JP7195522B2 - Method for producing ginger lactic acid bacteria fermented product - Google Patents

Description

TECHNICAL FIELD The present invention relates to a method for producing a fermented product of lactic acid bacteria ginger with improved lactic acid bacteria fermentation efficiency of ginger.

Lactic acid bacteria have been reported to have functions such as intestinal regulation, immunity enhancement, prevention of infectious diseases, and cholesterol level reduction. Actively ingesting lactic acid bacteria.

On the other hand, crops such as vegetables, fruits, and mushrooms are conventionally known to be expected to have an intestinal regulating action by containing a large amount of dietary fiber. In addition, various functionalities of these crops are being elucidated, and functional foods using these crops as raw materials are also being developed.

Recently, as a food that can be expected to have both the functionality derived from lactic acid bacteria and the functionality derived from crops such as vegetables, a food is produced by fermenting the juice or powder of vegetables with lactic acid bacteria. For example, in Patent Document 1, heat sterilized crushed vegetables are treated at a predetermined temperature with one or more lactic acid bacteria selected from the group consisting of Bifidobacterium, Lactobacillus, Pediococcus and Enterococcus. A lactic acid fermented product of crushed vegetables is described which is fermented from the start of fermentation to the end point of increasing acidity. Further, Patent Document 2 describes an acetylcholine-containing tomato lactic acid bacteria fermented food obtained by fermenting crushed tomato that has been subjected to high pressure treatment at a predetermined pressure with lactic acid bacteria. Furthermore, Patent Document 3 describes a DNA repair accelerator containing a fermented product obtained by fermenting goji berries with lactic acid bacteria belonging to the genus Lactobacillus.

On the other hand, ginger is not only used as a food flavoring, but its constituent gingerols have antipyretic, analgesic, anti-inflammatory, antioxidant, blood circulation promoting, and body temperature increasing effects. It is known to have various beneficial effects on the human body. In addition, Patent Document 4 describes that shogaols or gingerols contained in ginger promote and adjust the action of lactic acid bacteria in fermented foods and stabilize the number of lactic acid bacteria. It is shown that when ginger extract was added to the production of , the number of bacteria was significantly increased compared to the case where ginger extract was not added, and yogurt with good flavor was obtained. Furthermore, Non-Patent Document 1 describes that the addition of ginger to the fermentation system of alcohol-fermenting Zymomonas bacteria, which is a bioethanol-producing bacterium, promotes the fermentation by Zymomonas bacteria.

Thus, ginger is known to have an effective effect on the human body and an effective effect on the growth of useful bacteria. Techniques for enriching shogaols are also known in order to obtain such effective effects more efficiently. Specifically, in Patent Document 5, after mixing a raw material derived from a plant of the Zingiberaceae family with a fermented liquid, the gingerols in the raw material derived from a plant of the Zingiberaceae family are mixed with ginger for a predetermined period of time under predetermined temperature and humidity conditions. A method for enriching ginger alls for conversion to alls is described.

JP 2008-245573 A JP 2018-019629 A JP 2016-067248 A Japanese Patent Application Laid-Open No. 2005-013180 JP 2011-032248 A

Japan Society for Bioscience, Biotechnology, and Agricultural Chemistry, 2006 Chugoku-Shikoku Branch Lecture Abstracts p. 47

The present inventors focused on obtaining a fermented product of ginger using a lactic acid bacteria fermentation system. However, when lactic acid bacteria were inoculated into a base material containing ginger, lactic acid bacteria fermentation was significantly inhibited, and the production of lactic acid bacteria fermented product of ginger was seriously hindered.

Therefore, an object of the present invention is to provide a technique for improving the efficiency of lactic acid fermentation of ginger.

As a result of extensive studies, the present inventors have found that the object of the present invention is achieved by enzymatically treating a base material containing ginger with an enzyme selected from the group consisting of polyphenol oxidase, polyphenol oxygenase, and polyphenol dehydrogenase. found to be achievable. The present invention was completed by further studies based on this finding.

That is, the present invention provides inventions in the following aspects.
Section 1. Step 1 of preparing a mixture containing a fermentation base material containing ginger and/or ginger-derived components, an enzyme selected from the group consisting of polyphenol oxidase, polyphenol oxygenase, and polyphenol dehydrogenase, and lactic acid bacteria; A method for producing a ginger lactic acid bacterium fermented product, including step 2 of subjecting to fermentation conditions.
Section 2. The step 1 includes a step of adding the enzyme to the fermentation base material and performing treatment to obtain an enzyme-treated fermentation base material, and adding the lactic acid bacteria to the enzyme-treated fermentation base material to prepare the mixture. Item 2. The method for producing a fermented product of lactic acid bacteria ginger according to Item 1, comprising the step of obtaining.
Item 3. Item 2. The method for producing a fermented product of lactic acid bacteria ginger according to Item 1, wherein in the step 1, the mixture is obtained by adding the enzyme and the lactic acid bacteria to the fermentation base material at the same time.
Section 4. Item 4. The method for producing a fermented product of lactic acid bacteria ginger according to any one of Items 1 to 3, wherein the polyphenol oxidase is an enzyme having laccase activity.
Item 5. Item 4. The method for producing a ginger lactic acid bacterium fermented product according to any one of items 1 to 3, wherein the polyphenol oxygenase is an enzyme having tyrosinase activity.
Item 6. Item 4. The method for producing a ginger lactic acid bacterium fermented product according to any one of Items 1 to 3, wherein the polyphenol dehydrogenase is an enzyme having peroxidase activity.
Item 7. The lactic acid bacteria include the genus Lactobacillus, the genus Lactococcus, the genus Enterococcus, the genus Leuconostoc, the genus Streptococcus, and the genus Bifidobacterium Item 7. The method for producing a fermented product of lactic acid bacteria of ginger according to any one of items 1 to 6, which is selected from the group consisting of lactic acid bacteria belonging to the genus).
Item 8. Item 8. The method for producing a ginger lactic acid bacteria fermented product according to any one of Items 1 to 7, wherein the lactic acid bacterium is selected from the group consisting of Lactococcus lactis and Lactobacillus lactis.
Item 9. A ginger lactic acid fermented product, which is a lactic acid fermented product of a fermentation base material containing ginger and / or ginger-derived components, enzymatically treated with an enzyme selected from the group consisting of polyphenol oxidase, polyphenol oxygenase, and polyphenol dehydrogenase.
Item 10. Item 10. A food comprising the ginger lactic acid bacterium fermented product according to item 9.

ADVANTAGE OF THE INVENTION According to this invention, since the technique which improves the efficiency of lactic-acid-bacteria fermentation of ginger is provided, the manufacturing efficiency of a fermented product of lactic acid-bacteria ginger can be improved dramatically.

2 shows the results of verification of the influence of the amount of ginger used on the growth of lactic acid bacteria obtained in Test Example 2. FIG.

1. Method for producing lactic acid bacteria fermented product of ginger The method for producing lactic acid bacteria fermented product of ginger of the present invention comprises a fermentation base material containing ginger and/or ginger-derived components, and an enzyme selected from the group consisting of polyphenol oxidase, polyphenol oxygenase, and polyphenol dehydrogenase. and lactic acid bacteria; and step 2 of subjecting the mixture to lactic acid bacteria fermentation conditions. Hereinafter, the method for producing the lactic acid bacteria fermented product of ginger of the present invention will be described in detail.

Process 1
In step 1, a mixture containing a fermentation base material containing ginger and/or ginger-derived components, an enzyme selected from the group consisting of polyphenol oxidase, polyphenol oxygenase, and polyphenol dehydrogenase, and lactic acid bacteria is prepared. In the present invention, by treating the fermentation substrate with a specific enzyme in advance, lactic acid bacteria can be efficiently grown in step 2 described later, and the number of viable lactic acid bacteria can be maintained at a good level.

In an example of the aspect of step 1 (hereinafter also referred to as aspect 1a), the enzyme is added to the fermentation substrate and treated to obtain an enzyme-treated fermentation substrate; and adding the lactic acid bacteria to the fermentation base material to obtain the mixture. That is, in this aspect, the enzyme treatment is first performed on the fermentation base material, and then the lactic acid bacteria are added to the treated fermentation base material. In this aspect, the enzyme treatment and the fermentation of lactic acid bacteria by lactic acid bacteria (step 2) are distinguished in terms of process, and when the lactic acid bacteria are added, the function of the substance that inhibits or kills the lactic acid bacteria has already been deactivated. It is preferable in terms of further improving the efficiency of lactic acid fermentation of ginger.

In another example of the aspect of step 1 (hereinafter also referred to as aspect 1b), the mixture is obtained by simultaneously adding the enzyme and the lactic acid bacteria to the fermentation base material. In this aspect, since the enzymatic treatment with the enzyme and the lactic acid fermentation with the lactic acid bacterium (step 2) are allowed to proceed in parallel, adding the enzyme and the lactic acid bacterium at the same time means that the enzyme and the lactic acid bacterium are added at exactly the same timing. It is not only possible to add them at the same time, but they may be added at different timings. In the present invention, since the function of the substance that inhibits or kills the lactic acid bacteria present in the fermentation base material is deactivated by the enzyme, even when the enzyme and the lactic acid bacterium are added at the same time, the ginger can be effectively fermented with lactic acid bacteria. efficiency can be improved.

In addition, in the present invention, from the viewpoint of further improving the lactic acid bacteria fermentation efficiency of ginger, it is more preferable that the mixture is prepared in a state containing a culture medium. In this case, in Aspect 1a, the enzyme is added to the fermentation base material for treatment, and the medium can be added after the step of obtaining the enzyme-treated fermentation base material. can be added at the same time.

Fermentation Substrate Containing Ginger and/or Ginger-Derived Components In the present invention, ginger and/or ginger-derived components are used as raw materials. Ginger is a perennial plant of Zingiberaceae (Zingiber officinale), and the rhizome of ginger is used in the present invention. In addition, as for ginger, it does not matter whether it is a large variety, medium variety, or small variety. Specific ginger varieties include Sanshu Ginger, Yellow Ginger, Kintoki Ginger, Yanaka Ginger, San Omi, San Shirome, San Bushu, Sakai Beni, Nozoe No.1, Ogawa Umare, Bingo Homare, Yanagi Kogane, Tosa Taichi, Tosa no Hikari, Ogon Kokuzo, Ogon Kokuzo II and the like, preferably Tosa Taichi. In the present invention, as the ginger, a single cultivar may be used, or a plurality of cultivars may be used in combination.

In the ginger and/or ginger-derived component, the ginger rhizome itself is used as ginger, which may be in a fresh state or in a dried state. Further, the forms of ginger include crushed products, finely chopped products, ground products, and the like. When ginger itself is used as described above, it is preferable to suspend the crushed, finely chopped, or ground ginger in water or an aqueous solution, followed by cellulase treatment.

In the ginger and/or ginger-derived component, the ginger-derived component refers to a component that is a part of the constituent components of ginger and also coexists with polyphenols contained in ginger. Specific examples of ginger-derived components include ginger juice, ginger extract, and their dilutions, concentrates, and dried powders (preferably freeze-dried powders).

The method for obtaining ginger extract, its dilutions, concentrates, and dried powder (hereinafter referred to as ginger extract) is particularly limited to a method in which polyphenols are extracted together with ginger components (other than polyphenols). used without Specifically, the extraction method includes a heating extraction method, a non-heating extraction method, a supercritical extraction method, and the like.

Any of polar solvents, non-polar solvents, and mixed solvents thereof can be used as the extraction solvent. Specific examples of extraction solvents include water; alcohols such as methanol, ethanol, propanol and butanol; polyhydric alcohols such as propylene glycol and butylene glycol; ketones such as acetone and methyl ethyl ketone; chain and cyclic ethers such as tetrahydrofuran and diethyl ether; polyethers such as polyethylene glycol; hydrocarbons such as hexane, cyclohexane and petroleum ether; aromatic hydrocarbons such as benzene and toluene; fats, waxes, and other oils, preferably water, ethanol, methanol, and acetonitrile, more preferably water or ethanol, and still more preferably water. One of these solvents may be used alone, or two or more of them may be used in combination.

The extraction conditions may be appropriately determined by those skilled in the art according to the solvent used. The extraction can be carried out at 40 to 80° C. for 1 to 4 hours using 5 to 8 parts by weight of the extraction solvent. In the case of supercritical extraction, for example, 0.5 to 20 parts by weight of carbon dioxide is used with respect to 1 part by weight of ginger, and extraction is performed at 18 to 43 MPa and 20 to 40 ° C. for 0.5 to 2 hours. It can be carried out.

The fermentation substrate used in the present invention is preferably prepared as a water dilution of the ginger and/or ginger-derived components described above. The content of ginger and / or ginger-derived components contained in the fermentation base material is not particularly limited, but the dry raw material equivalent amount is 1% by weight or more, preferably 2% by weight or more, more preferably 2% by weight or more of the entire fermentation base material. 4% by weight or more, more preferably 5% by weight or more. In addition, the dry raw material equivalent amount refers to the weight of the ginger dry matter required to obtain the amount of ingredients. Therefore, when using fresh ginger, the weight of dried ginger is the equivalent amount of dried raw material, when using dried ginger, the weight of itself is the equivalent amount of dried raw material, and when using ginger extract, The weight of the dried ginger required to obtain the extract is the dry raw material equivalent amount.

In addition, the fermentation base material inevitably contains polyphenols due to the inclusion of ginger and/or ginger-derived components. The type and content of polyphenols are not particularly limited, but examples include 8 to 15% by weight of gingerol (6-gingerol) and 0.5 to 0.8% by weight of shogaol (6-shogaol). In the present invention, in order to improve the growth efficiency of lactic acid bacteria by inactivating the function of polyphenols contained in ginger by enzymatic treatment, even when using ginger and / or ginger-derived components with a high polyphenol content, It is possible to effectively improve the growth efficiency of lactic acid bacteria. From this point of view, the content of gingerol (6-gingerol) inevitably contained in the fermentation base material is 12 to 15% by weight, and the content of shogaol (6-shogaol) is 0.7 to 15% by weight. More preferably 0.8% by weight.

Fermentation base materials containing ginger and / or ginger-derived components in such a content are not inherently suitable for culturing lactic acid bacteria, and even if a lactic acid bacteria fermentation system is constructed, the number of viable lactic acid bacteria is completely reduced. Not detected or significantly reduced. In the present invention, since the function of the substance that inhibits or kills the lactic acid bacteria present in the fermentation base material is deactivated by the enzyme, the ginger and / or the ginger-derived component is such that the growth inhibition or death of the lactic acid bacteria originally occurs. Even if it is contained in a high concentration, it can effectively improve the efficiency of lactic acid fermentation of ginger.

In the present invention, when the mixture containing the fermentation base material, enzyme and lactic acid bacteria is prepared in a state containing a medium, the above-mentioned ginger and / or ginger-derived component or polyphenol is added to the fermentation base material. It is contained so that the final concentration in the state becomes the above content. Components contained in the medium include medium components such as carbon sources, nitrogen sources and inorganic salts necessary for culture and fermentation, and fermentation substrates such as glucose and lactose.

The enzymatic enzyme is selected from the group consisting of polyphenol oxidase, polyphenol oxygenase, and polyphenol dehydrogenase (hereinafter collectively referred to as polyphenol oxidase). These enzymes may be used singly or in combination. If the fermentation substrate is not treated with such an enzyme having an oxidizing activity against polyphenols, even if a lactic acid bacteria fermentation system is constructed, lactic acid bacteria will not grow at all. It causes the growth inhibition or death of lactic acid bacteria, such as the fact that they die immediately. This is believed to be due to the antibacterial activity of polyphenols such as shogaols and gingerols contained in the fermentation base material containing ginger and/or ginger-derived components. In the present invention, an enzyme having an oxidative activity against polyphenols is used to oxidatively polymerize polyphenols such as shogaol and gingerol, which are factors that inhibit the growth or kill lactic acid bacteria, and deactivate their functions (antibacterial functions). As a result, the lactic acid bacteria can be efficiently propagated in Step 2, which will be described later, and the number of viable lactic acid bacteria can be maintained at a favorable level.

The source of polyphenol oxidase is not particularly limited and includes microorganisms, plants and animals. Moreover, the polyphenol oxidase may be obtained by integrating and expressing a gene encoding the polyphenol oxidase into cells of microorganisms, plants, animals, etc. by genetic recombination.

Examples of polyphenol oxidases that can be used include commercially available enzyme preparations; part or processed products thereof; cultures of microorganisms having the activity (specifically, culture supernatants, cells, disrupted cells, etc.), or processed products thereof.

Plants, animals, basidiomycetes, parts thereof, or processed products thereof having polyphenol oxidase activity include apples (fruits), pears (fruits), avocados (fruits), bananas ( fruits), bean sprouts (hypocotyl), potato (tuber), tea (leaf), etc.; plants having polyphenol oxygenase activity include potato (tuber), etc., and basidiomycetes include mushrooms (fruiting body), etc. plants having polyphenol dehydrogenase activity include horseradish (rhizome) and the like.

The processed plant/animal/basidiomycete products and the processed culture products of microorganisms mentioned above may be subjected to any treatment that facilitates the action of polyphenol oxidase on the plants/animals/basidiomycetes or microorganisms. obtained by Such treatments include organic solvents (such as acetone), surfactants (cationic surfactants such as quaternary ammonium compounds, anionic surfactants such as sodium lauryl sulfate, and nonionic surfactants such as Tween). etc.); Enzymatic treatment with enzymes that specifically degrade cell membranes and cell walls; processing.

Furthermore, the above treated product may be used as it is, or may be used in a state of being immobilized on a carrier. Examples of the carrier include cellulose, ceramics, glass beads, etc. When using these carriers, the treated material can be immobilized by being adsorbed to the carrier. In addition to the above, examples of carriers include gelatinous substances having a lattice structure, such as agar, alginate (potassium alginate, sodium alginate, etc.), and carrageenan. An object can be immobilized by being included in a carrier.

In more specific examples of plants, animals, basidiomycetes, parts thereof, or processed products thereof having the above-mentioned activity, crushed or ground apple pulp as processed products of plants having polyphenol oxidase activity, Apple pulp juice, pear pulp crushed or ground, pear pulp juice, avocado pulp crushed or ground, banana pulp crushed or ground, bean sprout hypocotyl crushed or ground, potato tuber crushed or ground, tea leaves Examples of processed products of plants having polyphenol oxygenase activity include crushed or ground potato tubers, aqueous extracts of potato tubers, etc., and processed products of basidiomycetes include crushed mushroom fruiting bodies, Aqueous extracts of mushroom fruiting bodies and the like can be mentioned; processed products of plants having polyphenol dehydrogenase activity include crushed or ground horseradish rhizomes, aqueous extracts of horseradish rhizomes and the like.

Examples of polyphenol oxidases include enzymes with laccase activity, examples of polyphenol oxygenases include enzymes with tyrosinase activity, and examples of polyphenol dehydrogenases include enzymes with peroxidase activity. Specific enzymes having laccase, tyrosinase, and peroxidase activities are not particularly limited as long as they have laccase, tyrosinase, and peroxidase activities, respectively. Enzymes having laccase activity include laccase and enzymes other than laccase, preferably laccase. Laccase preferably includes laccase derived from basidiomycetes such as mushrooms, and particularly preferably laccase derived from fungi of the genus Trametes such as Versicolor versicolor. Enzymes having tyrosinase activity include tyrosinase and enzymes other than tyrosinase, preferably tyrosinase. The tyrosinase preferably includes tyrosinase derived from basidiomycetes such as mushrooms, and particularly preferably tyrosinase derived from mushrooms. Enzymes having peroxidase activity include peroxidase and enzymes other than peroxidase, preferably peroxidase. Examples of the peroxidase include peroxidases derived from plants of the Brassicaceae family, and particularly preferably horseradish (horseradish)-derived peroxidases.

Among these polyphenol oxidases described above, polyphenol oxidases are preferred, and among these, enzymes having laccase activity are particularly preferred.

Enzymes having laccase, tyrosinase, and peroxidase activities include commercially available enzyme preparations; parts of plants, animals, and basidiomycetes having laccase, tyrosinase, and peroxidase activities, or processed products thereof; laccase activity, tyrosinase activity, and/or Examples include cultures of microorganisms having peroxidase activity (specifically, culture supernatants, cells, disrupted cells, etc.) or processed products thereof. The method of obtaining the treated product and the mode of immobilizing the treated product on the carrier are as described above.

In step 1, the amount of polyphenol oxidase used relative to the fermentation base material containing ginger and/or ginger-derived components is not particularly limited, and can be appropriately determined according to the extent to which the lactic acid bacteria fermentation efficiency is improved. For example, the amount of polyphenol oxidase used is 2 to 1000 U, preferably 5 to 700 U, more preferably 10 to 300 U, still more preferably 20 to 100 U, more preferably 20 to 100 U, more preferably 1 mg of ginger and / or ginger-derived components in terms of dry raw materials. 20 to 50 U are preferred. Here, the unit U indicating the amount of polyphenol oxidase can be calculated from the amount of polyphenol oxidase required for the polyphenol oxidation reaction. For example, 1 U is the amount of polyphenol oxidase that can oxidize 1 μmol of substrate (polyphenol) per minute under optimum conditions (optimum temperature, optimum pH). For example, in the case of polyphenol oxidase, the unit U of the amount of polyphenol oxidase is represented as POU. 1 POU is the amount of enzyme required to increase the absorbance at 505 nm of the quinoneimine dye produced by the reaction by 0.1 in 1 minute at the beginning of the reaction. In the case of polyphenol oxygenase, tyrosinase, which is an example of polyphenol oxygenase, converts 1 μmol of tyrosine to L-DOPA per minute when the same enzyme acts on L-tyrosine at pH 6.5 and 25 ° C. 1U is the amount of enzyme that reduces the absorbance at 280 nm by 0.001 per minute when the reaction solution is 3 mL. Furthermore, in the case of polyphenol dehydrogenase, in the case of peroxidase, which is an example of polyphenol dehydrogenase, the red color of purpurogallin produced by allowing peroxidase to act on 40 mM pyrogallol and 7.4 mM hydrogen peroxide is observed by an increase in absorbance at 420 nm. The amount of enzyme that produces 1 mg of purpurogallin in 20 seconds is defined as 1 U.

In step 1, the temperature and pH when the polyphenol oxidase is brought into contact with the fermentation base material containing the above ginger and / or ginger-derived components are determined based on the optimum temperature and optimum pH of the polyphenol oxidase to be used. be done. For example, when laccase is used as the polyphenol oxidase, treatment conditions include 20-60° C. and pH 3-9. In order to obtain such pH conditions, the fermentation base containing ginger and/or ginger-derived components, or further comprising a culture medium, contains an acid such as hydrochloric acid and sodium hydroxide, potassium hydroxide, calcium carbonate, etc. The pH is adjusted accordingly using a base.

In the case of aspect 1a, in step 1, in the step of adding a polyphenol oxidase to the fermentation base material and performing treatment to obtain an enzyme-treated fermentation base material, the treatment under the above treatment conditions is performed for, for example, 2 to 24 hours. Preferably, it is carried out for 4 to 12 hours, after which lactic acid bacteria can be added to the treated fermentation substrate to obtain a mixture. Lactic acid bacteria may be added to the treated fermentation base material as it is, but the treated fermentation base material may be separated and purified by solvent extraction, solvent precipitation, or various types of chromatography. In the case of embodiment 1b, a mixture can be obtained by adding polyphenol oxidase and lactic acid bacteria to the fermentation substrate and performing the above treatment conditions for 2 to 24 hours, preferably 4 to 12 hours.

Lactic Acid Bacteria Lactic acid bacteria used in the present invention are not particularly limited, and include facultative anaerobes and anaerobes without particular limitation. Lactobacillus genus, Lactococcus genus, Enterococcus genus, Leuconostoc genus, and Streptococcus are preferred in terms of further improving the efficiency of lactic acid fermentation of ginger. genus Streptococcus and genus Bifidobacterium, more preferably Lactococcus lactis and Lactobacillus lactis. These lactic acid bacteria may be used singly or in combination.

The strain of lactic acid bacteria may be, for example, a strain distributed by a microbial strain distribution organization such as the National Institute of Technology and Evaluation (NBRC), or a fermented food containing lactic acid bacteria (eg, yogurt, etc.) fermented milk, pickles, miso, etc.) or lactic acid bacteria isolated from soil. Furthermore, in the present invention, a single isolated and purified lactic acid bacterium may be used, or the fermented food itself, which is a source of lactic acid bacteria, may be used.

A small amount of lactic acid bacteria may be included in the mixture (in this case, the lactic acid bacteria are grown in step 2), or a large amount of previously cultured lactic acid bacteria may be added. Alternatively, lactic acid bacteria resting cells collected after culturing the lactic acid bacteria in advance may be added.

Process 2
In step 2, the mixture containing the fermentation base material, polyphenol oxidase, and lactic acid bacteria obtained in step 1 is subjected to lactic acid bacteria fermentation conditions. Specifically, a lactic acid fermentation system is constructed by subjecting the mixture containing the fermentation base material to conditions suitable for lactic acid fermentation. In the mixture obtained in step 1, the function of the substance that inhibits the growth of or kills the lactic acid bacteria contained in the ginger and/or the ginger-derived component in the fermentation base material is inactivated, so a lactic acid bacteria fermentation system is constructed. As a result, a ginger lactic acid bacterium fermented product can be efficiently obtained without causing poor growth of lactic acid bacteria or a decrease in the number of viable bacteria.

In step 2, lactic acid bacteria are cultured and grown in a mixture containing a fermentation base material by lactic acid fermentation. The fermentation of lactic acid bacteria in the present invention means not only the reaction of inoculating lactic acid bacteria into foods, such as yogurt, to produce fermented foods, but also a catalyst by enzymes produced by lactic acid bacteria while growing them in a mixture containing a fermentation base material. reaction, and a cell reaction (resting cell reaction) by resting cells of lactic acid bacteria prepared in large quantities.

As the lactic acid bacteria fermentation conditions in step 2, conditions under which a lactic acid fermentation system can be established can be appropriately set according to the type of lactic acid bacteria. If the lactic acid bacteria are facultative anaerobes, the fermentation atmosphere may be air or oxygen. and other inert gases. The fermentation temperature is, for example, 20-40°C, preferably 30-38°C. In addition, the fermentation time is, for example, 24 hours to 148 hours, preferably 24 hours to 48 hours, but the end point of fermentation may be determined according to the pH and/or viable cell count of the fermented product. . Specifically, it can be the time when the pH of the fermented product is, for example, 4.5 or less, and the viable cell count is, for example, 1×10 ⁸ to 1×10 ¹⁰ .

Other processes After completion of Step 2, the fermented liquid at the end of Step 2 may be obtained as it is as a ginger lactic acid bacteria fermented product, or may be obtained as a ginger lactic acid bacteria fermented product after further passing through other steps. Other steps include concentration or powderization steps. As a method used in the concentration step, a method of heating and evaporating the fermentation liquid is common, but from the viewpoint of maintaining a larger number of viable lactic acid bacteria, it is preferable to concentrate or A method of concentration under reduced pressure and a method of concentration by membrane filtration using an ultrafiltration membrane or a hollow fiber membrane can be mentioned. Techniques used in the powderization process include adding excipients such as lactose and dextrin, followed by powderization by freeze-drying, spray-drying, or the like.

2. Lactic acid ginger fermented product The lactic acid bacterium fermented product of the present invention is a lactic acid bacterium obtained by enzymatically treating a fermentation base material containing ginger and/or ginger-derived components with an enzyme selected from the group consisting of polyphenol oxidase, polyphenol oxygenase, and polyphenol dehydrogenase. It is a fermented product. The method for producing the fermented product of lactic acid bacteria ginger of the present invention is as described in the above "1. Method for producing fermented product of lactic acid bacteria ginger".

The properties of the ginger lactic acid bacterium fermented product of the present invention include liquid, semisolid (cream, puree, powder) and solid forms.

The ginger lactic acid bacterium fermented product of the present invention is expected to possess both the functionality derived from lactic acid bacteria and the functionality derived from ginger. Therefore, the ginger lactic acid bacteria fermented product of the present invention is suitably used as a food. Foods containing the lactic acid bacteria fermented product of ginger of the present invention include food and drink such as health foods, functional foods, nutritional supplements, and foods for specified health uses. Foods containing the fermented product of lactic acid bacteria ginger of the present invention may be blended with additives that are food hygienically acceptable. Examples of such additives include sweeteners such as glucose, sucrose, fructose, isomerized liquid sugar, aspartame, and stevia; acidulants such as citric acid, malic acid, and tartaric acid; excipients such as dextrin and starch. binders, diluents such as oils and fats, perfumes, coloring agents, buffers, thickeners, gelling agents, stabilizers, preservatives, emulsifiers, dispersants, suspending agents, preservatives and the like. Specific forms of foods containing the ginger lactic acid bacteria fermented product of the present invention include supplements such as tablets, hard capsules and soft capsules, various beverages (soft drinks, carbonated drinks, beauty drinks, nutritional drinks, fruit drinks, milk drinks, etc.) and Concentrated undiluted solutions and powders for preparation of the beverages, oils and fats and processed foods, seasonings and the like can be mentioned.

EXAMPLES The present invention will be described in more detail with reference to examples below, but the present invention is not limited to these. In the following description, % means weight volume % (w/v %) unless otherwise specified.

[Culture medium]
As the MRS liquid medium (pre-culture medium), an MRS liquid medium manufactured by Becton Dickinson Co., Ltd. was dissolved in deionized water and sterilized in an autoclave at 121° C. for 15 minutes. The MRS agar medium was prepared by adding 2% agar (manufactured by Wako Pure Chemical Industries, Ltd.) to the MRS liquid medium and sterilizing it by autoclaving. As the main culture medium, 5% D-glucose, 1% soybean peptide-treated powder, and various inorganic salts were dissolved in deionized water and autoclaved at 121° C. for 15 minutes. In addition, the main culture medium was prepared at a 10-fold concentration (50% D-glucose, 10% soybean peptide-treated powder, various inorganic salts) and used as a 10-fold concentration main culture medium (x 10 main culture medium). .

[Evaluation of growth of lactic acid bacteria]
The growth of lactic acid bacteria was evaluated by measuring the number of viable bacteria by the plate pouring method and evaluating the number of bacteria by measuring the turbidity of the culture solution.

[Measurement of viable count by plate pour method]
0.05 mL of the culture broth or fermentation broth was diluted with 4.95 mL of sterilized physiological saline in 4 steps of 10-fold dilutions to obtain bacterial suspensions. The number of viable bacteria in 1 mL of these bacterial suspensions was measured. In addition, the number of viable bacteria is measured according to "Hygiene Test Method/Commentary (2005)" 1.2.1.1 Bacteria general test method 3) Bacterial count measurement (1) Pour plate culture method (p.59) went. However, MRS agar medium was used for the culture, and the culture conditions were 37° C. and 48 hours.

[Evaluation of the number of bacteria by measuring the turbidity of the culture solution]
Using a spectrophotometer UV-1800 manufactured by Shimadzu Corporation, the number of growing bacteria was evaluated by measuring the absorbance at 660 nm of the culture solution or fermentation solution diluted with physiological saline.

[Test Example 1: Verification of Effect of Improving Growth Inhibition of Lactic Acid Bacteria by Laccase Treatment]
Into 3 mL of sterilized MRS liquid medium was inoculated lactic acid bacteria of Lactococcus lactis LT57 strain that had been cryopreserved as a glycerol stock, and statically cultured at 37° C. for 1 day. 100 μL of the obtained culture solution was inoculated into 3 mL of the main culture medium and statically cultured at 30° C. for 1 day to obtain an inoculum of lactic acid bacteria.

As a fermentation base material, 250 mg of ginger powder (Nishimura Seika Co., Ltd., Tosa Tosa 1 ground product) was suspended in water to make 5 mL of ginger powder liquid (5%). Two bottles (for comparative example and example), 5 mL of water (blank: for reference) was prepared. 250 mg of a cellulase agent and 250 mg of calcium carbonate were added to each of the fermentation base materials (for comparative examples and examples) and water (blank: for reference examples), and for the ginger powder liquid for examples, 50 mg of laccase Daiwa Y120 powder (124,000 POU/g) manufactured by Daiwa Kasei Co., Ltd. was added so that 1 mg of ginger powder added was 24.8 POU. Here, regarding the unit POU representing the enzyme activity, when laccase is allowed to act on 4-aminoantipyrine and phenol at pH 4.5 and 30°C, the absorbance at 505 nm of the quinoneimine dye produced by the oxidation condensation reaction is measured for 1 minute at the initial stage of the reaction. 1 POU is the amount of laccase required to increase 0.1 to . Each was stirred at 47° C. for 4 hours at an amplitude of 120/min, and then 0.5 mL of ×10 main culture medium was added and sterilized by autoclaving. After cooling the culture medium to room temperature, 150 μL of a separately prepared inoculum of lactic acid bacteria was added, and the culture was carried out with shaking at 30° C. and an amplitude of 120/min. The viable cell count of Lactococcus lactis LT57 strain after 24 hours of culture was evaluated by measuring the viable cell count by the plate pour method.

Table 1 shows the results. As is clear from Table 1, the growth of lactic acid bacteria was confirmed in the blank containing no ginger powder (Reference Example 1), but when lactic acid bacteria were cultured in a fermentation base containing ginger powder (Comparative Example 1), The number of viable bacteria fell below the detection limit, and the growth of lactic acid bacteria was significantly inhibited. On the other hand, when the fermentation base material containing ginger powder was treated with laccase (Example 1), the same growth of lactic acid bacteria was confirmed as in the case of not containing ginger powder (Reference Example 1). In other words, when lactic acid bacteria are cultured using a medium that does not contain ginger powder as a fermentation base material, the number of viable bacteria is usually confirmed to be 1×10 ⁵ to 1×10 ⁹ cfu per 1 mL of fermentation base material. When lactic acid bacteria are cultured in a fermentation base containing ginger powder, the number of viable bacteria can be said to decrease by 1/5 to ^1/10 . It can be said that 5 to 10 ⁴ times as many viable lactic acid bacteria as without it are possible.

[Test Example 2: Verification of the effect of ginger content on the growth of lactic acid bacteria]
Ginger powder was suspended in the MRS liquid medium to final concentrations of 1%, 2.5% and 5%, and sterilized by autoclaving at 121°C for 15 minutes. After standing at 4° C. for 18 hours, the mixture was centrifuged at 5,000×g for 10 minutes to collect the supernatant. Two 3 mL aliquots of the recovered supernatant (that is, the extract (water extract) from the MRS liquid medium in which ginger powder was suspended at final concentrations of 1%, 2.5%, and 5%) were prepared, one for the test In the same manner as in Example 1, laccase Daiwa Y120 powder was added to 24.8 POU per 1 mg of ginger powder and treated with laccase, while the other was not treated with laccase. Furthermore, 5 μL of the inoculum of Lactococcus lactis LT57 strain lactic acid bacteria prepared in the same manner as in Test Example 1 was added to the sample with/without laccase treatment, and static culture was carried out at 37° C. for 48 hours. The turbidity of the grown lactic acid bacteria was measured with a spectrophotometer for absorption at 660 nm (A ₆₆₀ ).

The results are shown in FIG. As shown in FIG. 1, in the case of the water extract from the 1% suspension of ginger powder, the number of lactic acid bacteria in the case of laccase treatment (Example) was higher than that in the case of no laccase treatment (Comparative Example). The growing number increased remarkably. Such an effect was more pronounced with the water extract from the 2.5% suspension of ginger powder. Furthermore, in the case of a water extract from a suspension containing 5% ginger powder, the growth of lactic acid bacteria is significantly increased, coupled with an increase in the nutrient source derived from the ginger powder in the water extract. Admitted.

[Test Example 3: Production of ginger lactic acid bacteria fermented product using various lactic acid bacteria]
Ginger powder was added to a final concentration of 5% or 10% in the MRS liquid medium and autoclaved at 121°C for 15 minutes. After standing at 4° C. for 18 hours, centrifugation was performed at 3,000×g for 10 minutes, and the supernatant was collected. The supernatant was further centrifuged under the same conditions, and the supernatant was collected again. The recovered supernatant (that is, the extract (water extract) from the MRS liquid medium in which ginger powder was suspended at a final concentration of 5% and 10%) was dispensed by 25 mL, and the enzyme titer of laccase per 1 mg of ginger powder was measured. 250 mg of laccase Daiwa Y120 (124,000 POU/g) was added to the water extract from the 5% ginger suspension and 500 mg to the water extract from the 10% ginger suspension so that the total amount was 24.8 POU. (with laccase treatment). Furthermore, as a control, a sample without laccase was also prepared (no laccase treatment).

The lactic acid bacteria shown in Table 2 were inoculated into the MRS medium and precultured by standing at 37° C. for 48 hours. 5 μL of the preculture solution was inoculated into the laccase-treated/laccase-free aqueous extract of the above 5% or 10% ginger suspension, and statically cultured at 37° C. for 48 hours. Growth of lactic acid bacteria after culture was visually evaluated based on the following criteria.

-: Growth cannot be confirmed
+: Growth can be confirmed ++: Vigorous growth can be confirmed

Table 2 shows the results. As is clear from Table 2, when any lactic acid bacterium is used, the growth of lactic acid bacteria is significantly better in the case of laccase treatment (Example) than in the case of no laccase treatment (Comparative Example). It was confirmed that

[Test Example 4: Verification of the effect of improving inhibition of growth of lactic acid bacteria when laccase treatment and lactic acid bacteria culture are performed in parallel]
Lactococcus lactis LT57 strain was precultured in 3 mL of MRS medium by standing at 37° C. for 24 hours. Furthermore, 0.4 mL of the aqueous extract from the 10% ginger suspension described in Test Example 3 was added with 10% laccase filtered and sterilized through a 0.22 μm filter so that the enzyme titer was 31 POU per 1 mg of ginger powder. After adding 100 μL of Yamato Y120 solution (with laccase treatment) or 100 μL of sterilized water (without laccase treatment), 10 μL of the above preculture solution of Lactococcus lactis LT57 strain was inoculated, shaken at 30° C. for 48 hours, shaking speed 120. / min and fermented. After fermentation, the growth of lactic acid bacteria was evaluated according to the same criteria as in Test Example 3.

Table 3 shows the results. As is clear from Table 3, even when laccase and lactic acid bacteria were simultaneously added to the base material containing the ginger-derived component, the laccase treatment was performed as compared with the case where the laccase treatment was not performed (Comparative Example 2). In the case (Example 2), it was confirmed that the growth of lactic acid bacteria was remarkably improved.

Claims (7)

Step 1 of preparing a mixture containing a fermentation base material containing ginger and/or ginger-derived components, an enzyme having laccase activity, and lactic acid bacteria (excluding the combination of Lactobacillus plantarum and Lactobacillus brevis) and a step 2 of subjecting the mixture to lactic acid bacteria fermentation conditions. The step 1 includes a step of adding the enzyme to the fermentation base material and performing treatment to obtain an enzyme-treated fermentation base material, and adding the lactic acid bacteria to the enzyme-treated fermentation base material to prepare the mixture. The method for producing a ginger lactic acid bacterium fermented product according to claim 1, comprising the step of obtaining. 2. The method for producing a fermented product of lactic acid bacteria ginger according to claim 1, wherein in said step 1, said mixture is obtained by adding said enzyme and said lactic acid bacteria to said fermentation base material at the same time. The lactic acid bacteria include the genus Lactobacillus, the genus Lactococcus, the genus Enterococcus, the genus Leuconostoc, the genus Streptococcus, and the genus Bifidobacterium ( The method for producing a fermented product of lactic acid bacteria of ginger according to any one of claims 1 to 3 , which is selected from the group consisting of lactic acid bacteria belonging to the genus Bifidobacterium. The method for producing a fermented product of lactic acid bacteria ginger according to any one of claims 1 to 4 , wherein the lactic acid bacterium is selected from the group consisting of Lactococcus lactis and Lactobacillus lactis. Ginger lactic acid bacteria, which is a fermented product of a fermentation base material containing ginger and / or ginger-derived components, enzymatically treated with an enzyme having laccase activity (excluding a combination of Lactobacillus plantarum and Lactobacillus brevis) . fermented product. A food containing the ginger lactic acid bacterium fermented product according to claim 6 .

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