Method for producing γ-aminobutyric acid (GABA)-containing powder, and product using same

Abstract

The present invention provides a method for producing a GABA-containing powder using a lactic acid bacterium, which enables the production of a GABA product having improved taste and / or odor. Specifically, the present invention provides a method for producing a γ-aminobutyric acid (GABA)-containing powder, the method comprising: (1) a fermentation step in which a lactic acid bacterium having GABA production capability is inoculated into a raw material and cultured to obtain a GAVA-containing fermented liquid; (2) a pH adjustment step in which the pH of the fermented liquid is adjusted to obtain a pH-adjusted fermented liquid; (3) a step in which the pH-adjusted fermented liquid is heated to obtain a heat-sterilized fermented liquid in which the lactic acid bacterium is sterilized; and (4) a step in which the heat-sterilized fermented liquid is treated with activated carbon.

Description

Method for producing gamma-aminobutyric acid (GABA)-containing powder and products using the same

[0001] The present invention relates to a novel method for producing γ-aminobutyric acid (GABA)-containing powder using lactic acid bacteria capable of producing γ-aminobutyric acid, which decarboxylates glutamic acid through the action of glutamic acid decarboxylase to produce GABA, and to a product containing the GABA-containing powder produced by the method.

[0002] GABA is a non-protein amino acid that is widely distributed in nature and is commonly found in vegetables, fruits, grains, etc. In vivo, it acts as an inhibitory neurotransmitter and is known to be present in particularly large amounts in the central nervous system, such as the brain and spinal cord.

[0003] It is known that GABA can be produced by fermentation using lactic acid bacteria (Patent Document 1), and fermentation products containing GABA produced by fermentation are used in many products.

[0004] Patent No. 3880820

[0005] The present invention provides a novel method for producing GABA using lactic acid bacteria, and a novel GABA-containing powder produced by such a method. According to a main aspect of the present invention, the following inventions are provided: (Item 1) A method for producing a γ-aminobutyric acid (GABA)-containing powder, comprising: (1) a fermentation step of inoculating a raw material with lactic acid bacteria capable of producing GABA and culturing the bacteria to obtain a fermentation broth containing GABA; (2) a pH adjustment step of adjusting the pH of the fermentation broth to obtain a pH-adjusted fermentation broth; (3) a step of heating the pH-adjusted fermentation broth to sterilize the lactic acid bacteria to obtain a heat-sterilized fermentation broth; and (4) a step of treating the heat-sterilized fermentation broth with activated carbon. (Item 2) A production method according to the above items, wherein the fermentation step comprises fermentation culture for 2 to 4 days. (Item 3) A production method according to any one of the above items, wherein the pH adjustment step comprises adjusting the pH to about 4 to about 7.5. (Item 4) The method according to any one of the above items, wherein the pH adjustment step comprises adjusting the pH to about 4.5 to about 7. (Item 5) The method according to any one of the above items, wherein the pH adjustment step comprises adjusting the pH to about 5 to about 6.5. (Item 6) The method according to any one of the above items, wherein the activated carbon comprises sawdust activated carbon. (Item 7) The method according to any one of the above items, wherein the lactic acid bacterium is Lactobacillus hilgardii K-3 strain (FERM BP-10487). (Item 8) The method according to any one of the above items, wherein the GABA powder has an improved taste compared to a GABA powder obtained by a similar production method except for using Lactobacillus brevis. (Item 9) The method according to any one of the above items, wherein the GABA powder has an improved odor compared to a GABA powder obtained by a similar production method except for using Lactobacillus brevis. (Item 10) The manufacturing method according to any one of the above items, further comprising drying by freeze-drying, spray-drying, vacuum drying, or any combination thereof. (Item 11) The manufacturing method according to any one of the above items, further comprising crystallization. (Item 12) A method for manufacturing a food product, comprising incorporating GABA obtained by the manufacturing method according to any one of the above items into food to obtain the food product.(Item 13) A method for producing a cosmetic, comprising blending GABA obtained by the production method described in any one of the above items into a cosmetic material to obtain a cosmetic. (Item 14) GABA powder produced by the production method described in any one of the above items. (Item 15) Food produced by the production method described in any one of the above items. (Item 16) Cosmetic produced by the production method described in any one of the above items.

[0006] In the present disclosure, it is intended that one or more of the above-described features may be provided in combination in addition to the combinations explicitly stated. Further embodiments and advantages of the present disclosure will be recognized by those skilled in the art upon reading and understanding the following detailed description as necessary. Features and significant actions and effects of the present disclosure other than those described above will become clear to those skilled in the art upon reference to the following description of the preferred embodiments of the present invention and the drawings.

[0007] The present invention can provide a novel method for producing GABA using lactic acid bacteria, which results in a GABA product with improved taste and / or odor, and a novel GABA powder produced by such a method.

[0008] FIG. 1 is a graph showing the results of bitterness, richness, saltiness, and lingering aftertaste of a GABA-containing powder produced using sawdust as activated carbon in one embodiment of the present invention. Four groups of fermented liquids were used, in which the pH of the fermented liquid after 96 hours of culture was adjusted to pH 3, pH 5.3, pH 7, or pH 9. FIG. 2 is a graph showing the results of fermentation odor, sour odor, texture, and sourness of a GABA-containing powder produced using sawdust as activated carbon in one embodiment of the present invention. Four groups of fermented liquids were used, in which the pH of the fermented liquid after 96 hours of culture was adjusted to pH 3, pH 5.3, pH 7, or pH 9. FIG. 3 is a graph showing the results of bitterness, richness, saltiness, and lingering aftertaste of a GABA-containing powder produced using coconut shell as activated carbon in one embodiment of the present invention. Four groups of fermented broths were used, in which the pH of the fermented broth after 96 hours of culture was adjusted to pH 3, pH 5.3, pH 7, or pH 9. FIG. 4 is a graph showing the results of fermentation odor, sour odor, texture, and sour taste of a GABA-containing powder produced using coconut shell as activated carbon in one embodiment of the present invention. Four groups of fermented broths were used, in which the pH of the fermented broth after 96 hours of culture was adjusted to pH 3, pH 5.3, pH 7, or pH 9. FIG. 5 is a graph showing the results of bitterness, richness, saltiness, and lingering aftertaste of a GABA-containing powder produced using coal as activated carbon in one embodiment of the present invention. Four groups of fermented broths were used, in which the pH of the fermented broth after 96 hours of culture was adjusted to pH 3, pH 5.3, pH 7, or pH 9. Fig. 6 is a graph showing the results of fermentation odor, sour odor, texture, and sour taste of GABA-containing powder produced using coal as activated carbon in one embodiment of the present invention. Four groups of fermentation liquids were used, in which the pH of the fermentation liquid after 96 hours of culture was adjusted to pH 3, pH 5.3, pH 7, or pH 9. Fig. 7 is a graph showing the results of bitterness, richness, saltiness, and lingering aftertaste of GABA-containing powder produced using sawdust, coconut shell, and coal as activated carbon in one embodiment of the present invention. Fermentation liquids with a pH of 6 to 6.3 after 96 hours of culture were used.8 is a graph showing the results of fermentation odor, sour odor, texture, and sour taste of GABA-containing powder produced using sawdust, coconut shell, and coal as activated carbon in one embodiment of the present invention. The fermented liquid used had a pH of 6 to 6.3 after 96 hours of cultivation.

[0009] The present disclosure will now be described with reference to the best mode. Throughout this specification, singular expressions should be understood to include the plural concept unless otherwise specified. Therefore, singular articles (e.g., "a," "an," "the," etc. in English) should be understood to include the plural concept unless otherwise specified. Furthermore, it should be understood that terms used in this specification are used in the sense commonly used in the art unless otherwise specified. Therefore, unless otherwise defined, all technical and scientific terms used in this specification have the same meaning as commonly understood by those skilled in the art to which this disclosure belongs. In the event of conflict, the present specification (including definitions) will prevail.

[0010] (Definitions) The definitions of terms particularly used in this specification and / or basic technical content will be explained as appropriate below.

[0011] As used herein, "about" means ±10% of the preceding numerical value.

[0012] As used herein, "room temperature" refers to a temperature between about 20°C and about 40°C.

[0013] In this specification, "standing still" refers to being left in a quiet environment in a stationary state, without being subjected to external artificial physical stimuli such as vibrations or sound waves.

[0014] As used herein, "γ-aminobutyric acid (GABA)-containing powder" refers to any powder in which the main component is GABA. When a powder is "mainly composed of a certain component," this means that the powder contains 20% by weight or more of that component.

[0015] As used herein, substance A has "improved taste quality" compared to substance B means that, when the results of sensory evaluations of substance A and substance B relative to a reference substance are averaged by multiple evaluators, substance A has a superior evaluation result to substance B. In the present invention, the sensory evaluation is performed by multiple evaluators using a VAS questionnaire format to evaluate the strength of the taste compared to the reference using a five-point scale of -2, -1, 0, +1, and +2, and the results of all evaluators are averaged. The taste quality in the present invention may be, but is not limited to, saltiness, a lingering aftertaste, or bitterness.

[0016] In this specification, substance A having an "improved odor" compared to substance B means that, when the results of sensory evaluation of the odors of substance A and substance B relative to a reference substance are averaged, substance A has a superior evaluation result to substance B. The odor in the present invention may be, but is not limited to, a yeasty odor, a fermented odor, or a sour odor.

[0017] As used herein, "adjusting" pH generally refers to changing the pH of a composition (e.g., a liquid) by adding an acid or a base. This includes adjusting from an acidic to a neutral or alkaline state, and adjusting from an alkaline state to a neutral or acidic state. The pH can be adjusted using any acid or alkali, such as hydrochloric acid, sodium hydroxide, or sodium bicarbonate.

[0018] As used herein, "activated carbon treatment" refers to a process in which a solution is brought into contact with activated carbon to remove impurities such as particles contained in the solution by adsorbing them onto the activated carbon. Specifically, this is a process in which the solution to be treated is mixed with or passed through activated carbon. In this activated carbon treatment, the amount and concentration of the solution to be treated, the type, amount, contact time, temperature, etc. of activated carbon used can be appropriately set. Examples of activated carbon include sawdust, coconut shells, and coal. After treatment with activated carbon, the activated carbon can be removed by known solid-liquid separation means such as filtration or centrifugation to obtain the desired activated carbon-treated solution.

[0019] In this specification, "or" is used when "at least one or more" of the items listed in the sentence can be employed. The same applies to "alternative." In this specification, when it is specified that "within a range" of "two values," the range includes the two values ​​themselves.

[0020] (Lactic Acid Bacteria and Fermentation Thereof) The lactic acid bacterium of the present invention may be any lactic acid bacterium whose growth is promoted by the addition of glutamic acid or a salt thereof to a culture medium and which has the ability to produce γ-aminobutyric acid by the action of glutamic acid decarboxylase. In a typical embodiment, the lactic acid bacterium of the present invention is capable of producing 45 g / L or more of γ-aminobutyric acid when cultured at 30°C for 48 hours in a liquid medium containing 100 g of sodium glutamate per liter. In a particularly preferred embodiment, the lactic acid bacterium of the present invention is the K-3 strain (FERM BP-10487) belonging to Lactobacillus hilgardii.

[0021] The mycological properties of the K-3 strain are shown in Table 1.

[0022] In the present invention, lactic acid bacteria are used to produce a fermented food containing γ-aminobutyric acid by inoculating a fermentation raw material containing glutamic acid or a salt thereof with the lactic acid bacteria and culturing the lactic acid bacteria. As the lactic acid bacteria, any of freeze-dried cells, frozen-preserved strains, and liquid culture solutions may be used, but it is preferable to use lactic acid bacteria that have been pre-cultured in a fermentation solution or liquid medium containing 1% or more glutamic acid from the day before use.

[0023] The glutamic acid used in the production method of the present invention refers chemically to L-glutamic acid, a type of amino acid. It may be glutamic acid, monosodium glutamate, or other glutamates, which are food additives used as seasonings, or glutamic acid obtained by hydrolyzing food proteins with acids or enzymes. Foods containing free glutamic acid, such as seasonings, processed seafood products, and tomatoes, may also be used as is. However, if a food product containing a higher amount of γ-aminobutyric acid is desired, it is necessary to use a fermentation raw material containing a higher amount of glutamic acid.

[0024] In order to grow lactic acid bacteria, the fermentation medium preferably contains, in addition to the glutamic acid described above, carbohydrates such as glucose, fructose, and maltose, and foods containing vitamins and minerals such as yeast extract and meat extract.Furthermore, food additives such as emulsifiers, stabilizers, and pH adjusters can be used.

[0025] The size and material of the container used for the fermentation process can be any container that can be washed and heat sterilized and can be used for food production, but it is preferable that the container has a structure that makes it difficult for bacteria to enter.

[0026] The fermentation temperature is preferably about 20°C to about 30°C, and the fermentation time is, for example, about 48 to about 96 hours. When fermentation is performed using a fermentation medium containing a high concentration of monosodium glutamate of 5% by weight or more, an initial pH of 4.5 to 5.5, at which bacterial growth is most promoted, is most preferred. In one embodiment, the fermentation time is preferably about 96 hours.

[0027] In the fermentation of a raw material containing glutamic acid or its salts using lactic acid bacteria, for example, when the K-3 strain was cultured in a liquid medium containing 4 wt% sodium glutamate, the amount of γ-aminobutyric acid produced after 72 hours was approximately 2 wt% at both the initial pH of 6.3 and the initial pH of 5.0. In contrast, when the K-3 strain was cultured in a liquid medium containing 10 wt% sodium glutamate, the amount of γ-aminobutyric acid produced after 72 hours was approximately 3 wt% at the initial pH of 6.3, but approximately 5 wt% at the initial pH of 5.0. Furthermore, the pH of the fermentation broth increases with the growth of the K-3 strain, reaching a neutral pH of 7 to 8 24 to 48 hours after the start of fermentation. Therefore, the pH of the fermentation broth is adjusted back to approximately 5 approximately 24 hours after the start of fermentation, and the fermentation process is continued. This further promotes the growth of lactic acid bacteria and increases the rate and efficiency of γ-aminobutyric acid production.

[0028] (pH Adjustment of Fermentation Broth) The present invention involves adjusting the pH of a γ-aminobutyric acid-containing fermentation broth obtained by culturing lactic acid bacteria after a predetermined period of fermentation to obtain a pH-adjusted fermentation broth. The pH adjustment can be performed using any acid or alkali, including, but not limited to, hydrochloric acid, sodium hydroxide, and sodium bicarbonate. All of these pH adjusters are approved for use in food manufacturing processes and are known not to affect the safety of the food to which they are added. Without intending to be bound by theory, the improvement in taste and odor achieved by the production method of the present invention compared to Lactobacillus brevis (NBRC 12005) is believed to be due to the GABA-producing activity of the lactic acid bacteria (Lactobacillus hilgardii K-3 strain (FERM BP-10487)). Therefore, it is understood that the improvement in taste and odor achieved by the present invention does not depend on the type of pH adjuster, but rather on the type of lactic acid bacteria and the processing conditions of the fermentation broth. In a preferred embodiment, the γ-aminobutyric acid-containing fermentation liquor obtained by culturing lactic acid bacteria after the heat sterilization step can be adjusted to a pH of about 4 to about 7.5. In a more preferred embodiment, the fermentation liquor of the present invention can be adjusted to a pH of about 4.5 to about 7. In an even more preferred embodiment, the fermentation liquor of the present invention can be adjusted to a pH of about 5 to about 6.5. In a most preferred embodiment, the fermentation liquor of the present invention can be adjusted to a pH of about 5.3.

[0029] (Heating for killing lactic acid bacteria and stopping fermentation (heat sterilization step)) In the present invention, fermentation is carried out for a predetermined period of time, the pH is adjusted, and then heating (heat sterilization step) is carried out to kill the lactic acid bacteria and stop fermentation. The heat sterilization step can be carried out under any conditions that kill the lactic acid bacteria in the fermentation liquid. Since lactic acid bacteria are killed by heating at about 75°C or higher for about 15 minutes, the conditions for the heat sterilization step of the present invention can be heating at a temperature of about 75°C or higher for about 15 minutes or more.

[0030] In one embodiment, the heat sterilization step of the present invention can be carried out by heating at about 75°C to about 120°C for about 30 minutes.

[0031] (Mixing with an excipient after fermentation) In one embodiment, after the heat sterilization step, the fermented broth containing γ-aminobutyric acid obtained by culturing lactic acid bacteria is mixed with an excipient to obtain a mixed broth. Excipients added to the fermented broth at this stage include, but are not limited to, dextrin, starch, cyclodextrin, maltodextrin, gum arabic, etc. All of these excipients are known to be tasteless and odorless and do not affect the taste or odor of the food to which they are added. In a preferred embodiment, the excipient added to the fermented broth at this stage is starch or dextrin.

[0032] In one embodiment, when mixing the excipient with the fermentation broth, the mixture may be heated (heating and mixing step) to promote mixing of the excipient with the fermentation broth. The heating and mixing step may be performed at a temperature that can at least partially dissolve the excipient and promote mixing of the excipient with the fermentation broth. Excipients such as starch, dextrin, and cyclodextrin can generally be promoted by heating to 40°C or higher.

[0033] In one embodiment, the heating and mixing step of the present invention is carried out by heating at about 60°C to about 120°C for 30 minutes, and in a preferred embodiment, it can be carried out by heating at about 75°C to about 110°C for 30 minutes or more.

[0034] The fermentation liquor of the present invention may be subjected to processing steps such as filtration, concentration, etc. in addition to mixing with an excipient. Typically, after the heat sterilization step, the fermentation liquor may be filtered and / or concentrated to further increase the γ-aminobutyric acid content before use.

[0035] (Activated carbon treatment of heat-sterilized fermentation liquor) In the present invention, post-fermentation treatments include, but are not limited to, concentration of the heat-sterilized fermentation liquor, removal of impurities, clarification, desalting, decolorization, enzyme treatment, etc. In a typical embodiment, the present invention comprises treating the heat-sterilized fermentation liquor with activated carbon. In one embodiment, membrane treatment, column purification, centrifugation, or the like can be performed preliminarily before or after activated carbon treatment for desalting, decolorization, impurity removal, etc.

[0036] The activated carbon treatment can be carried out by mixing or passing the heat-sterilized fermentation liquor through activated carbon. Examples of activated carbon include, but are not limited to, activated carbon derived from sawdust, coconut shells, and coal. In one embodiment, the amount, pH, and concentration of the fermentation liquor to be treated, the type and amount of activated carbon, contact time, temperature, and other factors can be appropriately set. In one embodiment, the type of activated carbon can be changed depending on the type of taste and / or odor desired to be improved in the final γ-aminobutyric acid powder. The type of activated carbon can also be changed depending on the properties (amount, pH, concentration, etc.) of the fermentation liquor to be treated with activated carbon. In a preferred embodiment, the activated carbon used in the activated carbon treatment is derived from sawdust.

[0037] In one embodiment, the sawdust, coconut shells, or coal used as activated carbon may be of any type, and may be in powder, granular, or pellet form. In the present invention, by treating the heat-sterilized fermentation liquid with activated carbon, the taste and / or odor of the final GABA-containing powder can be improved, including bitterness, richness, saltiness, lingering aftertaste, fermentation odor, sour odor, texture, and acidity. For example, key parameters used to evaluate the performance and quality of activated carbon include adsorption capacity, internal surface area, pore size distribution, particle size, particle size, surface functional groups, specific surface area, and iodine value. Regardless of the activated carbon used, improved taste and / or odor can be achieved. Without being bound by theory, this is thought to be due to the fact that impurities from the GABA production process are adsorbed into the fine pores of the activated carbon by treating the fermentation liquid with activated carbon.

[0038] In one embodiment, in the method of the present invention, the specific surface area of ​​the activated carbon is about 800 to about 1100 m ２ / g, preferably about 900 to about 1000 m ２In the method of the present invention, activated carbon having an iodine value of about 900 to about 1300 mg / g, preferably about 1000 to about 1200 mg / g, can be used. In the method of the present invention, activated carbon having a particle size of about 75 μm or less, preferably about 45 μm or less, and more preferably at least about 90% of the particles having a particle size of about 45 μm or less can be used.

[0039] As described above, in one embodiment of the present invention, a pH-adjusted fermented liquor can be obtained by adjusting the pH of a γ-aminobutyric acid-containing fermented liquor obtained by culturing lactic acid bacteria. In a preferred embodiment, the pH of the fermented liquor is adjusted to a weak acidity, for example, a pH of about 4.5 to about 7, a pH of about 5 to about 6.5, or a pH of about 5.3, and then the pH-adjusted fermented liquor is heat-sterilized and further treated with sawdust-derived activated carbon, thereby achieving improvements in the taste and / or odor of the final GABA-containing powder, including bitterness, richness, saltiness, lingering aftertaste, fermentation odor, sour odor, texture, and sourness.

[0040] After treatment with activated carbon, the activated carbon can be removed by known solid-liquid separation means such as filtration or centrifugation to obtain the desired activated carbon-treated solution.

[0041] Filtration can be carried out using ordinary food processing filtration equipment using filter paper, filter cloth, etc., and filter aids such as diatomaceous earth, cellulose, activated carbon, etc. The concentration step can be carried out using equipment such as a vacuum concentrator, a reduced pressure concentrator, a distillation still, or a freeze concentrator, or the water in the fermentation liquid can be evaporated in a pot over a fire.

[0042] These processing steps may occur before or after mixing with the excipients. Typically, the processing steps occur after mixing with the excipients.

[0043] (Standing After Heating and Mixing Step) In one embodiment, after the heating and mixing step, the fermentation liquid or treated liquid can be left standing under specific conditions.

[0044] In a preferred embodiment, in the production method of the present invention, after the heating and mixing step, the mixed liquid may be allowed to stand for about 10 minutes or more, about 20 minutes or more, more preferably about 30 minutes or more, before being subjected to the subsequent drying step. In a preferred embodiment, in the production method of the present invention, after the heating and mixing step, the mixed liquid may be allowed to stand for about 100 hours or less, about 48 hours or less, about 24 hours or less, more preferably about 18 hours or less, before being subjected to the subsequent drying step. In a more preferred embodiment, in the production method of the present invention, after the heating and mixing step, the mixed liquid may be allowed to stand for about 20 minutes to about 48 hours, about 20 minutes to about 24 hours, about 20 minutes to about 18 hours, about 30 minutes to about 48 hours, about 30 minutes to about 24 hours, or about 30 minutes to about 18 hours, before being subjected to the subsequent drying step.

[0045] The standing step may be carried out at any temperature, but is preferably carried out at room temperature.

[0046] (Drying and crystallization) The fermentation liquid after the activated carbon treatment can be heated, filtered, concentrated, desalted as needed, and then subjected to a drying process to obtain a GABA-containing powder containing GABA.The drying process can be carried out by an efficient and hygienic method known in the art, such as spray drying, freeze drying, or vacuum drying, or these drying methods can be used in combination.In a typical embodiment of the present invention, the drying process can be carried out by spray drying.

[0047] A sterilization step may be further carried out before or after a drying step such as spray drying or freeze drying.

[0048] In one embodiment of the present invention, the fermentation liquor that has been treated with activated carbon can be cooled to crystallize, and the crystals can be collected and then subjected to a drying process. Crystallization of the fermentation liquor can be carried out by techniques known in the art, such as cooling crystallization (reducing solubility by cooling), evaporation crystallization (evaporating a solution to increase the solute concentration), antisolvent addition crystallization (reducing solubility by adding a solvent in which the solute is poorly soluble), reactive crystallization (inducing a chemical reaction using a precipitant, reactive gas, etc.), and any combination thereof.

[0049] (Preferred embodiment) The GABA-containing powder obtained by the manufacturing method of the present invention may have a GABA content of about 20% by weight, about 30% by weight, about 40% by weight, about 50% by weight, about 60% by weight, about 70% by weight, about 80% by weight, about 90% by weight, or more, based on the solid content of GABA. The GABA content in the GABA-containing powder may vary depending on factors such as the fermentation medium used, culture conditions, type and strain of microorganism, concentration and purification methods, etc.

[0050] The GABA-containing powder of the present invention may contain, in addition to GABA, other components such as other amino acids and sugars, fermentation medium components, and microorganisms or parts thereof.

[0051] In a particularly preferred embodiment, in the production method of the present invention, the fermentation liquid is adjusted to a pH of about 4 to about 7.5, preferably about 4.5 to about 7, more preferably about 5 to about 6.5 in the pH adjustment step, and the heat-sterilized fermentation liquid can be treated with activated carbon using sawdust. By producing a GABA-containing powder in this manner, the resulting GABA-containing powder can achieve improved taste and / or odor, including bitterness, richness, saltiness, lingering aftertaste, fermented odor, sour odor, texture, and sourness, preferably improved taste and odor.

[0052] In a preferred embodiment, a GABA-containing powder obtained using the Lactobacillus hilgardii K-3 strain according to the present invention has a more favorable evaluation in a sensory evaluation of taste and / or odor relative to a standard, compared to a GABA-containing powder (comparative GABA-containing powder) obtained under similar conditions except for using Lactobacillus brevis (e.g., NBRC 12005). In a specific embodiment, the GABA-containing powder of the present invention may have a sensory evaluation of bitterness that is 0.4 or more, preferably 0.5 or more, and more preferably 0.6 or more, better than the comparative GABA-containing powder. In a specific embodiment, the GABA-containing powder of the present invention may have a sensory evaluation of kokumi that is 0.4 or more, preferably 0.5 or more, and more preferably 0.6 or more, better than the comparative GABA-containing powder. In a specific embodiment, the GABA-containing powder of the present invention may have a sensory evaluation of saltiness that is 0.3 or more, preferably 0.4 or more, and more preferably 0.5 or more, better than the comparative GABA-containing powder. In certain embodiments, the GABA-containing powder of the present invention may have a sensory evaluation of lingering aftertaste that is 0.3 or more, preferably 0.4 or more, more preferably 0.5 or more better than the comparative GABA-containing powder. In certain embodiments, the GABA-containing powder of the present invention may have a sensory evaluation of fermented odor that is 0.4 or more, preferably 0.5 or more, more preferably 0.6 or more better than the comparative GABA-containing powder. In certain embodiments, the GABA-containing powder of the present invention may have a sensory evaluation of sour odor that is 0.1 or more, more preferably 0.2 or more better than the comparative GABA-containing powder. In certain embodiments, the GABA-containing powder of the present invention may have a sensory evaluation of texture that is 0.4 or more, preferably 0.5 or more, more preferably 0.6 or more better than the comparative GABA-containing powder. In certain embodiments, the GABA-containing powder of the present invention may have a sensory evaluation of sour taste that is 0.4 or more, preferably 0.5 or more, more preferably 0.6 or more better than the comparative GABA-containing powder.

[0053] Preferably, the GABA-containing powder of the present invention may have better sensory evaluations than the comparative GABA-containing powder in any one, two, three, four, five, six, seven, or eight of bitterness, richness, saltiness, lingering aftertaste, fermented smell, sour smell, texture, and sourness.

[0054] (Uses) The GABA-containing powder obtained by the manufacturing method of the present invention can be incorporated into food products. When the GABA-containing powder obtained by the manufacturing method of the present invention is used in food products, the improved taste and odor can be advantageous.

[0055] Foods containing GABA-containing powder obtained by the manufacturing method of the present invention can further contain optional ingredients commonly used in food manufacturing. Examples of such ingredients include proteins, carbohydrates, fats, nutrients, seasonings, and flavorings. Examples of carbohydrates include common sugars such as monosaccharides (e.g., glucose, fructose, etc.); disaccharides (e.g., maltose, sucrose, oligosaccharides, etc.); and polysaccharides (e.g., dextrin, cyclodextrin, etc.), as well as sugar alcohols such as xylitol, sorbitol, and erythritol. Examples of flavorings include natural flavors (e.g., thaumatin, stevia extract, etc.) and synthetic flavors (e.g., saccharin, aspartame, etc.). Other additives commonly added to foods, such as excipients, binders, disintegrants, lubricants, stabilizers, flavorings, odorants, pH adjusters, and colorants, may also be used.

[0056] The food in the present invention may be a solid food or a liquid food or beverage. Examples of solid foods include, but are not limited to, dried foods, supplements, etc., such as jelly, yogurt, frozen desserts, candy, tablets, chocolate, gum, crackers, biscuits, cookies, cakes, bread, etc. Furthermore, examples of liquid foods and beverages include, but are not limited to, soft drinks, sports drinks, mineral water, carbonated drinks, tea (green tea, oolong tea, black tea, herbal tea, etc.), mineral water, concentrated fruit juice, reconstituted juice from concentrate, straight juice, mixed fruit juice, fruit juice with pulp, fruit juice drink, fruit and vegetable mixed juice, vegetable juice, milk, dairy drink, cocoa drink, powdered drink, etc.

[0057] The GABA-containing powder obtained by the manufacturing method of the present invention can also be incorporated into cosmetics. When the GABA-containing powder obtained by the manufacturing method of the present invention is used in cosmetics, the improved odor can be advantageous.

[0058] Cosmetics containing GABA-containing powder obtained by the manufacturing method of the present invention can be blended with any of the ingredients commonly used in cosmetic manufacturing.Such ingredients include, for example, oils and waxes such as macadamia nut oil, avocado oil, corn oil, olive oil, rapeseed oil, sesame oil, castor oil, safflower oil, cottonseed oil, jojoba oil, coconut oil, palm oil, liquid lanolin, hydrogenated coconut oil, hydrogenated oil, Japan wax, hydrogenated castor oil, beeswax, candelilla wax, carnauba wax, ivotaro wax, lanolin, reduced lanolin, hard lanolin, jojoba wax, etc.; liquid paraffin, squalane, pristane, ozokerite, paraffin, ceteth-1 ... hydrocarbons such as resin, petrolatum, and microcrystalline wax; higher fatty acids such as oleic acid, isostearic acid, lauric acid, myristic acid, palmitic acid, stearic acid, behenic acid, and undecylenic acid; higher alcohols such as cetyl alcohol, stearyl alcohol, isostearyl alcohol, behenyl alcohol, octyldodecanol, myristyl alcohol, and cetostearyl alcohol; compounds such as cetyl isooctanoate, isopropyl myristate, hexyldecyl isostearate, diisopropyl adipate, di-2-ethylhexyl sebacate, cetyl lactate, diisostearyl malate, ethylene glycol di-2-ethylhexanoate, neopentyl glycol dicaprate, glycerin di-2-heptylundecanoate, glycerin tri-2-ethylhexanoate, trimethylolpropane tri-2-ethylhexanoate, trimethylolpropane triisostearate, and pentane erythritol tetra-2-ethylhexanoate. Oils such as synthetic ester oils; chain polysiloxanes such as dimethylpolysiloxane, methylphenylpolysiloxane, and diphenylpolysiloxane; cyclic polysiloxanes such as octamethylcyclotetrasiloxane, decamethylcyclopentasiloxane, and dodecamethylcyclohexanesiloxane; and silicone oils such as modified polysiloxanes such as amino-modified polysiloxane, polyether-modified polysiloxane, alkyl-modified polysiloxane, and fluorine-modified polysiloxane may also be used.

[0059] The present disclosure has been described above by showing preferred embodiments for ease of understanding. The present disclosure will be described below based on examples. However, the above description and the following examples are provided for illustrative purposes only and are not intended to limit the present disclosure. Therefore, the scope of the present disclosure is not limited to the embodiments or examples specifically described herein, but is limited only by the scope of the claims.

[0060] (Example 1: Study of pH adjustment and activated carbon treatment conditions after raw material fermentation) Lactobacillus hilgardii K-3 strain (FERM BP-10487) and Lactobacillus brevis NBRC 12005 strain were used as lactic acid bacteria, and GABA-containing powders were produced under the same conditions for each strain. The specific production conditions are as follows.

[0061] First, lactic acid bacteria were cultured at 30°C for 96 hours. After 96 hours of culture, the pH of the fermented broth was adjusted to pH 3, pH 5.3, pH 7, or pH 9 to obtain four groups of fermented broth. HCl was used to adjust the acidic pH (pH 3 and pH 5.3), and NaOH was used to adjust the basic pH (pH 7 and pH 9).

[0062] After adjusting the pH, the lactic acid bacteria were heat-sterilized at 97°C for 30 minutes. The heat-sterilized fermentation liquid was treated with activated carbon and filtered. For the activated carbon treatment, each pH-adjusted group was further divided into three groups, and each group was treated with three types of activated carbon: sawdust (Taiko SW50) (manufactured by Futamura Chemical Co., Ltd.), coconut shell (Shirasagi WP) (manufactured by Osaka Gas Chemicals Co., Ltd.), or coal (Shirasagi DO-5) (manufactured by Osaka Gas Chemicals Co., Ltd.). Each group was then further heat-sterilized and desalted, concentrated, crystallized, and then dried to obtain a GABA-containing powder.

[0063] For the GABA-containing powder obtained under each condition (24 combinations in total, 2 types of lactic acid bacteria × 4 types of pH adjustment × 3 types of activated carbon), three evaluators actually ate a small amount of the GABA-containing powder and evaluated it for bitterness, richness, saltiness, lingering aftertaste, texture, and sourness.In addition, for the GABA-containing powder obtained under each condition, three evaluators smelled the bag containing the powder and evaluated it for fermentation odor and sour odor.The evaluators used the commercially available "Lactogaban" (Pharma Foods Co., Ltd.) as a standard and checked the strength of each item in a VAS questionnaire format with a 5-point rating of -2, -1, 0, +1, +2.The average value of the three evaluators was taken as the result of the sensory evaluation.

[0064] The results for bitterness, richness, saltiness, and lingering aftertaste when sawdust was used as activated carbon are shown in Figure 1, and the results for fermentation odor, sour odor, texture, and sourness are shown in Figure 2.

[0065] The results for bitterness, richness, saltiness, and lingering aftertaste when coconut shell was used as activated carbon are shown in Figure 3, and the results for fermentation odor, sour odor, texture, and sourness are shown in Figure 4.

[0066] The results for bitterness, richness, saltiness, and lingering aftertaste when coal was used as activated carbon are shown in Figure 5, and the results for fermentation odor, sour odor, texture, and sourness are shown in Figure 6.

[0067] As shown in Figures 1 to 6, when the K-3 strain was used, adjusting the pH of the raw material after fermentation to pH 5.3 to 7 was superior to adjusting the pH to pH 3 or 9 in the evaluations of bitterness, lingering aftertaste, fermentation odor, sour odor, and sourness. In particular, when sawdust was used as activated carbon, the results were superior in the evaluations of bitterness, richness, saltiness, lingering aftertaste, fermentation odor, sour odor, and sourness. As is clear from Figures 1 to 6, this phenomenon was not observed in the B. brevis strain, a common lactic acid bacterium also capable of producing GABA, and was surprising in that it was unique to the K-3 strain.

[0068] (Example 2: Examination of activated carbon treatment conditions in other production methods) Since Example 1 suggested that it is preferable to adjust the pH to weakly acidic after fermentation of the raw material, preferable activated carbon treatment conditions when the production method is changed were examined.

[0069] GABA-containing powders were produced using Lactobacillus hilgardii K-3 strain (FERM BP-10487) and Lactobacillus brevis NBRC 12005 strain as lactic acid bacteria under the same conditions. The specific production conditions are as follows:

[0070] First, the lactic acid bacteria were cultured for 96 hours at 30° C. After 96 hours of culture, the pH of the fermentation liquid was confirmed to be 6 to 6.3.

[0071] The lactic acid bacteria were then heat-sterilized at 97°C for 30 minutes. The fermentation liquid after heat sterilization was concentrated, treated with activated carbon, and filtered. For the activated carbon treatment, the fermentation liquid was divided into three groups, and each was treated with three types of activated carbon: sawdust (Shirasagi A) (manufactured by Futamura Chemical Co., Ltd.), coconut shell (Shirasagi WP) (manufactured by Osaka Gas Chemicals Co., Ltd.), or coal (Shirasagi DO-5) (manufactured by Osaka Gas Chemicals Co., Ltd.). After activated carbon treatment, the fermentation liquid of each group was concentrated and dried under reduced pressure to obtain a GABA-containing powder.

[0072] The GABA-containing powder obtained under each condition (two types of lactic acid bacteria × three types of activated carbon, a total of six cases) was evaluated by three evaluators who actually ate a small amount of the GABA-containing powder for bitterness, richness, saltiness, lingering aftertaste, texture, and sourness.In addition, the GABA-containing powder obtained under each condition was evaluated by three evaluators who smelled the bag containing the powder for fermentation odor and sour odor.The evaluators used a commercially available GABA product (manufactured by Pharma Foods Co., Ltd.) as the standard and checked the strength of each item in a VAS questionnaire format using a five-point scale of -2, -1, 0, +1, +2.The average value of the three evaluators was used as the result of the sensory evaluation.

[0073] The results for bitterness, richness, saltiness, and lingering aftertaste when the pH of the fermentation liquid after 96 hours of culture was weakly acidic are shown in Figure 7, and the results for fermentation odor, sour odor, texture, and sourness are shown in Figure 8.

[0074] As shown in Figures 7 and 8, when the pH of the raw material after fermentation using the K-3 strain was weakly acidic (pH 6 to 6.3), the activated carbon made from sawdust was found to improve bitterness, richness, saltiness, lingering aftertaste, fermentation odor, and sour odor more than the other two types of activated carbon. As is clear from Figures 7 and 8, this phenomenon was not observed with the K-3 strain, a common lactic acid bacterium that also has the ability to produce GABA, and was surprising in that it was unique to the K-3 strain.

[0075] As demonstrated by these results, improving taste and odor was achieved by adjusting the pH of the raw material after fermentation to 5.3-7 using the K-3 strain. Furthermore, it was demonstrated that these characteristics can be achieved by using various activated carbons, including sawdust, depending on the type of taste and odor, and are not dependent on the type of activated carbon. (Note) While the present disclosure has been illustrated using preferred embodiments thereof, it is understood that the scope of the present disclosure should be interpreted solely by the claims. It is understood that the patents, patent applications, and other documents cited herein are incorporated by reference in their entirety as if specifically set forth herein. This application claims priority to Japanese Patent Application No. 2024-7402, filed with the Japan Patent Office on January 22, 2024, the contents of which are incorporated by reference in their entirety as if they constitute the subject matter of this application.

[0076] A novel method for producing a GABA-containing powder and an article containing the GABA-containing powder produced by the method are provided.

[0077] Lactobacillus hilgardii K-3 strain FERM BP-10487

[0078]

Claims

1. A method for producing a γ-aminobutyric acid (GABA)-containing powder, comprising: (1) an fermentation step of inoculating and culturing lactic acid bacteria having the ability to produce GABA in a raw material to obtain a fermentation broth containing GABA; (2) a pH adjustment step of adjusting the pH of the fermentation broth to obtain a pH-adjusted fermentation broth; (3) a step of heating the pH-adjusted fermentation broth to obtain a heat-sterilized fermentation broth in which the lactic acid bacteria are sterilized; and (4) a step of treating the heat-sterilized fermentation broth with activated carbon.

2. The production method according to claim 1, wherein the fermentation step includes fermentative culturing for 2 to 4 days.

3. The production method according to claim 1, wherein the pH adjustment step includes adjusting the pH to about 4 to about 7.

5.

4. The production method according to claim 1, wherein the pH adjustment step includes adjusting the pH to about 4.5 to about 7.

5. The production method according to claim 1, wherein the pH adjustment step includes adjusting the pH to about 5 to about 6.

5.

6. The production method according to claim 1, wherein the activated carbon includes sawdust activated carbon.

7. The production method according to claim 1, wherein the lactic acid bacteria are Lactobacillus hilgardii K-3 strain (FERM BP-10487).

8. The production method according to claim 1, wherein the GABA powder has improved taste quality as compared with the GABA powder obtained by producing in the same manner except using Lactobacillus brevis.

9. The production method according to claim 1, wherein the GABA powder has improved odor as compared with the GABA powder obtained by producing in the same manner except using Lactobacillus brevis.

10. The production method according to claim 1, further including drying by freeze-drying, spray-drying, vacuum drying, or any combination thereof.

11. The production method according to claim 1, further including crystallization.

12. A method for producing a food, comprising incorporating the GABA obtained by the production method according to any one of claims 1 to 11 into a food to obtain a food product.

13. A method for producing a cosmetic, comprising incorporating the GABA obtained by the production method according to any one of claims 1 to 11 into a cosmetic to obtain a cosmetic product.

14. A GABA powder produced by the production method according to any one of claims 1 to 11.

15. A food product produced by the production method according to claim 12.

16. A cosmetic product produced by the production method according to claim 13.

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