JP4468831B2 - Fermented acerola product containing a large amount of γ-aminobutyric acid and method for producing the same - Google Patents

Description

  The present invention relates to an acerola food enriched with γ-aminobutyric acid and a method for producing the same.

  Acerola is a tropical fruit belonging to the genus Hollytranoaceae and is native to the Caribbean Islands. Acerola fruit contains about 1,500 mg / 100 g or more of vitamin C, and has recently been known as a plant containing abundant vitamin C, and is now used as a drink and health food in various countries around the world.

  Vitamin C is one of the important nutritional components indispensable for maintaining the life of human beings because it has important functions such as antioxidant action, immunity enhancing action, and whitening action. Vitamin C is also involved in bone metabolism, osteoblast differentiation and proliferation, and collagen production and maintenance. Collagen is the main component of connective tissue such as skin, bone, and tendon, and is contained in almost all tissues and organs. When vitamin C is deficient, collagen production decreases, blood vessels become brittle, arteriosclerosis occurs, and blood pressure rises. Collagen also constitutes the membrane that wraps the brain membrane, so if vitamin C is insufficient and collagen cannot be produced sufficiently, the function of the brain membrane is dulled, and the function of nerve cells in the brain is also reduced. Memory, thinking, judgment, etc. become dull and so-called blurred symptoms occur. Therefore, vitamin C is one of the very important nutrients for human beings.

  For the above reasons, acerola juice is favored by people as a health drink, and its production and sales are increasing year by year. At the same time, research on acerola juice has been extensively conducted, and many related technologies have been applied for patents (Patent Documents 1 to 4). Many of the techniques described in these documents focus on the functionality of components contained in the acerola juice itself, particularly vitamin C, and the functionality that is hardly contained in the acerola juice itself due to fermentation reactions and the like. There are few techniques for producing many components and increasing the added value of acerola juice.

  On the other hand, recently, γ-aminobutyric acid (γ-aminobutyric acid, hereinafter sometimes abbreviated as GABA) has been recognized as a functional component by people and has received wide attention. GABA is a kind of amino acid having a non-protein composition widely distributed in nature, and is contained in various mushrooms, fruits, vegetables, cereals and the like as a component of food. In vivo, there are many in the substantia nigra in the brain, basal ganglia and the like. GABA has many physiological effects such as dilating blood vessels to lower blood pressure, activating blood flow in the brain, increasing oxygen supply to brain cells, and promoting enhancement of metabolic function and memory. The effect is mentioned. Focusing on this preferable action, techniques for generating GABA in various food materials have been disclosed so far (Patent Documents 5 to 9). Unfortunately, the acerola juice does not contain much GABA component, and a method for increasing the GABA content of the acerola juice has not been developed yet.

JP-A-5-207865 JP-A-5-344846 JP-A-6-022727 JP-A-11-206341 Japanese Patent Laid-Open No. 9-238650 Japanese Patent Laid-Open No. 10-295394 JP-A-11-103825 JP 2000-210075 A JP 2003-70462 A

  As mentioned above, both vitamin C and GABA are important functional ingredients for the living body, and some complementary actions are seen between these two ingredients, and they are taken together as food ingredients. If possible, it can be expected that each functionality can be demonstrated and synergistic effects can be expected. However, under the present circumstances, foods containing a high amount of vitamin C and foods containing a high content of GABA have been developed and sold, respectively, but a food containing a large amount of vitamin C and GABA simultaneously has not been developed yet. People have to eat a variety of food products at the same time to consume these important ingredients. It goes without saying that this is a problem both in terms of cost and effort. If these two ingredients are provided in large amounts in one food, it is considered that there is no doubt that they can greatly contribute to people's healthy life. That is, a technique for further increasing the added value of acerola juice by increasing the GABA content of acerola juice is one of the important research subjects.

  The present inventors noticed that acerola juice contains a large amount of saccharides such as glucose and fructose in addition to containing a lot of vitamin C, and as a result of earnest research to solve the above-mentioned problems, the acerola juice contains GABA. Addition of yeast having high production ability, particularly yeast of Pichia or Candida, and fermenting can produce a large amount of GABA without adding other additives, and acerola juice by fermentation Since the content of vitamin C occupying the solid content is increased by consuming a large amount of excess carbohydrate contained therein, it has been found that low-sugar and highly functional acerola juice can be obtained. It came to be completed.

The present invention includes the following inventions.
(1) Fermentation containing γ-aminobutyric acid by allowing yeast having the ability to produce γ-aminobutyric acid by fermentation reaction in the presence of acerola fruit or processed product thereof or a processed product thereof to acerola fruit or processed product thereof A method of manufacturing an acerola product.
(2) The method according to (1), wherein the yeast is a yeast belonging to the genus Pichia or Candida.
(3) The method according to (1) or (2), wherein the yeast is Pichia anomala MR-1 (Accession No. FERM BP-10134) or a mutant strain having the ability to produce γ-aminobutyric acid.
(4) The method according to any one of (1) to (3), wherein the fermentation reaction is performed under conditions of an initial pH of 3.0 to 5.0 and a temperature of 32 to 45 ° C.
(5) The method according to any one of (1) to (4), wherein the fermentation reaction is performed in the presence of glutamic acid or a salt thereof.
(6) The method according to any one of (1) to (5), wherein the fermentation reaction is performed in a nitrogen atmosphere.
(7) Fermented acerola product containing γ-aminobutyric acid produced by the method according to any one of (1) to (6).
(8) The fermented acerola product containing γ-aminobutyric acid according to (7), which is in the form of a beverage, powder, tablet or jelly.
(9) A food containing 50 mg or more of γ-aminobutyric acid per 100 g of solids and 500 mg or more of vitamin C per 100 g of solids.

  According to the present invention, fermented acerola products enriched with GABA, foods and drinks using the products, and methods for producing them are provided.

  Hereinafter, the present invention will be described in more detail. In the present specification, “%” means “% by weight” unless otherwise specified.

  In the present invention, the acerola fruit refers to the entire fruit including the edible part and the seed core part. As long as the acerola fruit contains vitamin C and sugars, the production area and variety are not particularly limited.

  In the present invention, the processed product of acerola fruit refers to an acerola fruit processed by a normal processing method. The processed product of acerola fruit is not particularly limited as long as vitamin C and saccharides contained in the fruit are contained, and examples thereof include acerola puree, acerola fruit juice, and acerola fruit crushed material. Acerola puree refers to those obtained by crushing acerola fruit by a mechanical method and removing seed nuclei contained therein. Acerola juice is usually obtained by filtering and purifying acerola puree. In view of production costs and the like, it is preferable to use acerola puree as a raw material for fermentation. The acerola fruit crushed material refers to a product obtained by crushing and pulverizing the entire acerola fruit or the edible portion with a mixer or the like.

  The acerola fruit or the processed product thereof can be appropriately subjected to a treatment such as an extraction treatment or lyophilization.

  When acerola puree is used as a raw material in the present invention, it can be used as it is without any pretreatment in advance, but in order to cause the reaction to proceed more quickly, these acerola puree raw materials are added with sterilized water. It is more preferable to use after diluting 3 times. Moreover, since the pH of the raw material at the time of dilution is in the range of about 3.0 to 4.0 and is suitable for the fermentation reaction by yeast, there is no need to bother adjusting the pH of the reaction solution.

  In the GABA production reaction in the present invention, saccharide is used as a reaction substrate. Since the acerola fruit or the processed product thereof contains saccharides (mainly monosaccharides such as glucose and fructose) (for example, acerola puree contains about 2-3% of saccharides), the present invention Since the GABA production reaction in is carried out in the presence of sugars, it is not necessary to add further sugars to the acerola fruit or its processed product (Test Example 1). Therefore, the method of the present invention has a very simple procedure and enables mass production of a GABA-containing fermented acerola product at a lower cost.

  If only a predetermined yeast is added to the acerola fruit or its processed product, a large amount of GABA component can be produced without adding other reaction accelerators, etc., but in order to further increase the amount of GABA produced It is preferable to add glutamic acid or a salt thereof (for example, glutamic acid sodium salt) to the reaction raw material. The amount of glutamic acid or a salt thereof added is not particularly limited. For example, as shown in Test Example 3, for a reaction mixture containing 50% acerola puree and 10% predetermined yeast (water content of about 80%), A concentration in the range of about 0.4 to 0.8% is preferred. Even if the amount of glutamic acid added is further increased, not only a further increase in the amount of GABA produced can be expected, but excessive glutamic acid or a salt thereof remains in the GABA-containing fermented acerola product, which adversely affects the acerola flavor. Sometimes.

  Fermentation acerola containing γ-aminobutyric acid by allowing yeast having the ability to produce γ-aminobutyric acid by fermentation reaction in the presence of the acerola fruit or the processed product thereof or the processed product thereof to the acerola fruit or the processed product thereof You can get a product. Examples of yeast having such ability include yeasts belonging to the genus Saccharomyces such as baker's yeast, brewer's yeast, sake yeast, and the like, as well as yeasts belonging to the genus Pichia or Candida, and yeast belonging to the genus Pichia or Candida. Is particularly preferred. More specifically, Pichia anomala (for example, Pichia anomala MR-1 (Accession No. FERM BP-10134, dated September 28, 2004) Deposited), Pichia anomala NBRC-10213, Pichia anomala NBRC-100267), Pichia jadinii, anamorph: Candida utilis (eg Pichia jazini NBRC-0987), Candida utilis (Candida il) -Uchiris NBRC-10707), but is not limited to this. Pichia anomala MR-1 is particularly preferred. As long as it has the ability to produce γ-aminobutyric acid, mutants of Pichia anomala MR-1 are also preferably used. Mutagenesis treatment can be performed using any suitable mutagen. Here, the term “mutagen” should be understood in a broad sense to include not only a drug having a mutagenic effect but also a treatment having a mutagenic effect such as UV irradiation. Examples of suitable mutagens include ethyl methanesulfonate, UV irradiation, N-methyl-N'-nitro-N-nitrosoguanidine, nucleotide base analogs such as bromouracil, and acridines, but any other effect A typical mutagen can also be used. Any of these yeasts can be used in the method of the present invention as a suspension of yeast cells themselves. These yeasts can also be used in the form of so-called immobilized yeast supported on an appropriate carrier. Examples of the “processed product” of yeast in the present invention include this immobilized yeast.

  Although experimental data are not shown in this specification, the GABA production reaction by Pichia anomala MR-1 has the following characteristics. (1) As long as saccharides or sugar metabolism intermediates are present, a large amount of GABA can be produced even under conditions where glutamic acid or a salt thereof hardly exists. (2) Not only is GABA produced in large amounts in the presence of saccharides or sugar metabolism intermediates, but other components such as free alanine are also produced. (3) A large amount of GABA is hardly observed even when glutamic acid or a salt thereof is added under the condition where no saccharide or sugar metabolism intermediate exists. (4) A large amount of ethanol component is detected in the GABA production reaction solution in the presence of saccharides or sugar metabolism intermediates. (5) When the cells are stored frozen for 2 days or more and then used for the GABA production reaction, the amount of GABA produced is greatly reduced by about 50% or more than that of cells refrigerated for the same number of days.

  Assuming that GABA production by Pichia anomala MR-1 is based on a simple enzymatic reaction by a specific enzyme, such as GABA production in lactic acid bacteria, whether the cell is a dead cell or a living cell As long as the related enzyme is not inactivated, there should be no significant difference in the amount of GABA produced, but this does not explain the phenomenon (5) above. Therefore, GABA production by Pichia anomala MR-1 is presumed to be due to a kind of fermentation that occurs in combination with metabolic functions in the cells. This is supported by the simultaneous production of Ala and ethanol as described in (2) and (4) above. Therefore, when GABA is produced by the method of the present invention, it is expected that other useful components can also be produced simultaneously. In addition, GABA production by Pichia anomala MR-1 is advantageous in that it does not require a preliminary treatment such as acetone treatment such as normal yeast and can be used as it is. As shown in Test Example 6, the ability of Pichia or Candida yeast to produce GABA by viable cells is extremely high as compared to other yeasts. GABA production by other yeast strains preferably used in the present invention, such as Pichia anomala NBRC-10213, Pichia anomala NBRC-100267, Pichia jazini NBRC-0987, Candida utilis NBRC-10707, is also similarly fermented. It is estimated that

  About the addition density | concentration of the yeast used for this invention, as shown in Test Example 1 and Example 2 (Pichia Anomala MR-1 yeast) which will be described later, relative to the total amount of the reaction liquid, 80%) is sufficient if it is in the range of about 4 to 15% by weight. Even if the amount of yeast added is further increased, a significant increase in the amount of GABA produced cannot be expected so much. On the contrary, a significant increase in the amount of yeast added may cause a noticeable yeast odor and impair the flavor unique to acerola fruit. is there.

  In the method of the present invention, the initial pH in the reaction solution is preferably 3.0 to 5.0, more preferably 3.0 to 4.0.

  The reaction temperature in the fermentation reaction with the yeast of the present invention is not particularly limited as long as it is a condition that can produce a desired amount of GABA, but is preferably in the range of 32-53 ° C, more preferably in the range of 32-45 ° C.

  In the fermentation reaction of the present invention, it has been found that a larger amount of GABA is produced under conditions where yeast cells are difficult to grow, that is, under conditions where the yeast is actually in a growth-arrested state. Yeast cell growth is temperature dependent. For example, under the reaction conditions of the present invention, growth is suppressed and GABA production is increased under “high temperature” conditions of 40 ° C. or higher (particularly 40 ° C. or higher and 45 ° C. or lower). Growth is observed and the amount of GABA produced is suppressed. Therefore, from the viewpoint of effectively suppressing cell growth and increasing the amount of GABA production, a temperature condition of 40 ° C. or higher, particularly 40 to 45 ° C. is preferable.

  Note that, under “high temperature” conditions of 40 ° C. or higher, useful components such as functional components such as vitamin C, carotenoids, and polyphenols are oxidized and easily broken. On the other hand, at a “low temperature” of 40 ° C. or lower, the yeast proliferates remarkably using the organic components in the acerola fruit as nutrients, and it may not be possible to obtain the expected high GABA production. When there is such a problem, it is possible to appropriately fill the reaction mixture with nitrogen before the start and perform the fermentation reaction under a constant nitrogen atmosphere to suppress yeast growth and oxidation of useful components. The degree of nitrogen filling is not particularly limited, but it is preferably such that the fermentation reaction by yeast is not suppressed. In consideration of various conditions such as the degree of fermentation reaction, temperature, and time, when charged with nitrogen, the temperature ranges from 32 to 40 ° C. for 24 to 48 hours, and when not charged with nitrogen, 40 to 45 ° C. Reaction within 12 to 24 hours within the temperature range is preferred.

  The present invention also relates to a GABA-containing fermented acerola product obtained by the above method. The GABA-containing fermented acerola product includes a fermentation reaction product obtained by the above method or a processed product thereof. The treated product as used herein refers to, for example, a deactivation treatment by a method such as heating, a product subjected to a normal separation and purification treatment such as centrifugation and filtration, a product which is appropriately concentrated to a desired concentration by vacuum concentration or the like, or It refers to powdered by freeze drying, spray drying, etc.

  The fermented acerola product obtained by the present invention has abundant vitamin C originally contained in the acerola fruit, and at the same time has a large amount of functional component GABA, so it can be widely used as health foods and pharmaceuticals. . Furthermore, since both vitamin C and GABA components are combined in an appropriate formulation, various excellent actions (for example, a blood pressure lowering action) can be exhibited.

  The GABA-containing fermented acerola product of the present invention can be further prepared in various food forms or preparation forms according to a conventional method using an appropriate carrier or the like as necessary.

  The food form may be a beverage, a solid food, a semi-solid food or the like. Examples of the beverage include fruit juice beverages, soft drinks, and alcoholic beverages. Examples of the solid or semi-solid food include tablets, sugar-coated tablets, granules, powdered foods such as powdered drinks and powdered soups, block confectionery such as biscuits, capsules, and jelly. Moreover, various additives conventionally used for food preparation can be blended as necessary. Examples of such additives include stabilizers, pH adjusters, sugars, sweeteners, fragrances, various vitamins, minerals, antioxidants, excipients, solubilizers, binders, lubricants. , Suspending agents, wetting agents, film-forming substances, flavoring agents, flavoring agents, coloring agents, preservatives and the like.

  The food product according to the present invention preferably contains GABA in an amount of 50 mg or more per 100 g of solid content (denoted as “50 mg / 100 g solid content or more”), more preferably 75 mg / 100 g solid content or more, and 500 mg / 100 g solids. It contains vitamin C in an amount of at least minutes, more preferably at least 750 mg / 100 g solids. The food according to the present invention may be in the form of a liquid or semi-liquid such as a beverage as described above. In that case, for example, it contains GABA in an amount of 10 mg / 100 ml or more, more preferably 15 mg / 100 ml or more, and 50 mg. Vitamin C in an amount of 100 mg / 100 ml or more, more preferably 100 mg / 100 ml or more. In the present invention, analysis of ingredients in food is carried out according to the method described in “Five Amendments to Japan Food Standard Ingredients Table” (edited by the Science and Technology Agency Resource Research Committee). For example, for the solid content (g / 100 g), first, the water content (g / 100 g) is measured by an atmospheric pressure heating drying method (105 ° C., constant weight), and then the solid content (g / 100 g) = 100 g / 100 g. -Calculate the solid content by the formula of moisture content (g / 100g). The GABA content (mg / 100 ml) is measured by the method described in Test Example 1, for example. The vitamin C content (mg / 100 ml) is measured using high performance liquid chromatograph (HPLC) after derivatization by oxidizing reduced ascorbic acid to make all ascorbic acid oxidized. Based on the solid content (g / 100 g), GABA content (mg / 100 ml), and vitamin C content (mg / 100 ml) thus measured, the GABA content and vitamin C content per solid content can be determined.

  Preparation into a pharmaceutical form can be carried out according to a conventional method, and carriers and excipients, binders, preservatives, oxidation stabilizers, disintegrating agents, lubricants, corrigents, diluents that can be used at that time, It can be suitably selected from various commonly used ones. The form is not particularly limited and is appropriately selected as necessary. Generally, tablets, capsules, granules, fine granules, powders, pills, solutions, syrups, suspensions, emulsions, elixirs are used. Oral preparations such as injections, infusions, suppositories, inhalants, transdermal absorbents, transmucosal absorbents, nasal preparations, enteral preparations, patches, ointments, etc. The

  Hereinafter, GABA production reaction using acerola puree as a raw material and using yeast of Pichia or Candida, particularly Pichia anomala MR-1 will be specifically described, but the present invention is limited to these specific examples. It is not a thing.

[Test Example 1]
Acerola puree (Brazil, water 90.0%, carbohydrate 9.0%, pH 3.2) was diluted twice with sterilized distilled water, and 50 ml was poured into each sterilized 200 ml Erlenmeyer flask. Next, after adding MR-1 cells (water content 79.8%) having a predetermined concentration in Table 1 to each flask, nitrogen gas was charged at a flow rate of 300 ml / min for 1 minute. The exit of the Erlenmeyer flask was sealed with a rubber stopper, and the fermentation reaction was carried out for 2 days while shaking at 37 ° C. The reaction mixture was inactivated by heating at 85 ° C. for 15 minutes, then centrifuged, and the resulting supernatant was concentrated under reduced pressure to 25 ml each and subjected to analysis of free amino acid content including GABA components. For analysis of free amino acids, a fully automatic amino acid analyzer (JLC-500 / V) manufactured by JEOL Ltd. was used (hereinafter the same). Table 1 shows the analysis results of the GABA content in each concentrated solution obtained above.

  From the above results, it was found that a large amount of GABA can be produced even when only the MR-1 yeast of the present invention is added. Moreover, although the amount of produced | generated GABA also increased greatly with the increase in MR-1 yeast addition amount, if a microbial cell addition density | concentration is more than 15%, yeast odor will become remarkable and there exists a possibility that the flavor peculiar to acerola fruit juice may be impaired. Because there is, it is not preferable.

  The acerola puree and MR-1 cells used in the following test examples and examples are the same as those used in this test example 1 unless otherwise specified.

[Test Example 2]
A GABA production reaction was carried out under the same conditions as in Test Example 1 except that 1% concentration of glutamic acid sodium salt was added to each reaction system to prepare 25 ml of each concentrated solution. Table 2 shows the amount of GABA produced in each concentrated solution thus obtained.

  From the above results, it was found that when the sodium salt of glutamic acid was added to acerola puree, the amount of GABA produced could be increased further.

[Test Example 3]
A glutamic acid sodium salt is added to 50 ml of a reaction mixture containing 50% acerola puree and 10% MR-1 bacterial cells, respectively, and subjected to GABA production reaction under the same conditions as in Test Example 1, and then each concentrated solution 25 ml was prepared. The amount of GABA produced in each concentrated solution thus obtained is shown in Table 3.

  From the above results, when the addition concentration of MR-1 cells is 10%, the optimum addition concentration of glutamic acid sodium salt is in the range of about 0.4 to 0.8%. It was found that a larger amount of GABA could not be expected even when the addition concentration was increased.

[Test Example 4]
Each citrate-sodium phosphate buffer solution (pH 3.0 to 6.0) (0.1 M citric acid-0.2 M disodium hydrogen phosphate) was mixed with 50% acerola puree and 10 MR-1 cells. After adding a 1% concentration and 1% concentration of glutamic acid sodium salt to make a constant volume of 50 ml, GABA production reaction was performed under the same conditions as in Test Example 1, and then 25 ml of each concentrated solution was prepared. Table 4 shows the amount of GABA produced in each concentrate.

  From the above results, GABA can be produced in a large amount within the range of pH 3.0 to 5.0, but the optimum pH is about 3.0 to 4.0. It was found that the amount tended to decrease.

[Test Example 5]
A reaction mixture containing 50% acerola puree, 10% MR-1 bacterial body, 1% sodium glutamate, and 5% saccharides shown in Table 5 was prepared in 50 ml each, under the same conditions as in Test Example 1. After the GABA production reaction, 25 ml of each concentrated solution was prepared. Table 5 shows the amount of GABA produced in each concentrate.

  From the above results, it was found that even if various sugars were added to the raw material acerola puree, it was hardly expected that the amount of GABA produced would increase further.

[Test Example 6]
Prepare 50 ml each of the reaction mixture containing 50% concentration of acerola puree, 1% concentration of glutamic acid sodium salt, and 10% concentration of various yeast cells shown in Table 6 (including commercial baker's yeast manufactured by Oriental Yeast Co., Ltd.). After performing the GABA production reaction under the same conditions as in Test Example 1, 25 ml of each concentrated solution was prepared. Table 6 shows the amount of GABA produced in each concentrate.

  From the above results, yeast belonging to the genus Pichia and Candida has a high concentration of GABA producing ability, whereas yeast of the genus Saccharomyces has a low ability to produce GABA, and only about 40-50 mg / 100 ml can be seen, Moreover, even if it was the yeast of the same Pichia genus, it turned out that MR-1 yeast isolate | separated by the present inventors has especially high GABA production ability.

[Test Example 7]
50 ml of a reaction mixture containing 50% acerola puree, 1% glutamic acid sodium salt, and 10% MR-1 yeast was prepared and filled with nitrogen in the same manner as in Test Example 1 and then at 37 ° C. After performing the GABA production reaction for 24, 48 and 72 hours, 25 ml each of the concentrated solution was prepared. Table 7 shows the amount of GABA produced in each concentrate.

  From the above results, the optimal reaction time for GABA production in acerola puree by this yeast is in the range of about 48 to 72 hours. However, considering the difference between the production cost and the quality (GABA content) of the fermented acerola product, it is considered that the reaction time is preferably about 48 hours.

[Test Example 8]
Twelve 20 ml L-shaped test tubes each contain 50% acerola juice (the supernatant obtained by centrifuging the acerola puree raw material), 15% MR-1 cells, and 1% sodium glutamate 10 ml of the solution was added, and nitrogen gas was charged at a flow rate of 100 ml / min for 1 minute, and then reacted for 3 days with a temperature gradient of 15 to 60 ° C. while shaking at 20 rpm. Thereafter, separation, purification, and concentration were performed in the same manner as in Test Example 1 to prepare 5 ml of each concentrated solution. Table 8 shows the GABA concentration in each concentrate.

  From the above results, it was found that GABA was produced in a relatively large amount within a temperature range of 33 to 53 ° C, and the optimum reaction temperature was around 36 ° C. A large amount of GABA was produced even when the temperature was 40 ° C. or higher, but the fermentation reaction at “high temperature” of 40 ° C. or higher was considered in consideration of changes in the color tone of the acerola fermented liquid and the decrease in vitamin C content due to oxidation. It seems better to avoid it as much as possible.

[Example 1]
4 kg of acerola puree raw material (Brazil, water 90.0%) was poured into a 10 liter jar fermenter and diluted twice with sterilized distilled water. Since the pH of this diluted solution is 3.2, 1.2 kg (15% concentration) of MR-1 yeast cells are added and dispersed without adjusting the pH as it is, and then the same while stirring gently at 37 ° C. Fermented at temperature for 48 hours. The fermentation reaction solution was inactivated by heating at 85 ° C. for 30 minutes, and then centrifuged to obtain about 8 kg of a supernatant. These supernatants were further finely filtered and concentrated under reduced pressure to obtain 4 kg of concentrated acerola fermentation broth. The GABA content in this concentrated acerola fermentation broth was 173.2 mg / 100 ml, which was an increase of about 25 times compared to 6.8 mg / 100 ml in the acerola puree raw material before fermentation.

  In addition, Table 9 shows changes in the content of various organic components before and after fermentation. From these results, it was found that the content of monosaccharides such as glucose and fructose in the raw material was almost completely consumed by fermentation, and at the same time, organic acid components such as succinic acid and lactic acid were newly generated. . Although the vitamin C content of the functional ingredient was reduced by about 20% by fermentation for 2 days, the acerola fermented liquid was consumed due to the consumption of solids such as glucose and fructose in the acerola puree raw material. The relative content of vitamin C in the solid content of the content was rather increased compared to that in the acerola puree material.

[Example 2]
Acerola puree was fermented and purified under the same conditions as in Example 1 except that 40 g (0.5% concentration) of glutamic acid sodium salt was added to prepare 4 kg of concentrated acerola fermentation broth. The contents of various free amino acids in these acerola puree raw materials and acerola fermentation broth were measured. The results are shown in FIG. The GABA content in the acerola fermentation broth was 285.9 mg / 100 ml, which was about 42 times higher than the 6.8 mg / 100 ml in the acerola puree raw material before fermentation. In addition, the content of various other free amino acids in the obtained acerola fermented liquid also increased as shown in FIG. 1, and it was found that the increase in the contents of Ala, Pro, Asp, Ser, Leu, and Lys was particularly remarkable. . In addition, although the content of Glu is not shown in FIG. 1, about 0.36% remained based on the weight of the concentrated acerola fermentation broth.

[Example 3]
Production of GABA-containing acerola beverage (100 ml) In the following formulation, ion-exchanged water, fructose-glucose liquid sugar, purified acerola fermented liquid obtained in Example 1 or Example 2, acidulant (citric acid and sodium citrate) The mixture was sequentially put into a preparation tank, and the whole was made uniform by stirring. The product thus obtained was aseptically filtered, and hot-filled into a 100 ml container for beverages to obtain a product 1 and a product 2 respectively.

  For comparison, in place of the acerola fermented liquid, except that the acerola puree, which is the raw material for producing the acerola fermented liquid, is not fermented, and purified by the same method as the acerola fermented liquid purification method described above, An acerola beverage was produced with the same composition and the same production method, and hot-filled into a 100 ml container for beverage, thereby obtaining a comparative product 1.

(Combination)
Acerola fermentation broth: 10%
Fructose glucose liquid sugar: 11%
Citric acid: 0.3%
Sodium citrate: 0.1%
Ion exchange water: 78.6%
(All percentages are weight%)

  Table 10 shows the GABA content in each acerola beverage. The GABA contents of Examples 1 and 2 were about 25 times and 42 times higher than those of Comparative Example 1, respectively, and it was confirmed that both had a blood pressure lowering effect in animal tests. As for the flavor, both were mellow and good, but the product 2 felt a slight miscellaneous taste due to the effect of the remaining sodium glutamate salt. Therefore, it is considered that the product 1 is more suitable as a beverage.

[Example 4]
Manufacture of GABA-containing acerola jelly (150 g) Charge a suitable amount of ion-exchanged water into a jacket kneader-type blending tank equipped with a stirrer, heat it, and then use a high-speed stirring tank for sodium citrate, granulated sugar, The commercially available gelling agent (manufactured by Saneigen FFI Co., Ltd.), apple juice, pineapple juice, and purified acerola fermented liquid obtained in Example 1 or Example 2 were sequentially added. Next, the acidulant (citric acid, malic acid) was dissolved in an appropriate amount of ion-exchanged water, and similarly charged into the preparation tank. Then, after adjusting to a predetermined weight with ion-exchanged water, the mixture was homogenized by stirring, heated to 80 ° C., and stirred at the same temperature for 10 minutes. The product thus prepared was appropriately hot-packed in a jelly container using a cup filling machine, and designated as product 1 and product 2 respectively. Finally, it was immersed in warm water of 80 ° C. for 30 minutes, and heat sterilization including the container was performed.

  For comparison, instead of the acerola fermented liquid, it was purified in the same manner as in the purification method of the acerola fermented liquid except that the acerola puree, which is a raw material for producing the acerola fermented liquid, was not fermented in the same manner as in Example 3. Acerola jelly (150 g) was produced using the same composition and the same production method as described above to obtain Comparative Product 1.

(Combination)
Acerola fermentation broth: 10%
Granulated sugar: 17%
Citric acid: 0.3%
Sodium citrate 0.1%
Malic acid: 0.1%
Gelling agent (manufactured by Saneigen FFI): 1.2%
Ion exchange water: 71.3%
(All percentages are weight%)

  Table 11 shows the GABA content in each of the above acerola jelly. The GABA contents of Examples 1 and 2 are about 28 times and 48 times or more of Comparative Example 1, respectively. As for the flavor, the product 1 was mellow and good, and the product 2 had a slight taste.

[Example 5]
Manufacture of GABA-containing acerola tablets Powdered product of purified acerola fermented liquid obtained in Example 1 or Example 2, reduced maltose starch syrup, cellulose, sucrose ester, silicon dioxide, shellfish calcium with the following composition It put into the machine sequentially and mixed so that it might become uniform completely. Thereafter, tableting was performed with a tableting machine in an amount of 500 mg per tablet, and a product 1 and a product 2 were obtained. For comparison, instead of the acerola fermented liquid, it was purified in the same manner as in the purification method of the acerola fermented liquid except that the acerola puree, which is a raw material for producing the acerola fermented liquid, was not fermented in the same manner as in Example 3. 500 mg of acerola tablet per tablet was produced from the powder obtained by using the same composition and the same production method as described above to obtain Comparative Product 1.

  Table 12 shows the GABA content of each of the above acerola tablets. The GABA contents of Examples 1 and 2 are about 25 times and 43 times or more that of Comparative Example 1, respectively.

(Combination)
Acerola fermented powder: 70%
Reduced maltose starch syrup: 14%
Cellulose: 10%
Sucrose ester: 5%
Silicon dioxide: 1%
(All percentages are weight%)

It is a figure which shows the change of various free amino acid contents in the acerola processed material before and behind fermentation.

Claims (8)

The acerola fruit or processed product thereof is allowed to act on the yeast belonging to the genus Pichia or Candida or the processed product thereof having the ability to produce γ-aminobutyric acid by fermentation reaction in the presence of the acerola fruit or processed product thereof. A method for producing a fermented acerola product containing γ-aminobutyric acid, characterized by producing aminobutyric acid. The method according to claim 1 , wherein the yeast is Pichia anomala MR-1 (Accession No. FERM BP-10134) or a mutant strain thereof capable of producing γ-aminobutyric acid. The method according to claim 1 or 2 , wherein the fermentation reaction is performed under conditions of an initial pH of 3.0 to 5.0 and a temperature of 32 to 45 ° C. The method according to any one of claims 1 to 3 , wherein the fermentation reaction is performed in the presence of glutamic acid or a salt thereof. The method according to any one of claims 1 to 4 , wherein the fermentation reaction is performed in a nitrogen atmosphere. Γ- aminobutyric acid containing fermentation acerola product produced by the method according to any one of claims 1 to 5. The fermented acerola product containing γ-aminobutyric acid according to claim 6 , which is in the form of a beverage, powder, tablet or jelly. In the presence of an acerola fruit or a processed product thereof, the acerola fruit or processed product thereof is in the form of a food containing 50 mg or more of γ-aminobutyric acid per 100 g of solid content and 500 mg or more of vitamin C per 100 g of solid content . It is produced by a method for producing a fermented acerola product containing γ-aminobutyric acid, which is characterized by producing γ-aminobutyric acid by acting a yeast having the ability to produce γ-aminobutyric acid by fermentation reaction or a processed product thereof. was γ- amino butyric acid-containing fermentation acerola products.

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