It should be noted that the examples described below show examples of typical examples of the present technology, and the scope of the present technology is not narrowly interpreted by this.

[Manufacturing example]
<Manufacturing example 1>
Commercially available casein (protein content 90%, manufactured by Fonterra) was swollen in purified water at a concentration of 10%, and then a 10% sodium hydroxide solution was added and dissolved. Next, the temperature was adjusted to 50 ° C., and at each addition amount shown in Table 1 below, "Proteax (derived from Aspergillus oryzae; manufactured by Amano Enzyme)" and "FD-H (derived from Lactobacillus helveticus; manufactured by Morinaga Dairy Co., Ltd.)" , And "Glutaminase SD-C100S (derived from Bacillus subtilis; manufactured by Amano Enzyme)" were added and treated for 16 hours, and then the enzyme was inactivated by heating at 90 ° C. for 10 minutes, as shown in Table 1 below. The casein enzymatic degradation products of Examples 1 to 13 were produced. Each casein enzymatic degradation product was dried by a conventional method to obtain a powdered casein enzymatic degradation product.

[Test example]
<Test Example 1>
The content (mg / 100 g) of free glutamic acid was measured for each casein enzymatic decomposition product or each casein hydrolyzate shown in Tables 1 and 2 below. In addition, the flavor (umami and bitterness) was evaluated by 10 panelists. Specifically, 10 points were evaluated as the strongest or the best, and the average value of the 10-grade evaluation by each panelist was calculated and used as each evaluation point. The numerical values in Tables 1 and 2 indicate this evaluation point.

In Examples 1 to 13, each casein enzymatic decomposition product produced in Production Example 1 described above was used.
For Comparative Example 1, the casein hydrolyzate HCP337 manufactured by Tatsua Japan Co., Ltd. described in Dairy Journal, 2016, Volume 6, pp.32-36 was used. As Comparative Example 2, the casein hydrolyzate of Example 1 described in JP-A-9-28306 was used. In Comparative Examples 3 to 14, each enzyme was added at each addition amount shown in Table 2 below, and each casein enzymatic decomposition product produced by the same method as in Production Example 1 described above was used.

Further, as shown in Tables 1 and 2, "Papain (derived from porcine pancreas; manufactured by Amano Enzyme)" was used in Examples 12 and 13 and Comparative Examples 11 and 12 as enzymes other than the above-mentioned three types. In Comparative Example 13, "Papain W-40 (derived from papain; manufactured by Amano Enzyme)" was used, and in Comparative Example 14, "Bioplase SP-20 (derived from Bacillus sp.; manufactured by Nagase Chemtech)" was used.

As shown in Tables 1 and 2 above, casein is allowed to act on casein by three types of enzymes: a proteolytic enzyme derived from a microorganism belonging to the genus Aspergillus, a proteolytic enzyme derived from lactic acid bacteria, and an enzyme having glutaminase activity. It was found that a casein enzymatic degradation product having a high content of free glutamate (= 3800 mg / 100 g or more) and less bitterness can be obtained.

In Example 10, the content of free glutamine was 233 mg / 100 g, and the total content of free glutamic acid and free glutamine was 5805 mg / 100 g. Further, in Example 13, the content of free glutamine was 118 mg / 100 g, and the total content of free glutamic acid and free glutamine was 5878 mg / 100 g.
On the other hand, in Comparative Example 6, the content of free glutamine was 1336 mg / 100 g, and the total content of free glutamic acid and free glutamine was 4059 mg / 100 g.

Example 13 differs from Example 10 in that 1.0% by mass of pancreatin is added, but even if pancreatine is added, there is a large difference in the total content of free glutamic acid and free glutamine. Therefore, it is considered essential to use the above-mentioned three types of enzymes in this technique.

Further, Example 10 is different from Comparative Example 6 in that 0.8% by mass of glutaminase is added, but there is a large difference in the total contents of free glutamic acid and free glutamine (= 1746 mg / 100 g). Was recognized. This suggests that the production of glutamine, which is a substrate, may not be simply due to two types of enzymes, a proteolytic enzyme derived from a microorganism belonging to the genus Aspergillus and a proteolytic enzyme derived from lactic acid bacteria. Specifically, in the absence of glutaminase, some of the N-terminal glutamine and free glutamine in the peptide are easily converted to pyroglutamic acid, but on the other hand, an enzyme having glutaminase activity should be used. Since the above-mentioned glutamine can be rapidly converted to stable glutamic acid, it is expected that a caseine enzymatic decomposition product having a high content of glutamic acid is obtained.

<Test Example 2>
24 g of Hokkaido sweet corn powder (manufactured by Natural Kitchen Co., Ltd. 10) and 1 g of salt were dissolved in 125 ml of hot water to prepare corn soup. In the corn soup, among the casein enzymatic decomposition products used in Test Example 1, the casein enzymatic decomposition products or each casein hydrolyzate shown in Table 3 below were added to a concentration of 1.0% (w / v). It was added and dissolved, and the flavor (richness, umami, saltiness and bitterness) was evaluated by 10 panelists. Since the method for evaluating the flavor is the same as that in Test Example 1, the description thereof is omitted here. The numerical values in Table 3 indicate the above-mentioned evaluation points.

As shown in Table 3 above, when the casein enzymatic decomposition product of Example 10 was used, there was a rich feeling, the richness was the strongest, and the salty taste was also strong. In addition, the bitterness peculiar to casein enzymatic decomposition products was not felt. On the other hand, the additive-free product had a weak flavor as a whole. Further, when the casein hydrolyzate of Comparative Example 1 was used, the bitterness peculiar to the casein hydrolyzate was slightly felt, although the richness was felt as compared with the additive-free product. Further, when the casein hydrolyzate of Comparative Example 2 was used, the bitterness peculiar to the casein hydrolyzate was strongly felt, and the overall flavor was also weak as compared with Example 10.

<Test Example 3>
After frying 1/4 onion and 1 teaspoon of garlic tube in olive oil, add 1/2 can of cut tomato and bay leaf and fry on medium heat, add 0.2 g of salt to adjust the tomato sauce. In the tomato sauce, 0.5% (w / v) of each casein enzymatic decomposition product or each casein hydrolyzate shown in Table 4 below was added to the casein enzymatic decomposition product used in Test Example 1, or 1. The enzyme was added and dissolved to a concentration of 0% (w / v), and the flavor (richness, umami, saltiness and bitterness) was evaluated by 10 panelists. Since the method for evaluating the flavor is the same as that in Test Example 1, the description thereof is omitted here. The numerical values in Table 4 indicate the above-mentioned evaluation points.

As shown in Table 4 above, when 0.5% (w / v) of the casein enzymatic decomposition product of Example 10 was used, the aftertaste was moderately rich and umami, and the overall taste was thickened. , Salty taste was also felt. Further, when 1.0% (w / v) of the casein enzymatic decomposition product of Example 10 was used, umami was felt from the first taste, the overall flavor was strong, there was a rich feeling, and the salty taste was also strongly felt. .. On the other hand, the additive-free product had a weak flavor as a whole. Further, when 1.0% (w / v) of the casein hydrolyzate of Comparative Example 1 was used, although the body was richer than that of the additive-free product, the bitterness peculiar to the casein hydrolyzate was felt. The overall flavor was inferior to 10. Further, when 1.0% (w / v) of the casein hydrolyzate of Comparative Example 2 was used, the bitterness peculiar to the casein hydrolyzate was strongly felt, and the overall flavor was compared with that of Example 10. Was also weak.

Claims (5)

Casein having a free glutamic acid content of 3800 mg / 100 g or more, which comprises using a proteolytic enzyme derived from a microorganism belonging to the genus Aspergillus, a proteolytic enzyme derived from lactic acid bacteria, and an enzyme having glutaminase activity with respect to casein. It is a method for producing enzymatic degradation products .
The method for producing a casein enzymatic degradation product, wherein the enzyme having glutaminase activity is a glutaminase derived from a microorganism belonging to the genus Bacillus. The method for producing a casein enzymatic degradation product according to claim 1, wherein 0.4% by mass or more of the enzyme having glutaminase activity is added to casein. The method for producing a casein enzymatic degradation product according to claim 1 or 2, wherein 0.4% by mass or more of the proteolytic enzyme derived from lactic acid bacteria is added to casein. The method for producing a casein enzymatic degradation product according to claim 3, wherein the content of the free glutamic acid is 4400 mg / 100 g or more. The method for producing a casein enzymatic degradation product according to any one of claims 1 to 4, wherein the casein enzymatic degradation product is used as a flavor enhancer.