Fermentation production method

Abstract

To provide a method and so on for producing a novel fermented product.SOLUTION: A fermented carrot is produced through enzymatic treatment with lactase and fermentation treatment with Lactobacillus lactic acid bacteria.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to a method for producing a fermented product. [Background technology]

[0002] Ginseng (medicinal ginseng), including Panax ginseng (CA Meyer), has been used as a medicinal herb since ancient times, mainly in China, the Korean Peninsula, and Japan. Its useful components are said to be saponins (ginseng saponins) with a dalaman skeleton called ginsenosides, and research has revealed that they have various physiological effects, such as antioxidant, antidiabetic, antiobesity, antihypertensive, and anticancer effects (Non-Patent Document 1).

[0003] Ginseng saponins contain various ginsenosides. Among them, protopanaxadiol, such as ginsenosides Rb1, Rb2, Rc, and Rd, is thought to be metabolically converted by intestinal bacteria to 20(S)-protopanaxadiol 20-O-β-d-glucopyranoside (also known as M1 or compound K), which then passes through the intestinal wall and enters the bloodstream, exerting the physiological effects described above. Regarding M1, fermented ginseng contains M1, and it has been reported in tests on mice and humans that stress is reduced compared to before fermentation (Patent Document 1) and that the sleep-improving effect is increased (Non-Patent Document 2).

[0004] Fermented ginseng is obtained by fermenting ginseng with a microorganism, and Patent Document 1 discloses a fermentation method using, as the microorganism, a microorganism that produces at least one enzyme selected from the group consisting of β-glucosidase, α-arabinosidase, and α-rhamnosidase, which is preferably a microorganism that can be added to foods, specifically, Lactobacillus casei Hasegawa strain (FERM BP-10123). [Prior art documents] [Patent documents]

[0005] [Patent Document 1] Patent No. 5204771 [Non-patent literature]

[0006] [Non-Patent Document 1] Journal of Ginseng Research 37, 261-268(2013) [Non-patent document 2] SLEEP 32, 413-421(2009) Summary of the Invention [Problem to be solved by the invention]

[0007] An object of the present invention is to provide a novel method for producing a fermented product.

[0008] Another object of the present invention is to provide a method for producing a fermentation product that can efficiently produce M1.

[0009] Another object of the present invention is to provide a novel fermented product.

[0010] Another object of the present invention is to provide a novel microorganism. [Means for solving the problem]

[0011] As a result of extensive research, the present inventors have discovered that a novel method for producing a fermented product can be provided by combining an enzyme treatment with a fermentation treatment using Lactobacillus lactic acid bacteria (e.g., Lactobacillus casei Hasegawa strain), and that, in particular, the use of a specific enzyme (i.e., lactase) in the enzyme treatment can efficiently produce M1, thereby completing the present invention.

[0012] That is, the present invention relates to the following inventions. [1] A method for producing fermented carrots through enzymatic treatment with lactase (galactosidase) and fermentation treatment with Lactobacillus lactic acid bacteria (lactic acid bacteria belonging to the Lactobacillus genus). [2] The method according to [1], wherein the ginseng comprises at least one species selected from the group consisting of Panax ginseng, Panax notoginseng, American ginseng, Bamboo ginseng, Himalayan ginseng, and Vietnamese ginseng. [3] The method according to [1] or [2], wherein the enzyme treatment is carried out at an enzyme concentration of 0.01 to 20 w / v % at 30 to 60°C for 3 to 72 hours. [4] The method according to any one of [1] to [3], wherein the Lactobacillus lactic acid bacteria comprises at least one species selected from Lactobacillus casei and Lactobacillus brevis. [5] The method according to any one of [1] to [4], wherein the Lactobacillus lactic acid bacteria comprises at least one species selected from the group consisting of Lactobacillus casei Hasegawa strain (FERM BP-10123), Lactobacillus casei NBRC15883 strain, Lactobacillus brevis DNBL1871 strain (accession number NITE P-02661), and Lactobacillus brevis DNBL1889 strain (accession number NITE AP-03235, accession number NITE P-03235). [6] The method according to any one of [1] to [5], wherein the Lactobacillus lactic acid bacteria includes Lactobacillus brevis DNBL1889 strain (accession number NITE AP-03235, accession number NITE P-03235). [7] The method according to any one of [1] to [6], wherein the enzyme treatment is carried out simultaneously with or before the fermentation treatment. [8] The method according to any one of [1] to [6], wherein the fermentation treatment is carried out simultaneously with or before the enzyme treatment. [9] The method according to any one of [1] to [8], wherein the fermented product contains M1.

[10] A fermented carrot product obtained by the method according to any one of [1] to [9].

[11] A fermented carrot product containing M1 in a proportion of 0.75% by mass or more (e.g., 0.8% by mass or more, 0.85% by mass or more, 0.9% by mass or more).

[12] A fermented carrot product containing M1, wherein the M3 / M1 area ratio in HPLC is 0.4 or less.

[13] The fermented product according to

[11] or

[12] , which does not contain ginsenoside Rb1.

[14] The fermented product according to

[12] or

[13] , containing M1 at a rate of 0.4% by mass or more.

[15] A composition comprising the fermented product according to any one of

[10] to

[14] .

[16] Lactobacillus brevis strain DNBL1889 (accession number NITE AP-03235, accession number NITE P-03235). [Effects of the Invention]

[0013] According to the present invention, a novel method for producing a fermented product can be provided. Such a method combines a specific enzyme treatment with a fermentation treatment using specific lactic acid bacteria, and can efficiently carry out fermentation (e.g., metabolism or hydrolysis) [e.g., can efficiently produce M1 (can produce a fermented product containing a high proportion of M1)].

[0014] Another aspect of the present invention provides a novel fermented product, which can be produced, for example, by the above-described method, i.e., a method that combines a specific enzyme treatment with a fermentation treatment using a specific lactic acid bacterium, and which can have characteristics such as a high content of M1.

[0015] Another aspect of the present invention provides a novel microorganism that can be used in the fermentation of carrots, and is particularly suitable as a lactic acid bacterium in the above-mentioned method (i.e., a method that combines a specific enzyme treatment with a fermentation treatment using a specific lactic acid bacterium). DETAILED DESCRIPTION OF THE INVENTION

[0016] [Manufacturing method] In the present invention, a fermented carrot product is produced through an enzymatic treatment with lactase (galactosidase) and a fermentation treatment with Lactobacillus lactic acid bacteria (lactic acid bacteria belonging to the genus Lactobacillus).

[0017] As the ginseng (medicinal ginseng), for example, Araliaceae ginseng (Araliaceae ginseng) can be used. Such ginseng may generally be capable of producing M1 by fermentation.

[0018] Representative examples of ginseng (Araliaceae ginseng) include, for example, Korean ginseng (Panax CA Meyer), Panax notoginseng Burk., American ginseng (Panax quinquefolium L.), bamboo-jointed ginseng (Panax japonicus CA Meyer), Himalayan ginseng (Panax Pseudo-ginseng Qall. Subsp. Himalaicus Hara), and Vietnamese ginseng (Panax Vuetnamensis Ha et Grushv.).

[0019] Carrots may be used alone or in combination of two or more types.

[0020] Carrots may be natural (unprocessed) or processed. Examples of processed products include dried carrots, cut carrots, crushed carrots, extracts, pastes, etc. Conventional methods can be used for drying, cutting, crushing, extracting, and pasting.

[0021] The carrot may be any of dried carrot, white carrot, red carrot, etc.

[0022] The size of the carrots (processed products) is not particularly limited, but may be, for example, an average major axis of 0.2 mm or less. The carrots may be commercially available.

[0023] The part of the carrot is not particularly limited, and any part can be used. Examples include roots, stems, leaves, flower buds, fruits, and the whole plant. One or more of these can be used. Roots are preferred, and lateral roots, taproots, and the like are more preferred.

[0024] In the enzymatic treatment, lactase [galactosidase (β-galactosidase)] is used as the enzyme.

[0025] The enzyme may include lactase, and may also include other enzymes.

[0026] The enzyme is not particularly limited in terms of the microorganism from which it is derived. For example, lactase may be derived from a fungus (e.g., the genus Aspergillus [e.g., Aspergillus oryzae (Aspergillus oryzae)], or a yeast (e.g., the genus Kluveromyces [e.g., Kluveromyces lactis]).

[0027] The enzyme may be a commercially available product. For example, lactase is available from Amano Enzyme Co., Ltd. as Lactase F "Amano."

[0028] The enzymatic treatment method is not particularly limited, and is usually carried out by contacting the carrot with an enzyme.

[0029] The enzyme treatment may be carried out in the presence of a medium (reaction medium) (for example, in water).

[0030] In the enzymatic treatment, the amount (proportion) of the enzyme (lactase) used may be, for example, about 0.001 part by mass or more, preferably 0.005 part by mass or more, and more preferably 0.01 part by mass or more, per 1 part by mass of carrot.

[0031] The amount (ratio) of the enzyme (lactase) used may be, for example, about 0.2 parts by mass or less, preferably 0.1 parts by mass or less, and more preferably 0.08 parts by mass or less, per 1 part by mass of carrot.

[0032] In the present invention, it is possible to efficiently obtain a fermented product even when a small amount of enzyme (lactase) is used.

[0033] In enzyme treatment (e.g., enzyme treatment in a medium such as water), the ratio of the enzyme (or concentration, e.g., the ratio to the medium such as water) may be, for example, 0.01 w / v% or more (e.g., 0.05 w / v% or more), preferably 0.1 w / v% or more (e.g., 0.5 w / v% or more), and more preferably 1 w / v% or more. In the enzyme treatment, the upper limit of the enzyme proportion (concentration) may be, for example, 50 w / v%, 40 w / v%, 30 w / v%, 20 w / v%, 15 w / v%, 10 w / v%, 8 w / v%, 5 w / v%, etc.

[0034] In enzyme treatment (e.g., enzyme treatment in a medium such as water), the proportion of carrot (or concentration, e.g., the proportion relative to the medium such as water) may be, for example, 1 w / v% or more (e.g., 2 w / v% or more), preferably 5 w / v% or more (e.g., 10 w / v% or more), more preferably 15 w / v% or more, or may be 50 w / v% or less, 40 w / v% or less, 30 w / v% or less, etc.

[0035] In the enzyme treatment, the temperature (enzyme treatment temperature) may be, for example, about 15 to 80°C (eg, 20 to 70°C), preferably about 30 to 60°C, and more preferably about 40 to 55°C.

[0036] In the enzyme treatment, the time (enzyme treatment time) may be, for example, 1 hour or more, preferably 3 hours or more, and more preferably 8 hours or more (e.g., 15 hours or more), or may be 10 days or less, 8 days or less, 5 days or less, 3 days (72 hours) or less, 48 ​​hours or less, 24 hours or less, etc.

[0037] The fermentation treatment is carried out using (in the presence of) Lactobacillus lactic acid bacteria (lactic acid bacteria belonging to the genus Lactobacillus) as the microorganism.

[0038] Examples of Lactobacillus lactic acid bacteria include Lactobacillus acidophilus. Lactobacillus acidphilus, Lactobacillus gasseri, Lactobacillus mali, Lactobacillus plantarum, Lactobacillus buchneri, Lactobacillus casei, Lactobacillus johnsonii, Lactobacillus gallinarum, Lactobacillus amylovorus, Lactobacillus brevis, Lactobacillus rhamnosus, Lactobacillus kefir, Lactobacillus paracasei, Lactobacillus crispatus, Lactobacillus pentosus, etc.

[0039] The strain of each bacterial species is not particularly limited.

[0040] For example, examples of Lactobacillus casei (strains) include Lactobacillus casei Hasegawa strain (FERM BP-10123), Lactobacillus casei ATCC393 strain, Lactobacillus casei NBRC15883 strain, and the like.

[0041] Furthermore, for example, examples of Lactobacillus brevis (strains) that can be used include Lactobacillus brevis ATCC14869 strain, Lactobacillus brevis JCM1559 strain, Lactobacillus brevis NBRC12005 strain, Lactobacillus brevis DNBL1867 strain, Lactobacillus brevis DNBL1868 strain, Lactobacillus brevis DNBL1869 strain, Lactobacillus brevis DNBL1870 strain, Lactobacillus brevis DNBL1871 strain (accession number NITE P-02661), and Lactobacillus brevis DNBL1889 strain (accession number NITE AP-03235, accession number NITE P-03235).

[0042] The microorganisms may be deposited. For example, Lactobacillus casei Hasegawa strain (FERM BP-10123) was deposited at the National Institute of Advanced Industrial Science and Technology (AIST), Patent Organism Depositary (Address: Central 6, 1-1-1 Higashi, Tsukuba, Ibaraki, Japan, Postal Code 305-8566) on August 11, 2003 (Deposit Date), under Accession Number FERM BP-10123. Lactobacillus brevis DNBL1871 strain (Accession Number NITE P-02661) was deposited at the National Institute of Technology and Evaluation (NPMD), Patent Microorganism Depositary (Address: Room 122, 2-5-8 Kazusa Kamatari, Kisarazu, Chiba, Postal Code 292-0818) on March 6, 2018 (Deposit Date), under Accession Number NITE P-02661. The Lactobacillus brevis strain DNBL1889 (accession number NITE AP-03235, accession number NITE P-03235) was deposited under the accession number NITE AP-03235 on June 22, 2020 (date of receipt) at the National Institute of Technology and Evaluation, Patent Microorganisms Depositary (NPMD) (address: Room 122, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, 292-0818 Japan).

[0043] Among these Lactobacillus lactic acid bacteria, Lactobacillus casei (for example, Lactobacillus casei Hasegawa strain) and Lactobacillus brevis (for example, Lactobacillus brevis DNBL1889 strain, Lactobacillus brevis DNBL1871 strain) may be preferably used. In particular, Lactobacillus brevis strain DNBL1889, Lactobacillus brevis strain DNBL1871, etc., when combined with enzyme treatment (and further combined with other Lactobacillus lactic acid bacteria (e.g., Lactobacillus casei Hasegawa strain)), can dramatically improve the efficiency of fermentation (metabolism or hydrolysis).

[0044] The Lactobacillus brevis strain DNBL1889 is a novel microorganism (lactic acid bacterium). Therefore, the present invention encompasses this novel microorganism. As described above, this novel microorganism is useful in combination with enzyme treatment. It also appears to possess properties that make it useful for the fermentation (metabolism or hydrolysis) of ginsenosides, particularly ginsenoside F2.

[0045] Lactobacillus lactic acid bacteria (species, strains) may be used alone or in combination of two or more kinds.

[0046] As long as the microorganisms (fermentation treatment) contain (use) Lactobacillus lactic acid bacteria, other microorganisms may also be contained (used).

[0047] The fermentation treatment can usually be carried out by bringing carrots into contact with microorganisms (lactobacillus lactic acid bacteria).

[0048] The fermentation process (fermentation method) may be carried out in a medium (in a medium). For example, fermentation may be carried out in (via) a medium. When using such a medium, for example, fermentation can be carried out by inoculating a medium (culture medium) containing carrots with microorganisms.

[0049] The medium may be sterilized (treated) before being inoculated with the microorganism. The sterilization method is not particularly limited, and examples thereof include heat sterilization, high-pressure steam sterilization, and filtration sterilization.

[0050] The medium (culture medium) is not particularly limited, but may be, for example, a medium containing a carbon source, a nitrogen source, a mineral source, etc. that are commonly used in culturing microorganisms, and may be either a natural medium or a synthetic medium, etc. Preferably, a liquid medium may be used.

[0051] The nitrogen source is not particularly limited, and examples of inorganic nitrogen sources include ammonia and ammonium salts, while examples of organic nitrogen sources include peptone, polypeptone, urea, amino acids, proteins, and peptides such as soybean peptides. The nitrogen source is preferably peptone, polypeptone, or peptide. Furthermore, examples of mineral sources are not particularly limited, and include yeast extract, meat extract, and, in addition, potassium monohydrogen phosphate, magnesium sulfate, and the like, which contain K, P, Mg, S, and the like. These nitrogen and mineral sources can be used alone or in combination of two or more. The concentration of the nitrogen source in the medium (culture medium) is not particularly limited as long as it is a normal concentration at which the microorganism can grow. The concentration of the nitrogen source at the start of cultivation is usually preferably about 0.05 to 10% by weight, more preferably about 0.1 to 5% by weight.

[0052] In addition to the nitrogen source and mineral source, the medium (culture medium) may further contain a carbon source, inorganic substances, pH buffers, etc. Examples of inorganic substances include, but are not limited to, ammonium sulfate, potassium phosphate, magnesium chloride, salt, iron, manganese, molybdenum, various vitamins, etc. These can be used alone or in combination of two or more. Examples of pH buffers include, but are not limited to, calcium carbonate, etc.

[0053] In fermentation treatment (e.g., fermentation treatment in a medium (culture medium)), the proportion of carrot (or concentration, e.g., the proportion relative to the medium (fermentation liquid)) is not particularly limited and can be selected appropriately depending on the type, shape, drying state, culture conditions, etc., and may be selected, for example, from a range of about 0.1 w / v% or more, 1 w / v% or more (e.g., 2 w / v% or more), preferably 5 w / v% or more (e.g., 10 w / v% or more), more preferably 15 w / v% or more, or may be 50 w / v% or less, 40 w / v% or less, 30 w / v% or less, etc.

[0054] Furthermore, in the fermentation treatment, the proportion of carrots may be, for example, 0.1 to 100 parts by mass (e.g., 0.5 to 80 parts by mass), preferably 1 to 50 parts by mass (e.g., 2 to 30 parts by mass), and more preferably 3 to 20 parts by mass (e.g., 5 to 18 parts by mass, 10 to 15 parts by mass, etc.) per 100 parts by mass of the medium (medium not containing carrots, total amount of medium other than carrots).

[0055] The carrots may be dried (for example, dried at an internal temperature of about 100-180°C for a specified time (about 1-6 hours)), and if not dried, the amount (ratio) used may be converted into the amount of dried carrots (for example, if undried carrots are used, the amount may be converted into the amount of dried carrots used).

[0056] In the fermentation treatment, the pH of the system [or medium (culture medium)] may be, for example, about 3 to 7, and preferably about 5 to 6.5. The pH may be controlled or adjusted using an acid or alkali.

[0057] In the fermentation treatment, the temperature (fermentation treatment temperature) may be about 5°C or higher (e.g., 8°C or higher), for example, 10°C or higher (e.g., 12 to 60°C), preferably 15°C or higher (e.g., 18 to 55°C), more preferably 20°C or higher (e.g., 22 to 50°C), or may be 25 to 40°C (e.g., 25 to 37°C, 28 to 33°C), etc.

[0058] The time (fermentation treatment time) for fermentation can be appropriately selected depending on the fermentation conditions [e.g., the composition of the system (e.g., the amount of microorganisms), temperature, etc.], and may be, for example, 6 hours or more (e.g., 12 hours or more, 1 day or more), 2 days or more, 3 days or more, 5 days or more, 7 days or more, etc. The upper limit of the fermentation time is not particularly limited, and may be 3 months, 2 months, 1 month, 25 days, 21 days, 14 days, 10 days, etc.

[0059] As described above, in the present invention, the enzyme treatment and the fermentation treatment are combined, but these treatments may be carried out simultaneously (or in parallel), or one treatment may be carried out after the other treatment.

[0060] The other treatment may be carried out after separating (the processed product of) one treatment, or may be carried out after one treatment in the same system without separation.

[0061] In particular, in the present invention, it is preferred that the enzyme treatment be carried out simultaneously with or before the fermentation treatment, and it is particularly preferred that the enzyme treatment be carried out before (prior to) the fermentation treatment. By combining the two treatments in this way, it becomes easier to obtain a fermented product (for example, a fermented product containing a high content of M1) more efficiently. On the other hand, in the present invention, from the viewpoints of easily reducing thermal history, easily reducing damage to materials, and improving the efficiency of the manufacturing process, it is preferable to carry out the fermentation treatment simultaneously with or before the enzyme treatment, and in particular, before (prior to) the fermentation treatment.

[0062] The method of the present invention may include (or undergo) an enzyme treatment (enzyme treatment step) and a fermentation treatment (fermentation treatment step), and may also include (or undergo) other treatments (steps) as necessary. For example, before, during, and / or after these treatments, treatments such as filtration, centrifugation, concentration, ultrafiltration, lyophilization, powderization, and fractionation may be carried out as desired. As a specific example, sterilization (heat treatment, etc.) may be carried out after the fermentation treatment, which can kill the microorganisms used in the fermentation treatment. It should be noted that such sterilization treatment may be omitted (or may be substituted for a step after the fermentation step) depending on the step after the fermentation treatment. For example, when enzyme treatment is carried out in parallel with or after fermentation treatment, the enzyme treatment may be carried out without sterilization treatment (heat treatment, etc.).

[0063] [Fermented products, compositions, and their uses] A fermented product (fermented carrot product) is obtained as described above. Such fermentations typically contain M1 [20(S)-protopanaxadiol 20-O-β-d-glucopyranoside, compound K].

[0064] In such fermented products, M1 is often efficiently produced by the above-mentioned method. For example, when the amount of M1 produced (content in the fermented product) is set to 1 when only fermentation treatment [fermentation treatment using Lactobacillus lactic acid bacteria (e.g., Lactobacillus casei Hasegawa strain (FERM BP-10123)] is performed, the amount can be made to be greater than 1, preferably 1.1 or more, more preferably 1.2 or more, and particularly 1.3 or more (e.g., 1.4 or more, 1.5 or more, 1.6 or more, 1.7 or more, 1.8 or more, 2 or more), etc.

[0065] The content of M1 in the fermented product (content in the fermented product) is not particularly limited, but may be, for example, 0.1% by mass or more (e.g., 0.2% by mass or more), preferably 0.3% by mass or more (e.g., 0.35% by mass or more), more preferably 0.4% by mass or more (e.g., 0.45% by mass or more), or 0.5% by mass or more (e.g., 0.6% by mass or more, 0.7% by mass or more, 0.75% by mass or more, 0.8% by mass or more, 0.9% by mass or more, 1% by mass or more, 1.1% by mass or more, 1.2% by mass or more, 1.3% by mass or more, 1.4% by mass or more), etc.

[0066] In addition, the fermented product may contain components other than M1 {e.g., ginsenoside Rb2, ginsenoside Rc, ginsenoside Rd, ginsenoside Re, ginsenoside Rg1, gypenoside XVII, M2 [20(S)-protopanaxadiol 20-O-[β-L-arabinopyranosyl(1→6)-β-D-glucopyranoside], M3 [20(S)-protopanaxadiol 20-O-[β-L-arabinofuranosyl(1→6)-β-D-glucopyranoside], etc.]}.

[0067] In the fermented product, the ratio of M3 to M1 (M3 / M1), as an area ratio (peak area ratio) in HPLC, can be selected from a range of about 0.5 or less (e.g., 0.48 or less), and may be 0.45 or less (e.g., 0.43 or less), preferably 0.42 or less (e.g., 0.4 or less, 0.38 or less, 0.35 or less, 0.32 or less, 0.3 or less, 0.28 or less), more preferably 0.25 or less (e.g., 0.24 or less, 0.22 or less, 0.21 or less), or may be 0.2 or less (e.g., 0.19 or less, 0.18 or less, 0.16 or less, 0.15 or less, etc.).

[0068] The fermentation product may not contain M3 (or may contain M1, and the M3 / M1 area ratio in HPLC may be 0).

[0069] The fermented product may also contain ginsenoside Rb1, but is often low in ginsenoside Rb1, and in particular may not contain ginsenoside Rb1 (or may contain M1, and the ginsenoside Rb1 / M1 value in HPLC area ratio may be 0).

[0070] In the present invention, a fermented product containing relatively little M3 and ginsenoside Rb1 is easily obtained.

[0071] The presence or absence and proportion of each component in such a fermented product may be confirmed, for example, by HPLC (HPLC analysis). Furthermore, "not containing" may also mean, for example, that it is not detected (no peak is detected) in HPLC (it is at the detection limit) (the same applies hereinafter).

[0072] The present invention also includes such fermented products. Note that the fermented products may or may not be obtained by the above-mentioned method (or may be obtained by a different method).

[0073] The fermented product of the present invention may be separated as is or a portion thereof (e.g., a fraction containing M1). The fermented product may also contain components used in the above-mentioned methods (e.g., enzymes, lactic acid bacteria, medium components, etc.).

[0074] The fermented product of the present invention may constitute a composition, and therefore the present invention also includes a composition containing the fermented product.

[0075] The fermented product (or composition) of the present invention can be used for various purposes, such as the prevention, amelioration, and / or treatment of various diseases or symptoms. Specifically, the fermented product (or composition) of the present invention may be used for, for example, antitumor, anti-stress, diabetes prevention (amelioration, treatment), anti-obesity, hair growth, anti-amnesia, hepatoprotective effect, anti-arteriosclerosis, anti-aging, antihyperlipidemic, antithrombotic, blood pressure lowering effect, immune function improvement, blood glucose level lowering effect, analgesic effect, cardiotonic effect, anti-inflammatory effect, peripheral blood flow improving effect, hemostatic effect, antioxidant effect, anti-muscle atrophy effect, bone mass increasing effect, anti-infectious disease (anti-influenza, etc.), neuroprotective effect, anti-fatigue, skin beautifying effect, skin whitening, etc.

[0076] The fermented product (or composition) of the present invention may be contained in (compounded into) a food or drink, or may be in any suitable form. For example, the fermented product (or composition) of the present invention may be formulated into oral or parenteral preparations such as tablets, powders, granules, capsules, liquids, emulsions, elixirs, suspensions, syrups, lozenges, inhalants, suppositories, injections, ointments, eye ointments, eye drops, nasal drops, ear drops, poultices, lotions, and the like.

[0077] When the fermented product (or composition) of the present invention is ingested (administered), the conditions for ingestion (administration) can be appropriately selected depending on the form, purpose of administration, type, age, body weight, and symptoms of the recipient, but for example, the effective human dose of M1 may be set to about 0.01 to 100 mg / kg (e.g., 1 to 50 mg / kg) per day. Administration may be performed once or in several divided doses within the desired dosage range within a day. The administration period is also optional.

[0078] The subjects to be inoculated (administered) with the fermented product (or composition) of the present invention may be humans or non-human animals (e.g., mammals such as dogs, cats, rabbits, cows, horses, goats, monkeys, and mice). Non-human animals may also be pets, livestock, or laboratory animals (e.g., mice, rats, guinea pigs, monkeys, etc.).

[0079] The fermented product (or composition) of the present invention may contain other components such as carriers, bases, additives, etc. depending on the purpose of use and form (e.g., use form, formulation form), etc. Specifically, it may contain, for example, excipients, binders, lubricants, colorants, flavorings, and, if necessary, stabilizers, emulsifiers, absorption enhancers, surfactants, pH adjusters, preservatives, antioxidants, etc.

[0080] The content of the fermented product in the composition can be selected appropriately depending on the purpose of use, form, etc., and may be, for example, 0.01% by mass or more (e.g., 0.05% by mass or more, 0.1% by mass or more, 0.3% by mass or more, 0.5% by mass or more, 1% by mass or more, 3% by mass or more, 5% by mass or more, 10% by mass or more, etc.), or less than 100% by mass (e.g., 99% by mass or less, 95% by mass or less, 90% by mass or less, 80% by mass or less, 50% by mass or less, 30% by mass or less, 20% by mass or less, 10% by mass or less, 5% by mass or less, 3% by mass or less, 1% by mass or less, etc.). [Example]

[0081] The present invention will be described in detail below with reference to examples, but the present invention is not limited to these examples.

[0082] The lactic acid bacteria and enzymes used in the examples are as follows: Lactobacillus casei Hasegawa strain (accession number: FERM BP-10123, hereinafter referred to as A221 strain) Lactobacillus casei NBRC15883 strain (also known as JCM1134T, hereinafter referred to as strain 15883) Lactobacillus brevis strain DNBL1889 (accession number NITE AP-03235, accession number NITE P-03235, hereinafter referred to as DNBL1889 strain) Lactobacillus brevis strain DNBL1871 (accession number NITE P-02661, hereinafter referred to as DNBL1871 strain)

[0083] Lactase: Lactase F (Manufacturer: Amano Enzyme Co., Ltd.) Cellulase: Y-NC (Manufacturer: Yakult Industries Co., Ltd.) β-Glucosidase: Aromasase (Manufacturer: Amano Enzyme Co., Ltd.)

[0084] Test Example 1 (1) Each lactic acid bacterium was inoculated into 6 mL of a general lactic acid bacterium inoculation medium (manufactured by Nissui Pharmaceutical Co., Ltd.) and pre-cultured overnight at 28°C. (2) 6.5 g of ginseng powder was added to 40 mL of distilled water and heated at 105°C for 5 minutes. (3) After cooling to below 50°C, 400 mg of each enzyme was added, and the mixture was shaken at 55°C and 100 rpm to carry out the enzyme reaction for 17 hours (when only enzyme treatment was carried out, i.e., when only lactic acid bacteria fermentation treatment was carried out, this step (3) (and further heating at 105°C for 5 minutes in step (2)) was not carried out). (4) After the reaction was completed, 0.5 g of yeast extract, 0.25 g of soybean peptone, and 0.5 g of calcium carbonate were added and mixed, and this was used as a fermentation medium and heat sterilized at 121°C for 15 minutes (only when lactic acid bacteria treatment was performed, i.e., when only enzyme treatment was performed, this step (4) and the step (5) described below were not performed). (5) 1.5 mL of each lactic acid bacteria preculture solution was added to the fermentation medium and allowed to stand at 28°C. (6) After 6 days, heat treatment (110 °C for 20 minutes) was performed to kill the lactic acid bacteria. Then, the fermentation medium was shaken well, and 0.8 g of each was collected. The same amount of ethanol was added thereto, and after stirring, centrifugation (10,000 rpm, 5 minutes) was performed to obtain a supernatant. 1 mL of this supernatant was added to a Sep-Pak C18 cartridge (manufactured by Waters) for adsorption, washed with distilled water and a 40% aqueous methanol solution, and then eluted with methanol (when only enzymatic treatment was performed, that is, when lactic acid bacteria fermentation treatment was not performed, the enzymatic treatment solution obtained in step (3) was used instead of the fermentation medium). (7) 10 μL of the eluted methanol solution was taken and analyzed for M1, M3, and Rb1 by high performance liquid chromatography (HPLC) under the following conditions. The relative value with respect to the peak area of M1 when only lactic acid bacteria fermentation treatment by strain A221 was performed without enzymatic treatment (at the beginning of the following table, that is, when the enzyme used in step (3) was "not used" and the lactic acid bacteria used in step (5) was "A221", the value when the peak area of M1 was set to 1), the M3 / M1 ratio (only when M1 could be detected), and the ginsenoside Rb1 / M1 ratio (only when M1 could be detected) were quantified.

[0085] <HPLC analysis conditions 1 (analysis of M1 and M3)> Column: YMC-Pack ODS-A (250 x 4.6 mm) Analysis temperature: 40 °C Eluent: 60% (V / V) acetonitrile Flow rate: 1 mL / min Detection: 203 nm

[0086] <HPLC analysis conditions 2 (analysis of Rb1 and M!> Column: YMC-Pack ODS-A (250 x 4.6 mm) Analysis temperature: 4(TC Eluent: 0 - 5 minutes: 35% (V / V) acetonitrile 5 - 35 minutes: Gradient of 35% (V / V) → 60% (V / V) acetonitrile 35 - 50 minutes: 60% (V / V) acetonitrile Flow rate: 1 mL / min Detection: 203 nm

[0087] The results are shown in the table below.

[0088] [Table 1]

[0089] Test Example 2 In "(7)" of Test Example 1, each step was carried out in the same manner as in Test Example 1, except that the relative value to the peak area of ​​M1 when only lactic acid bacteria fermentation treatment with the A221 strain was performed without enzyme treatment was quantified instead of the relative value to the peak area of ​​M1 when only lactic acid bacteria fermentation treatment with the A221 strain was performed without enzyme treatment (the value at the beginning of the table below, i.e., when the peak area of ​​M1 when the enzyme used in step (3) was "not used" and the lactic acid bacteria used in step (5) was "A221 and NBRC15883" was set to 1).

[0090] The results are shown in the table below.

[0091] [Table 2]

[0092] Test Example 3 (1) Each lactic acid bacterium was inoculated into 6 mL of a general lactic acid bacterium inoculation medium (manufactured by Nissui Pharmaceutical Co., Ltd.) and pre-cultured overnight at 28°C. (2) 13.0 g of ginseng powder was added to 40 mL of distilled water and heated at 105°C for 5 minutes. (3) After cooling to below 50°C, 400 mg of enzyme was added, and the mixture was shaken at 55°C and 100 rpm to carry out the enzyme reaction for 17 hours (when only enzyme treatment was carried out, i.e., when only lactic acid bacteria fermentation treatment was carried out, this step (3) (and further heating at 105°C for 5 minutes in step (2)) was not carried out). (4) After the reaction was completed, 1.0 g of yeast extract, 0.5 g of soybean peptone, and 1.0 g of calcium carbonate were added and mixed, and the mixture was used as a fermentation medium and sterilized by heating at 121°C for 15 minutes. (5) 3.0 mL of each lactic acid bacteria preculture solution was added to the fermentation medium and allowed to stand at 28°C. (6) After 10 days, the fermentation medium was sterilized by heating at 121°C for 15 minutes and then cooled to room temperature. (7) The fermentation medium (6) was frozen in a freezer at −20° C. and then freeze-dried for 5 days to obtain a powder. (8) 20 mg of the obtained powder was weighed out, 400 μL of 50% ethanol aqueous solution was added, and after stirring, the mixture was centrifuged (10,000 rpm for 5 minutes). 10 μL of the supernatant was analyzed under HPLC analysis condition 1 described in Test Example 1, and the M1 content [W / W (%) (mass %)] in the powder was determined. In addition, M1, M3, and Rb1 were analyzed in the same manner as in Test Example 1, and the M3 / M1 ratio (only those in which M1 was detected) and the ginsenoside Rb1 / M1 ratio (only those in which M1 was detected) were quantified.

[0093] The results are shown in the table below.

[0094] [Table 3]

[0095] Test Example 4 (1) Each lactic acid bacterium was inoculated into 6 mL of a general lactic acid bacterium inoculation medium (manufactured by Nissui Pharmaceutical Co., Ltd.) and pre-cultured overnight at 28°C. (2) Separately, 6.5 g of ginseng powder was added to 40 mL of distilled water, and then 0.5 g of yeast extract, 0.25 g of soybean peptone, and 0.5 g of calcium carbonate were added and mixed. This was used as a fermentation medium and sterilized by heating at 121°C for 15 minutes. (3) 1.5 mL of each lactic acid bacteria preculture solution was added to the fermentation medium and allowed to stand at 28°C. (4) After 6 days, 400 mg of each enzyme was added to the fermentation medium, and the mixture was shaken at 55°C and 1000 rpm to carry out the enzyme reaction for 17 hours (when only enzyme treatment was carried out, i.e., when only lactic acid bacteria fermentation treatment was carried out, this step (4) and the step (5) described below were not carried out). (5) After the enzyme reaction was completed, the mixture was heated at 90°C for 15 minutes to inactivate the enzyme. (6) After shaking well, 0.8 g of each sample was collected, to which the same volume of ethanol was added, and after stirring, the sample was centrifuged (10,000 rpm, 5 minutes) to obtain the supernatant. 1 mL of this supernatant was added to a Sep-Pak C18 cartridge (Waters) for adsorption, washed with distilled water and 40% aqueous methanol, and then eluted with methanol. (7) 10 μL of the eluted methanol solution was taken and analyzed for M1, M3, and Rb1 using high-performance liquid chromatography (HPLC) under the same conditions as in Test Example 1. The relative value to the peak area of ​​M1 when no enzyme treatment was performed and only lactic acid bacteria fermentation treatment using the A221 strain was performed, the M3 / M1 ratio (only when M1 was detected), and the ginsenoside Rb1 / M1 ratio (only when M1 was detected) were quantified.

[0096] The results are shown in the table below.

[0097] [Table 4] [Industrial Applicability]

[0098] According to the present invention, a method for producing a fermented carrot product can be provided.

Claims

1. A method for producing a fermented carrot product through an enzymatic treatment with lactase and a fermentation treatment with Lactobacillus lactic acid bacteria, The method, wherein the Lactobacillus lactic acid bacteria include Lactobacillus brevis DNBL1889 strain (Accession No. NITE AP-03235, Accession No. NITE P-03235).

2. 2. The method of claim 1, wherein the ginseng comprises at least one selected from the group consisting of Panax ginseng, Panax notoginseng, American ginseng, Bamboo ginseng, Himalayan ginseng, and Vietnamese ginseng.

3. 3. The method according to claim 1, wherein the enzyme treatment is carried out at an enzyme concentration of 0.01 to 20 w / v % at 30 to 60° C. for 3 to 72 hours.

4. The method according to any one of claims 1 to 3, wherein the Lactobacillus lactic acid bacteria further comprise at least one species selected from Lactobacillus casei and Lactobacillus brevis (excluding Lactobacillus brevis DNBL1889 strain (Accession No. NITE AP-03235, Accession No. NITE P-03235)).

5. The method according to any one of claims 1 to 4, wherein the Lactobacillus lactic acid bacteria further comprises at least one selected from the group consisting of Lactobacillus casei Hasegawa strain (FERM BP-10123), Lactobacillus casei NBRC15883 strain, and Lactobacillus brevis DNBL1871 strain (Accession No. NITE P-02661).

6. The method according to any one of claims 1 to 5, wherein the enzyme treatment is carried out simultaneously with or before the fermentation treatment.

7. The method according to any one of claims 1 to 5, wherein the fermentation treatment is carried out simultaneously with or before the enzyme treatment.

8. The method according to any one of claims 1 to 7, wherein the fermentate contains 20(S)-protopanaxadiol 20-O-β-d-glucopyranoside.

9. Lactobacillus brevis strain DNBL1889 (accession number NITE AP-03235, accession number NITE P-03235).

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