JP6786223B2 - Lactic acid bacteria and a method for producing lactic acid fermented sake lees fermented extract using it - Google Patents

Description

The present invention relates to lactic acid bacteria and a method for producing a lactic acid fermented sake lees fermented extract using the same.

Sake lees contain many nutrients such as various amino acids, vitamins, organic acids, proteins, and various sugars, which are ingredients derived from rice, rice jiuqu, and yeast, and active ingredients such as delicious ingredients. Nevertheless, it is currently used only as a food material for pickles such as Narazuke, amazake, and Jiuqu shochu, and it cannot be said that these active ingredients are fully utilized.

Regarding the effective use of sake lees, fermentation of sake lees with lactic acid bacteria is being studied. For example, Patent Document 1 discloses a functional food containing γ-aminobutyric acid (GABA), which is obtained by fermenting sake lees only with lactic acid bacteria or by coexisting with aspergillus, to suppress an increase in blood pressure.

However, sake lees, which is a brewing by-product, still needs to be further developed for use, and it is desired to obtain a new functional material.

Patent Document 2 discloses a method for producing an L-ornithine-containing product, which ferments a protein / or peptide-containing product containing an amino acid constituting L-arginine using a lactic acid bacterium of the genus Lactococcus or the genus Lactobacillus. However, Patent Document 2 does not describe that sake lees are used as a raw material.

Patent Document 3 discloses a method for producing a D-amino acid using a microorganism belonging to Lactobacillus gasseri. However, Patent Document 3 does not describe that sake lees are used as a raw material. Black vinegar is known as a food containing a high content of D-serine, but according to Non-Patent Document 1, the maximum content is 307 μM (about 32 mg / L).

Japanese Unexamined Patent Publication No. 2006-9413 JP-A-2009-112205 Japanese Unexamined Patent Publication No. 2015-47120

Trace Nutrients Research 29: 62-66 (2012)

An object of the present invention is to provide a novel fermented sake lees extract made from sake lees, a method for producing the same, and a microorganism that can be used in the method for producing the same, in order to effectively utilize sake lees.

As a result of diligent research to solve the above problems, the present inventors have further fermented a fermented sake lees extract with a microorganism of the genus Lactobacillus to obtain an extract containing a high content of ornithine and / or D-amino acid. The present invention was completed by finding that it can be produced.

That is, the present invention
(1) Lactobacillus is a fermented liquor extract obtained by subjecting liquor to starch-degrading enzyme treatment and / or proteolytic enzyme treatment and fermentation treatment with yeast, and then performing solid-liquid separation to remove solid components. A method for producing a fermented lactic acid liquor extract, which comprises fermenting with one or more Lactobacillus microorganisms selected from the group consisting of Lactobacillus brevis and Lactobacillus hilgadii.
(2) The production method of (1), wherein the fermentation conditions with a microorganism of the genus Lactobacillus are pH 4.5 to 7.5, 20 to 40 ° C., 20 hours to 50 days.
(3) The production method of (1) or (2), wherein the microorganism is Lactobacillus brevis or Lactobacillus hirugardi.
(4) Lactic acid fermented sake lees fermented extract containing 4,000 mg / L or more of ornithine and 60 mg / L or more of D-serine as a lactic acid fermented product derived from sake lees.
(5) Lactic acid fermented sake lees fermented extract containing less than 4,000 mg / L of ornithine and 60 mg / L or more of D-serine as a lactic acid fermented product derived from sake lees.
(6) Using sake lees as a raw material, containing 4,000 mg / L or more of ornithine and / or 60 mg / L or more of D-serine under the conditions of pH 4.5 to 7.5, 20 to 40 ° C., 20 hours to 50 days. Lactobacillus microorganisms that can produce extracts,
(7) Deposit number: Lactobacillus brevis strain deposited as NITE P-02167, and (8) Deposit number: Lactobacillus hilgardi strain deposited as NITE P-02160 or NITE P-02161,
Regarding.

According to the present invention, a lactic acid fermented sake lees fermented extract containing a functional material such as L-ornithine and / or D-amino acid can be obtained from sake lees.

In one embodiment, the present invention is obtained by a process comprising subjecting liquor lees to starch-degrading enzyme treatment and / or proteolytic enzyme treatment and fermentation treatment with yeast, followed by solid-liquid separation to remove solid components. The present invention relates to a method for producing a lactic acid fermented sake lees fermented extract, which comprises fermenting the obtained fermented sake lees extract with a microorganism of the genus Lactobacillus.

The lactic acid fermented sake lees fermented extract of the present invention is prepared from sake lees as a raw material. Sake lees are residues produced by alcoholic fermentation using rice as a raw material, and include, for example, ordinary sake lees, ginjo sake lees, and liquefied sake lees. Liquefied sake lees are sake lees prepared by preliminarily liquefying raw rice by enzyme treatment. Preferably, the sake lees used as a raw material is liquefied sake lees. The sake lees used as a raw material may or may not be sterilized by heating.

Sake lees are made into suspensions by adding water during enzyme treatment and fermentation treatment. At this time, the following enzymes, yeast, organic acids and the like may be added at the same time, or these may be added to sake lees as a suspension or an aqueous solution.

Preferably, the raw sake lees are pH adjusted prior to the enzyme treatment and / or the fermentation treatment. Preferably, the pH adjustment is done with an acid. The acid used in this case may be any acid as long as it can be used in foods, and for example, inorganic acids such as phosphoric acid and hydrochloric acid, organic acids such as acetic acid, citric acid, lactic acid and succinic acid may be used. Good. Preferably, the acid is an organic acid, more preferably lactic acid.

When adjusting the pH, it is preferable to adjust the pH to 5 or less, preferably less than pH 5, and more preferably pH 4.5 or less. Examples of preferable pH are pH 2-5, pH 2.5-5, pH 2.5-4.5, pH 3-4.8, pH 3.5-4.5 and the like.

In the production of the lactic acid fermented sake lees fermented extract of the present invention, sake lees are subjected to starch-degrading enzyme treatment and / or proteolytic enzyme treatment, and fermentation treatment with yeast. The starch-degrading enzyme treatment, the proteolytic enzyme treatment, and the fermentation treatment with yeast may be performed in any order, and two of these treatments may be performed at the same time, and the remaining one treatment may be performed before and after the treatment. Further, all of these processes may be performed at the same time. By performing all the treatments at the same time, the production time of the lactic acid fermented sake lees fermented extract can be shortened, and the process and equipment can be simplified.

The starch-degrading enzyme used in the present invention may be any starch-degrading enzyme as long as it can be used in foods. Good. Examples of the starch-degrading enzyme that can be used for producing the material of the present invention include amylases such as α-amylase and glucoamylase, glucosidases such as α-glucosidase and transglucosidase, and debranching enzymes such as pullulanase and isoamylase. These enzymes may be refined products or crude products. Those containing other enzymes such as proteolytic enzymes other than starch-degrading enzymes such as pancreatin may be used. Commercially available starch-degrading enzyme preparations can also be used in the present invention. Commercially available starch-degrading enzyme agents include Gluc 100 (manufactured by Amano Enzyme), Gluc SB (manufactured by Amano Enzyme), Gluczyme AF6 (manufactured by Amano Enzyme), Kokugen L (manufactured by Yamato Kasei Co., Ltd.), and Spitzase CP-40FG (manufactured by Yamato Kasei). Nagase ChemteX (manufactured by Nagase Chemtex), Sumiteam S (manufactured by Shin Nihon Kagaku Kogyo), Biozyme A (manufactured by Amano Enzyme), Crystase L1 (manufactured by Yamato Kasei), Coclase-G2 (manufactured by Sankyo Lifetech) .. The starch-degrading enzyme or enzyme preparation used in the production method of the present invention may be one kind or two or more kinds. These starch-degrading enzymes and commercially available starch-degrading enzyme agents are examples, and those other than the above can be appropriately selected and used depending on the type of raw material, the amount to be treated, the target product, and the like.

The proteolytic enzyme used in the present invention may be any proteolytic enzyme as long as it can be used in foods. For example, a proteolytic enzyme derived from filamentous fungi such as aspergillus, a proteolytic enzyme derived from bacteria such as Bacillus subtilis, and plant-derived papain. , Animal-derived pepsin, trypsin, etc. may be used. Examples of the proteolytic enzyme that can be used for producing the material of the present invention include peptidase and the like. These enzymes may be refined products or crude products. Those containing other enzymes such as starch degrading enzyme other than proteolytic enzyme such as pancreatin may be used. Commercially available proteolytic enzyme preparations can also be used in the present invention. Commercially available proteolytic enzyme agents include pepsin (manufactured by Wako Junyaku Co., Ltd.), Orientase 20A (manufactured by HBI), Orientase 90N (manufactured by HBI), Orientase ONS (manufactured by HBI), and Neulase F3G (Amano). Enzyme), Protease A "Amano" G (Amano Enzyme), Protease M "Amano" G (Amano Enzyme), Sumiteam AP (Shin Nihon Kagaku Kogyo), Sumiteam LP (Shin Nihon Kagaku Kogyo) Made) and so on. The proteolytic enzyme or enzyme preparation used in the production method of the present invention may be one kind or two or more kinds. These proteolytic enzymes and commercially available proteolytic enzyme agents are examples, and those other than the above can be appropriately selected and used depending on the type of raw material, the amount to be treated, the target product, and the like.

Preferably, the yeast fermentation treatment is performed before, after, or in parallel with the starch-degrading enzyme treatment and / or the proteolytic enzyme treatment. The yeast used in the present invention may be any yeast as long as it can be used in foods, for example, yeasts such as sake yeast, beer yeast, wine yeast, bread yeast, and in another classification, for example, Saccharomyces. Yeasts such as Zygosaccharomyces, Schizosaccharomyces, and Pichia may be used. The yeast used for producing the fermented sake lees extract may be one type or two or more types. These yeasts are examples, and those other than the above can be appropriately selected and used depending on the type of raw material, the amount to be treated, the target product, and the like.

After the enzyme treatment and fermentation are complete, the reaction is preferably heated to inactivate the enzyme. The heating conditions for deactivation can be appropriately adjusted by those skilled in the art, and examples thereof include heating at a temperature reached 70 ° C. It can also be used in the production of a lactic acid fermented sake lees fermented extract without heating the reaction product while retaining the enzymatic activity.

By solid-liquid separation of the obtained reaction product, the liquid portion is used as the fermented sake lees extract of the present invention. As the solid-liquid separation method, a method known to those skilled in the art can be used, and for example, centrifugation, filtration, decantation and the like can be used. The above solid-liquid separation method or a solid-liquid separation method other than the above can be appropriately selected or used in combination depending on factors such as the required amount of material and the nature of precipitation.

The fermented sake lees extract obtained by solid-liquid separation may be directly used for the next fermentation step, or may be concentrated before the next fermentation step. It may also be concentrated to an appropriate concentration during the fermentation step. A method well known to those skilled in the art can be used as a means of concentration. For example, the concentration can be performed by an evaporator or the like. Further, it may contain alcohol, or the alcohol may be removed by means well known to those skilled in the art such as evaporation. It is preferable to remove the alcohol.

It is preferable to adjust the pH of the obtained fermented sake lees extract before subjecting it to the next fermentation step. Examples of preferred pH are 4.5-7.5, 4.5-7, 5-7 and the like. A method known to those skilled in the art can be used for adjusting the pH, and for example, addition of an acid or alkali can be used.

In addition, a component that assists fermentation such as sugar may be added to the obtained fermented sake lees extract, if necessary.

The obtained fermented sake lees extract is then subjected to fermentation by a microorganism of the genus Lactobacillus. The microorganism of the genus Lactobacillus used is preferably Lactobacillus brevis or Lactobacillus hirugardi. These may be used alone or in combination of two or more. Examples of preferred Lactobacillus microorganisms are Lactobacillus brevis 376-5 strain, Lactobacillus hirugardi 376-80 strain, Lactobacillus hilgardi 301-259 strain, Lactobacillus brevis 301-282 strain, and the like.

The conditions for fermentation by microorganisms of the genus Lactobacillus can be appropriately adjusted by those skilled in the art. As more specific fermentation conditions, for example, examples of preferable temperature conditions are 20-40 ° C, 20-37 ° C, 25-40 ° C, 25-37 ° C, 25-30 ° C, 30-40 ° C, and preferred time. Examples are 20 hours or more, 20 hours to 5 days, 20 hours to 50 days, 1 day or more, 1 to 7 days, 1 to 14 days, 1 to 50 days, 2 to 14 days, 2 to 21 days, 5 to 5 15 days, 5 to 21 days, 5 to 50 days, and so on.

Lactic acid fermented sake lees fermented extract is obtained by fermentation by microorganisms of the genus Lactobacillus. The resulting lactic acid fermented sake lees fermented extract contains a high content of ornithine and / or D-serine.

The amount of ornithine in the obtained lactic acid fermented sake lees fermented extract is preferably 4,000 mg / L or more, more preferably 4,500 mg / L or more, still more preferably 5,000 mg / L or more. The D-serine content of the obtained lactic acid fermented sake lees fermented extract is preferably 60 mg / L or more, more preferably 100 mg / L or more, still more preferably 200 mg / L or more.

The obtained lactic acid fermented extract may be used as it is as a lactic acid fermented liquor lees fermented extract, or the liquid portion obtained by removing the bacterial cells from the lactic acid fermented extract may be used as the lactic acid fermented liquor lees fermented extract in the present invention. As a method for removing the bacterial cells, a method known to those skilled in the art can be used, and for example, centrifugation, filtration or the like can be used. The obtained lactic acid fermented sake lees fermented extract is subjected to a concentration or drying treatment, if desired.

In another aspect, the present invention relates to a lactic acid fermented sake lees fermented extract containing a high content of ornithine and / or D-serine.

The lactic acid fermented sake lees fermented extract of the present invention contains ornithine preferably 4,000 mg / L or more, more preferably 4,500 mg / L or more, still more preferably 5,000 mg / L or more. Alternatively, the lactic acid fermented sake lees fermented extract of the present invention contains D-amino acid, preferably D-serine, preferably 60 mg / L or more, more preferably 100 mg / L or more, still more preferably 200 mg / L or more. In certain embodiments, the lactic acid fermented sake lees fermented extract of the present invention contains both ornithine and D-serine in the above contents. In another embodiment, the lactic acid fermented extract of the present invention contains D-serine in the above content, but contains ornithine less than 4,000 mg / L, for example 2,000 mg / L or less, 1,000 mg / L or less. It is contained in a content of 500 mg / L or less.

In another embodiment, the lactic acid fermented sake lees fermented extract of the present invention contains GABA of 2,000 mg / L or more, preferably 3,000 mg / L or more. In still other embodiments, the lactic acid fermented sake lees fermented extract of the present invention contains GABA less than 2,000 mg / L, for example, less than 2,000 mg / L, 1,000 mg / L or less, 500 mg / L or less.

The lactic acid fermented sake lees fermented extract of the present invention can contain ornithine or D-serine in the above amount only by the lactic acid fermented product derived from sake lees without adding ornithine or D-serine separately. The lactic acid fermented sake lees fermented extract of the present invention may or may not have ornithine or D-serine added separately.

The lactic acid fermented sake lees fermented extract of the present invention can be expected to have functions such as improvement of liver function, promotion of growth hormone secretion, promotion of metabolism, improvement of immune function, improvement of fatigue, elimination of hangover, and improvement of endurance by the action of ornithine. In addition, the action of D-serine can be expected to improve neurological diseases such as schizophrenia, improve memory / learning ability, improve sleep, improve skin barrier function, and improve food flavor.

The lactic acid fermented sake lees fermented extract of the present invention can be used in foods, seasonings, functional foods, pharmaceuticals, cosmetics and the like. The lactic acid fermented sake lees extract of the present invention may be used as it is, or other raw materials usually used for each product may be added.

The lactic acid fermented sake lees fermented extract of the present invention can be blended into each product by a method known per se, and the blending amount and other blending components are not particularly limited. Examples of other ingredients include seasonings, acidulants, sweeteners, spices, colorants, fragrances, salts, sugars, antioxidants, vitamins, stabilizers, thickeners, carriers, excipients, lubricants, surfactants, interfaces. Activators, propellants, preservatives, chelating agents, pH adjusters, etc. may be mentioned. These other ingredients may be added at any time as long as they do not interfere with the steps described above.

The form of food, seasoning, functional food, or pharmaceutical is not particularly limited, and for example, solid (tablet, powder, granule, etc.), semi-solid (paste, etc.), liquid (solution, suspension, emulsion, etc.), Alternatively, it may be encapsulated. The type of food used is not particularly limited, and examples thereof include noodles, processed meat products, processed fish products, processed vegetable products, prepared foods, confectionery, and beverages. The type of seasoning is not particularly limited, and examples thereof include soup stock, soup, and sauce.

The form of cosmetics is not particularly limited, and for example, liquid or solid cosmetics such as cleaning cosmetics, lotions, lotions, creams, emulsions, foundations, lipsticks, eyeliners, mascaras, eye shadows, manicures, shampoos, etc. It can be a rinse, a treatment, a hair tonic, a hair styling product, a hair dye, a shaving agent, a mouthwash, a bathing agent, etc. (including non-medicinal products). From the viewpoint of the effect of fermentation products, skin (skin conditioning) cosmetics and bath salts are particularly preferable.

The present invention relates to a microorganism capable of highly producing a functional material in another embodiment. The microorganism is preferably a microorganism of the genus Lactobacillus, more preferably Lactobacillus brevis or Lactobacillus hirugardi.

The microorganism of the present invention highly produces a specific functional material. Preferably, the microorganism of the present invention highly produces D-amino acids such as ornithine or D-serine. In one embodiment, the microorganism of the present invention uses sake lees as a raw material and ornithine at 4,000 mg / L or more and / or D under the conditions of pH 4.5 to 7.5, 20 to 40 ° C., 20 hours to 50 days. -An extract containing 60 mg / L or more of serine can be produced.

In certain embodiments, high ornithine production is defined as pH: 4.5-7.5, eg 4.5-7.0; temperature: 20-40 ° C, eg 30-40 ° C; time: 20 hours-50 days. For example, the ornithine content of the lactic acid fermented sake lees fermented extract is 4,000 mg / L or more, preferably 4,500 mg / L or more, and more preferably 5,000 mg / L or more under the condition of 20 hours to 5 days. .. If the ornithine production is not high, for example, the production is low, the ornithine content in the extract is less than the above-mentioned contents under the above conditions, for example, less than 4,000 mg / L, 2,000 mg / L or less, 1,000 mg / L or less, 500 mg. / L or less, etc.

In another embodiment, high ornithine production means that the ornithine content is 3,500 mg / L or more, preferably 4,000 mg / L or more, more preferably 4,500 mg, as compared before and after the reaction under the above conditions. It means to increase by / L or more. If the ornithine production is not high, for example, if the production is low, whether the ornithine content should be increased by less than 2,000 mg / L, for example, less than 1,000 mg / L, less than 500 mg / L, or less than 100 mg / L as compared before and after the reaction. , Or will reduce the ornithine content.

In a certain embodiment, high GABA production means that the GABA content of the lactic acid fermented sake lees fermented extract is 2,000 mg / L or more, preferably 3,000 mg / L or more under the same conditions as the above-mentioned ornithine. When GABA is not high production, for example, when production is low, the GABA content in the extract is less than the above content under the above conditions, for example, less than 2,000 mg / L, 1,000 mg / L or less, 500 mg / L or less, and the like. Become.

In another embodiment, high GABA production means increasing the GABA content by 2,000 mg / L or more, preferably 3,000 mg / L or more, as compared before and after the reaction under the above conditions. If GABA is not high, for example low, the GABA content is increased by less than 2,000 mg / L, such as less than 1,000 mg / L, less than 500 mg / L, less than 100 mg / L, as compared before and after the reaction. Or it will reduce the GABA content.

In certain embodiments, high D-amino acid production refers to pH: 4.5-7.5, eg 5-7.5; temperature: 20-40 ° C, eg 25-40 ° C; time: 20 hours-50. The content of D-amino acids such as D-serine, D-aspartic acid, D-glutamic acid, D-alanine, D-valine and D-leucine of lactic acid fermented sake lees fermented extract is 60 mg / L under the condition of days, for example, 2 to 50 days. As mentioned above, it means that it is preferably 100 mg / L or more, and more preferably 200 mg / L or more. If the D-amino acid production is not high, for example, the production is low, the amino acid content in the extract is less than the above-mentioned contents under the above conditions, for example, less than 60 mg / L, 50 mg / L or less, 30 mg / L or less, 10 mg / L or less, It becomes 5 mg / L or less.

In other embodiments, high D-amino acid production refers to D-serine, D-aspartic acid, D-glutamic acid, D-alanine, D-valine and D-leucine as compared before and after the reaction under the above conditions. It means to increase the D-amino acid content such as 20 mg / L or more, preferably 50 mg / L or more, for example, 80 mg / L or more, 160 mg / L or more, 200 mg / L or more. If the D-amino acid production is not high, for example, low production, the D-amino acid content is increased by less than 20 mg / L, for example, less than 10 mg / L, less than 5 mg / L, or D-amino acid as compared before and after the reaction. The content will be reduced.

Preferably, the microorganism of the present invention highly produces one or more amino acids selected from the group consisting of D-serine, D-aspartic acid, D-glutamic acid, D-alanine, D-valine and D-leucine. More preferably, the microorganism of the present invention highly produces D-serine.

As a more specific example, as the microorganism of the present invention, (1) ornithine, D-serine, D-aspartic acid, D-glutamic acid, and D-alanine are highly produced, but GABA, D-histidine, D-proline, Microorganisms that produce less D-valine, D-isoleucine, D-leucine, D-phenylalanine; (2) Ornithine, D-serine, D-aspartic acid, D-glutamic acid, D-alanine, D-valine, D-leucine High production, but low production of GABA, D-histidine, D-proline, D-isoleucine, D-phenylalanine; (3) GABA, D-serine, D-aspartic acid, D-glutamic acid, D-alanine , D-valine, D-leucine, but less ornithine, D-histidine, D-proline, D-isoleucine, D-phenylalanine; and (4) D-serine, D-aspartic acid, Microorganisms that produce high levels of D-glutamic acid, D-alanine, D-valine, and D-leucine but low levels of ornithine, GABA, D-histidine, D-proline, D-isoleucine, and D-phenylalanine. .. Specific examples of (1) include Lactobacillus brevis 375-5 strain and the like. Specific examples of (2) include Lactobacillus hirugardi 376-80 strains. Specific examples of (3) include Lactobacillus hirugardi 301-259 strains. Specific examples of (4) include Lactobacillus brevis 301-282 strain and the like.

The above-mentioned Lactobacillus hirugardi 376-80 is a strain obtained from fermented sake lees extract, and is available at the National Institute of Technology and Evaluation Patent Microorganisms Depositary Center (NPMD) (2-5-8 Kazusakamatari, Kisarazu City, Chiba Prefecture). Deposited as Deposit Number: NITE P-02160 (Consignment Date: November 12, 2015). The above-mentioned Lactobacillus hirugardi 301-259 is a strain obtained from fermented sake lees extract, and is a patented microorganism deposit center (NPMD) of the National Institute of Technology and Evaluation (NPMD) (2-5-8 Kazusakamatari, Kisarazu City, Chiba Prefecture). ) As the deposit number: NITE P-02161 (consignment date: November 12, 2015). The above-mentioned Lactobacillus brevis 301-282 is a strain obtained from fermented sake lees extract, and is a patented microorganism deposit center (NPMD) of the National Institute of Technology and Evaluation (NPMD) (2-5-8 Kazusakamatari, Kisarazu City, Chiba Prefecture). ) As the deposit number: NITE P-02167 (consignment date: December 1, 2015).

Hereinafter, the present invention will be described in more detail and concretely with reference to Examples, but the Examples are merely for the purpose of illustration and are not necessarily intended to limit the present invention.

Example 1
Preparation of lactic acid fermented sake lees fermented extract 1
2,000 kg of liquefied sake lees and 3,600 L of water (1.8 times the amount of sake lees) were mixed, and 16.0 kg of lactic acid was added so that the pH became 4.5. In addition to this, 22.4 kg (1.0 x 10 ¹⁰ pieces / g) of solid yeast, 1.0 kg of amylase agent (Gluc 100: manufactured by Amano Enzyme) (1/2000 amount of sake lees) and a protease agent (Orientase 20A: HBI) 2.5 kg (1/800 amount of sake lees) was added, and the mixture was fermented at a constant temperature of 25 ° C. for 3 days. After the fermentation was completed, the mixture was sterilized at a temperature of 70 ° C. and pressed to obtain 4,460 L of fermented sake lees extract. A part of 2,006 L of the obtained fermented sake lees extract was concentrated to 1,038 kg, alcohol was removed, and a double concentrated solution of fermented sake lees extract was obtained. The analytical values are shown in Table 1.

The obtained fermented sake lees extract double concentrate was adjusted to pH 5.0, and after sterilization, a sterilized 40% (w / v) glucose solution was added in an amount of 1/400 to the fermented sake lees extract double concentrate. Next, the seed cultures of the lactic acid bacteria Lactobacillus brevis 376-5 strain, Lactobacillus hirugardi 376-80 strain, Lactobacillus hirugardi 301-259 strain and Lactobacillus brevis 301-282 strain were concentrated twice as much as the fermented sake lees extract. 1% was added to the solution, and the cells were allowed to stand and cultured at 35 ° C. for 2 days. The contents of glutamic acid, GABA, arginine, and ornithine in the obtained fermented lactic acid sake lees extract were measured by HPLC. A double concentrate of fermented sake lees extract without lactic acid bacteria was also measured in the same manner. The results are shown in Table 2.

Example 2
Preparation of lactic acid fermented sake lees fermented extract 2
The pH of the fermented sake lees extract double concentrate prepared in Example 1 was adjusted to 6.0, and after sterilization, a sterilized 40% (w / v) glucose solution was added in an amount of 1/40 to the fermented sake lees extract double concentrate. did. Next, the seed cultures of the lactic acid bacteria Lactobacillus brevis 376-5 strain, Lactobacillus hirugardi 376-80 strain, Lactobacillus hirugardi 301-259 strain, and Lactobacillus brevis 301-282 strain were concentrated twice as much as the fermented sake lees extract. 1% was added to the solution, and the cells were allowed to stand and cultured at 25 ° C. for 21 days. We asked Shiseido Co., Ltd. to quantify the D-amino acids in the obtained fermented lactic acid sake lees extract. We also requested the measurement of the fermented sake lees extract double concentrate without lactic acid bacteria. The results are shown in Table 3.

From the results of Example 1 and Example 2, the lactic acid fermented sake lees fermented extract obtained by fermenting the sake lees fermented extract double concentrate in each strain contains a high amount of D-amino acids such as ornithine or D-serine as follows. It became clear to do. The lactic acid fermented sake lees fermented extract obtained has a high content of ornithine, D-serine, D-aspartic acid, D-glutamic acid, and D-alanine in the Lactobacillus brevis 376-5 strain. The Lactobacillus hirugardi 376-80 strain is high in ornithine, D-serine, D-aspartic acid, D-glutamic acid, D-alanine, D-valine and D-leucine. The Lactobacillus hirugardi 301-259 strain is high in GABA, D-serine, D-aspartic acid, D-glutamic acid, D-alanine, D-valine and D-leucine. The Lactobacillus brevis 301-282 strain has a high content of D-serine, D-aspartic acid, D-glutamic acid, D-alanine, D-valine, and D-leucine.

According to the present invention, a lactic acid fermented sake lees fermented extract containing a functional material can be obtained. The lactic acid fermented sake lees extract of the present invention can be used in the fields of foods, seasonings, functional foods, pharmaceuticals, cosmetics and the like.

Claims (6)

Lactobacillus brevis and fermented liquor extract obtained by subjecting liquor to starch-degrading enzyme treatment and / or proteolytic enzyme treatment and fermentation treatment with yeast, and then performing solid-liquid separation to remove solid components. A method for producing a lactic acid fermented liquor lees fermented extract, which comprises fermenting with one or more Lactobacillus microorganisms selected from the group consisting of Lactobacillus hirugardi. The production method according to claim 1, wherein the fermentation conditions with a microorganism of the genus Lactobacillus are pH 4.5 to 7.5, 20 to 40 ° C., 20 hours to 50 days. The production method according to claim 1 or 2, wherein the microorganism is Lactobacillus brevis or Lactobacillus hirugardi. Using sake lees as a raw material, an extract containing 4,000 mg / L or more of ornithine and / or 60 mg / L or more of D-serine is produced under the conditions of pH 4.5 to 7.5, 20 to 40 ° C., and 20 hours to 50 days. A microorganism of the genus Lactobacillus consisting of Lactobacillus brevis or Lactobacillus hirugardi. Accession number: Lactobacillus brevis strain deposited as NITE P-02167. Accession number: Lactobacillus hirugardi strain deposited as NITE P-02160 or NITE P-02161.