HK1118078A - Lactic acid bacteria fermented substance and fermented milk food product containing the same - Google Patents
Lactic acid bacteria fermented substance and fermented milk food product containing the same Download PDFInfo
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Description
Technical Field
The present invention relates to a lactic acid bacteria fermented product, and more particularly, to a lactic acid bacteria fermented product containing viable lactic acid bacteria at a high concentration and a fermented milk food containing the fermented product.
Background
The culture of lactic acid bacteria is carried out in various ways, and in order to produce lactic acid bacteria preparations, fermented milk, lactic acid bacteria beverages, cheese, and the like, it is most often the case that lactic acid bacteria are cultured using animal milk as a culture medium. However, since lactic acid bacteria generally have different nutritional requirements depending on their types, it is necessary to culture lactic acid bacteria for several days in order to obtain a fermented product having a sufficient acidity, even if the lactic acid bacteria are strains that do not proliferate much in a medium composed of only animal milk and have relatively excellent proliferation activity, in order to produce fermented milk, lactic acid bacteria beverages, or the like in a medium composed of only animal milk.
However, since the number of viable bacteria is reduced by culturing lactic acid bacteria for a long time, it cannot be said that the method is a preferable method for culturing lactic acid bacteria beverages, fermented milks, and the like, which are regarded as important in terms of the number of viable bacteria (the number of viable bacteria reflects various physiological effects).
In addition, in the production of various beverages and foods which are desired to have the taste of the lactic acid bacteria fermented product, since the strain to be used cannot be selected from the viewpoint of the proliferation property alone, lactic acid bacteria which are inferior in proliferation property but can provide a fermented product having a good taste may be selected and used.
Thus, it is known that, in the culture of lactic acid bacteria, various growth-promoting substances are added to a culture medium to improve the culture efficiency. Examples of the substances that are generally considered to be effective for promoting growth include: chlorella extract, iron salt, vitamins, protein decomposition product containing amino acids or peptides, yeast extract, etc.
As a technique for promoting the growth of lactic acid bacteria, the following methods have been reported in recent years: a method using a water extract of lees and/or a water extract of lees treated with a protease (patent document 1); a method using an extract extracted from leaves of a plant belonging to the genus coffea (patent document 2); a method of using a flesh portion of papaya fruits containing epidermis (patent document 3); a method of using an extract derived from algal cells of marine microalgae (patent document 4); a method using 1 or 2 or more selected from the group consisting of broccoli, cauliflower, kale, shepherd's purse, radish, nanasium, ranunculus spinosus, isatis tinctorius, cardamine grandiflora (Japanese bittersweet), wasabi, cress, mustard seed, horseradish, lily horseradish, heliotrope, coleus, turnip, rape, cabbage, spinach, pinocembrus, celery, parsley, lettuce, and apple (patent document 5); a method using 1 or more than 2 selected from the group consisting of loofah, melon, watermelon, pumpkin, sweet potato, taro, konjak, radish, carrot, tomato, green pepper, okra, shallot, cabbage, bean sprout, and orange (patent document 6); a method using an extract extracted from tea (patent documents 7 and 8); a method using a calcium salt (patent document 9); and a method using an extract of ginger, tea, or scallion (patent document 10).
On the other hand, in order to maintain the usefulness or effectiveness of lactic acid bacteria, not only the growth of bacteria is promoted, but also the death of bacteria in the lactic acid bacteria fermentation product is suppressed to improve the viability. In general, when a low-fat fermented milk food containing a lactic acid bacteria fermented product obtained by using skim milk powder or the like or lactic acid is excessively fermented, the reduction in viability of lactic acid bacteria becomes remarkable, and therefore, the problem is serious when a low-calorie fermented milk food or a fermented milk food having a low pH is produced. In order to prevent such a decrease in the viability of lactic acid bacteria and to maintain the number of lactic acid bacteria in the lactic acid bacteria fermented product, chlorella or the like is known as a material to be used.
However, in the production of beverages and foods such as lactic acid bacteria fermented products and fermented milk foods containing the fermented products, when substances added for promoting the proliferation of lactic acid bacteria or substances mixed for improving the viability of lactic acid bacteria, which have been known so far, are used to obtain a sufficient effect, the more the substances are used, the more the taste of the product is affected, and the use of the substances also leads to an increase in the cost of the product. Further, even if a state containing a large amount of live lactic acid bacteria can be maintained, the activity of lactic acid bacteria cannot be maintained in some cases, and thus a sufficient physiological effect cannot be expected.
Patent document 1: japanese patent laid-open No. 5-15366
Patent document 2: japanese laid-open patent publication No. 6-125771
Patent document 3: japanese laid-open patent publication No. 7-23777
Patent document 4: japanese laid-open patent publication No. 7-51057
Patent document 5: japanese laid-open patent publication No. 11-266860
Patent document 6: japanese laid-open patent publication No. 2-242667
Patent document 7: japanese patent No. 2667421
Patent document 8: japanese patent No. 3223326
Patent document 9: japanese patent No. 2673333
Patent document 10: japanese patent laid-open No. 2001-190272
Disclosure of Invention
The present invention has been made in view of the above circumstances, and an object of the present invention is to provide a novel substance which can easily increase the number of viable bacteria of lactic acid bacteria by merely adding and mixing the substance to a culture medium without causing a problem in taste and can maintain the number of viable bacteria in a finished product.
The present inventors have conducted intensive studies to solve the above problems, and as a result, have found that the proliferation activity of lactic acid bacteria can be easily improved without spoiling the taste of the resulting lactic acid bacteria fermented product by adding a novel extract derived from a specific plant to a medium to culture lactic acid bacteria. Further, it has been found that by culturing lactic acid bacteria in a medium containing the extract and a specific fatty acid, a lactic acid bacteria fermented product containing live lactic acid bacteria at a high concentration can be easily obtained without reducing the activity thereof. Further, the present inventors have found that various beverages and foods such as fermented milk foods produced using the lactic acid bacteria fermented products obtained by these methods do not have any influence on the taste, and have completed the present invention.
That is, the present invention is a lactic acid bacteria fermented product obtained by culturing lactic acid bacteria in a culture medium containing an extract of at least one food material selected from the group consisting of rice bran, persimmon leaf, perilla, houttuynia cordata, eucommia ulmoides, turmeric, clove, cinnamon, and sweet tea.
The present invention also provides a lactic acid bacteria fermented product obtained by culturing lactic acid bacteria in a culture medium containing an extract of at least one food material selected from the group consisting of rice bran, persimmon leaf, perilla, houttuynia cordata, eucommia ulmoides, turmeric, clove, cinnamon, and sweet tea, and oleic acid or a derivative thereof.
Further, the present invention is a fermented milk food containing the lactic acid bacteria fermented product.
Further, the present invention is a method for producing a lactic acid bacteria fermented product, characterized by culturing a lactic acid bacteria in a medium containing an extract of at least one food material selected from the group consisting of rice bran, persimmon leaf, perilla, houttuynia cordata, eucommia ulmoides, turmeric, clove, cinnamon, and sweet tea.
Further, the present invention is a method for producing a lactic acid bacteria fermented product, characterized by culturing a lactic acid bacteria in a medium containing an extract of at least one food material selected from the group consisting of rice bran, persimmon leaf, perilla, houttuynia, eucommia bark, turmeric, clove, cinnamon, and sweet tea, and oleic acid or a derivative thereof.
The extract of at least one or more food materials selected from the group consisting of rice bran, persimmon leaf, perilla, houttuynia cordata, eucommia ulmoides, turmeric, clove, cinnamon, and sweet tea used for the lactic acid bacteria fermented product of the present invention has an excellent effect of promoting the proliferation of lactic acid bacteria and improving the viability, and hardly affects the taste, so that a fermented milk food containing the lactic acid bacteria fermented product obtained by adding and mixing these substances is excellent in promoting health and highly useful as a drink and a food with less deterioration in taste.
In addition, the extract can be particularly used together with oleic acid or a derivative thereof to inhibit the death of bacteria in low-fat fermented milk food or fermented milk food with low pH, so that the viable count and the survival rate of the viable count in the product can be ensured.
Detailed Description
The lactic acid bacteria fermentation product of the present invention is obtained as follows: lactic acid bacteria were fermented under conventionally known culture conditions, except for using a medium containing an extract (hereinafter sometimes simply referred to as "extract") of at least one or more food raw materials selected from the group consisting of rice bran, persimmon leaf, perilla, houttuynia cordata, eucommia ulmoides, turmeric, clove, cinnamon, and sweet tea.
Among food materials as raw materials of the above extract, Rice bran (Rice bran) refers to a pericarp, a seed coat starch layer and a germ of a seed (brown Rice) obtained by removing a husk (Rice hull) from Rice (Oryza sativa) which is a plant belonging to the family gramineae. The rice bran is known to have the effects of enhancing immunity, preventing fatty liver, and the like.
Persimmon leaves are leaves of persimmon (Diospyros Kaki Thunb), black date (Diospyros lotus L.) or monarch emilia (Diospyros lotus L. var. glabra Makino) belonging to a plant belonging to the genus persimmon of the family of the persimmon. In the present invention, persimmon is particularly preferable among these persimmon trees. The persimmon leaf is known to have the effect of suppressing sneezing, nasal obstruction, rhinorrhea, etc.
Perilla (Perilla) is a Perilla (perillabasttescens (L.) Britton var. acuta Kudo), green Perilla (perillabastescens (L.) Britton var. acuta Kudo) or resulfur (perillabasttescens (L.) Britton var. crispo crisppa (thunnb) Decne) belonging to a plant belonging to the genus perillas of the family labiatae. In the present invention, among these plants of the genus perilla, perilla is particularly preferable. Further, leaves, branches, and seeds can be used for extraction of the extract from these perilla, but leaves are particularly preferably used. The perilla has been known to have antiallergic, hypoglycemic, youthful skin-caring effects.
Houttuynia cordata (Houttuynia cordiata Thunb.) is a plant belonging to the genus Houttuynia of the family Saururaceae. The extract obtained from houttuynia cordata may be prepared from aerial parts of the whole plant or branch, but aerial parts of the whole plant are particularly preferred. The houttuynia cordata is known to have an effect of inhibiting mucosal inflammation.
Eucommia ulmoides (Eucommia ulmoides Oliv.) is a plant belonging to the genus Eucommia of the family Eucommiaceae. Leaves or branches can be used for obtaining an extract from eucommia ulmoides, but leaves are particularly preferably used. The eucommia ulmoides is known to have the effects of regulating blood pressure, relieving stress, preventing lifestyle-related diseases, and the like.
Turmeric (curcumaric) is a rhizome of Turmeric (Curcuma longa L.) or Turmeric (Curcuma aromatica Salisb.) belonging to a plant of the genus Curcuma of the family zingiberaceae. In the present invention, among these plants of the genus curcuma, curcuma is particularly preferable. The turmeric is known to have the effects of improving liver function, preventing hangover every other day, inhibiting gastric acid secretion, and improving gastrointestinal discomfort.
Clove (cloves) is the flower bud of Clove (syzygiardia aromaticum (L.) merr. et Perry or Eugenia caryophyllata Thunb) belonging to the genus caryophylli of the family myrtaceae. The clove is known to have the effects of antisepsis, uterine contraction, toothache relief and the like.
Cinnamon (cinmamon) is the bark of Cinnamomum zeylanicum Nees or Cinnamomum cassia (cinmamum cassia Blume) belonging to Cinnamomum genus of lauraceae. Among these plants of the genus cinnamomum, cinnamomum zeylanicum is particularly preferred. Cinnamon is known to have antibacterial, cold-dispelling, antipyretic, analgesic, digestive organ activating, cold-relieving, dyspepsia relieving, diarrhea relieving, and antiemetic effects.
Sweet tea (Rubus suavissimus s.lee (Rosaceae)) is a plant belonging to the genus Rubus of the family Rosaceae. When the extract is obtained from sweet tea, leaves or stems may be used, but leaves are particularly preferably used. In recent years, the sweet tea attracts attention for anti-inflammatory and anti-allergic effects.
When the extract is obtained from the above food materials, more than 1 of the above food materials are directly or optionally subjected to washing, peeling, drying and/or pulverizing, and then extracted with solvent. These extracts may be used in a form of 1 kind or a mixture of 2 or more kinds, which is obtained by extracting each food material. Further, a mixed extract obtained by extracting a mixture obtained by mixing 2 or more kinds of food materials may be used. Of these extracts, persimmon leaf extract and sweet tea extract are preferable.
Examples of the solvent used for the extraction include lower alcohols having 1 to 5 carbon atoms such as water or ethanol, and organic solvents such as ethyl acetate, glycerin, and propylene glycol. These solvents may be used as a mixed solvent by mixing 2 or more kinds thereof. Among these solvents, water or an aqueous solvent such as water-lower alcohol is particularly preferable.
Further, the method of extracting the extract from the food material using the above-mentioned solvent is not particularly limited, but acid extraction is preferable because the acid extraction can efficiently extract the component for improving the proliferation activity of lactic acid bacteria from various food materials, and even if these extracts are added in a small amount, an excellent proliferation promoting effect can be obtained. The acid extraction is preferably carried out under acidic conditions of pH4.0 or less, wherein the acidic conditions are preferably pH3.0 to 4.0. The acid component for adjusting the pH of the solvent in the acid extraction may be any acid component, and is not particularly limited as long as it is an acid substance. Preferable examples of the acid component include organic acids such as citric acid, malic acid, tartaric acid, succinic acid, lactic acid, and acetic acid.
Further, the conditions for extracting the extract with the solvent are not particularly limited, but the extraction treatment is preferably performed at a temperature of, for example, 60 to 120 ℃, more preferably 80 to 100 ℃ for about 30 to 60 minutes.
The extract thus obtained may be used as it is, or may be used as a concentrated extract obtained by purifying and concentrating the obtained extract by ultrafiltration or centrifugation, or as a powdery extract obtained by further drying the extract by spray drying or freeze drying.
When the extract is added to a culture medium in which lactic acid bacteria can be propagated, the addition amount is preferably set after experimental confirmation, because the growth promoting effect obtained may vary depending on the strain to be cultured, the composition of the culture medium, the use of the culture, and the like, and the addition amount is preferably about 0.01 to 10% by mass (hereinafter simply referred to as "%"), more preferably about 0.01 to 5%, as an extract having a brix of about 10.
The amount of addition of these extracts may be more than 10%, but the proliferation promoting effect which is enhanced in proportion to the amount of addition may not be obtained, and the taste of the drink or food containing the culture obtained thereby may be adversely affected, and therefore, it is not preferable from the viewpoint of economy and taste. Conversely, if the amount of the extract added is less than 0.01%, the growth promoting effect cannot be sufficiently obtained, and therefore, this is not preferable.
In the present invention, oleic acid or a derivative thereof (hereinafter, simply referred to as "oleic acid") can be further added to the medium containing the extract to obtain a synergistic effect of improving the proliferation activity and viability of lactic acid bacteria. The oleic acid to be added to the medium together with the extract is not particularly limited, and examples thereof include esters in which the fatty acid moiety is oleic acid in sucrose esters, glycerin esters, sorbitan esters, propylene glycol esters, and the like, which are generally used as emulsifiers, in addition to free oleic acid or inorganic salts of oleic acid. Further, a food material containing a large amount of oleic acid may also be used. Further, if oleic acid is contained in the structure, a substance in a form such as lysolecithin may have an effect of maintaining the cell count-activity in the lactic acid bacteria fermented product of the present invention.
Specific examples of preferable oleic acids include salts of oleic acid such as sodium oleate and potassium oleate; oleic acid esters such as glyceryl oleate, polyglyceryl oleate, and sucrose oleate. Among the oleic acid esters, glyceryl oleate and polyglyceryl oleate are preferable because of their high effects of increasing the number of bacteria and improving the viability after completion of culture, and sucrose oleate is preferable from the viewpoint of physical properties such as solubility in a culture medium. These oleic acids may be used in 1 kind or in combination of 2 or more kinds.
The final concentration of oleic acid in the product is calculated by conversion to oleic acid, and preferably oleic acid is added to the medium so that the concentration is 5 to 50ppm, more preferably 5 to 25 ppm. If the oleic acid is added in an amount of less than 5ppm, the effect of the synergistic proliferation activity based on the combination with the extract and the effect of suppressing the death of bacteria in the product may sometimes be weakened. Further, if the amount is more than 50ppm, it is not preferable because it causes a problem in cost and may inhibit the growth activity of bacteria.
In the present invention, the time for adding the extract and/or the oleic acid to the medium is preferably before the fermentation of the lactic acid bacteria, but is not limited thereto, and the extract and/or the oleic acid may be added during the fermentation of the lactic acid bacteria or after the completion of the fermentation of the lactic acid bacteria. Further, the addition may be performed in several portions. It is particularly preferable to add the extract and/or oleic acid before fermentation of lactic acid bacteria, because the number of bacteria at the end of the culture and the viability of the bacteria can be maintained at a high level.
The medium to which the extract and/or oleic acid is added includes raw milk such as cow's milk, goat's milk, horse's milk, goat's milk, etc.; animal milk culture medium or various synthetic culture medium composed of skimmed milk powder, whole milk powder, milk fat, etc. The medium may further contain components generally used for a lactic acid bacterium medium. Examples of such components include: vitamins such as vitamin A, B family vitamin, vitamin C, and vitamin E; various peptides, amino acids; salts of calcium, magnesium, and the like.
In the present invention, the lactic acid bacteria used for the culture are not particularly limited as long as they are microorganisms commonly used for food production, and examples thereof include: lactobacillus casei (Lactobacillus casei), Lactobacillus acidophilus (Lactobacillus acidophilus), Lactobacillus cremoris (Lactobacillus cremoris), Lactobacillus helveticus (Lactobacillus helveticus), Lactobacillus salivarius (Lactobacillus salivatus), Lactobacillus gasseri (Lactobacillus gasseri), Lactobacillus fermentum (Lactobacillus fermentum), Lactobacillus acidopilus (Lactobacillus yoghurti), Lactobacillus delbrueckii subsp.bulgaricus (Lactobacillus delbrueckii subsp.bulgaricus), Lactobacillus delbrueckii subsp.delbrueckii), Lactobacillus johnsonii (Lactobacillus johnsonii), and the like; streptococcus bacteria such as Streptococcus thermophilus (Streptococcus thermophilus); lactococcus bacteria such as Lactococcus lactis subsp.lactis, Lactococcus lactis subsp.cremoris, Lactococcus plantarum, Lactococcus raffinosus (Lactococcus raffinosus), and Lactococcus lactis; enterococcus bacteria such as Enterococcus faecalis (Enterococcus faecalis) and Enterococcus faecium (Enterococcus faecalis). Of these lactic acid bacteria, 1 or more species of lactic acid bacteria selected from the group consisting of bacteria of the genus Lactobacillus, bacteria of the genus Streptococcus, and bacteria of the genus lactococcus are preferable. In the present invention, lactic acid bacteria are facultative anaerobic bacteria, and bacteria belonging to the genus bifidobacterium are not included as anaerobic bacteria.
In addition, the lactic acid bacteria include lactic acid bacteria that cannot sufficiently proliferate in a medium made of animal milk, and the extract used in the present invention provides a particularly significant effect in the culture of such lactic acid bacteria. Specifically, when culturing lactic acid bacteria such as Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus cremoris, Lactobacillus helveticus, Lactobacillus gasseri, Lactobacillus delbrueckii subsp.bulgaricus, Streptococcus thermophilus, and lactococcus lactis subsp.lactis, an excellent proliferation promoting effect can be obtained by adding the extract to the culture medium.
The culture conditions of the lactic acid bacteria from which the lactic acid bacteria fermented product of the present invention is obtained are not particularly limited, and for example, the lactic acid bacteria can be cultured at a temperature of about 30 to 40 ℃ for about 1 to 7 days. In addition, as the culture conditions in this case, a method suitable for culturing the lactic acid bacteria to be used can be appropriately selected from among standing, stirring, shaking, aeration, and the like.
The lactic acid bacteria fermented product thus obtained contains live lactic acid bacteria at a high concentration without reducing the activity of lactic acid bacteria. And, by mixing with other auxiliary materials which are usually allowed to be added to foods, a fermented milk food can be obtained.
Here, the fermented milk food includes fermented milk defined in the chapter of japan for dairy products and the like, beverages such as lactic acid bacteria beverages for dairy products and the like, solid yogurt (hard yogert), liquid yogurt (softyogert), plain yogurt, Kefir (Kefir), cheese and the like. The fermented milk food of the present invention also includes various drinks and foods using lactic acid bacteria, for example, fermented milks such as plain type, savory type, fruity type, sweet type, soft type, drink type, solid (hard) type, and frozen type, lactic acid bacteria drinks, kefir, cheese, and the like.
These fermented milk foods can be obtained as follows: if necessary, a sweetener such as sugar juice and other various food materials, for example, optional components such as various sugars, thickeners, emulsifiers, and various vitamins, may be added to the lactic acid bacteria fermented product. Specific examples of these food materials include: sugars such as sucrose, glucose, fructose, palatinose, trehalose, lactose, xylose, and maltose; sugar alcohols such as sorbitol, xylitol, erythritol, lactitol, palatinol (パラチニット), reduced syrup, and reduced maltose syrup; high-sweetness sweetening agents such as saccharin, sweetenin, sucralose, acesulfame potassium, stevia and the like; various thickening (stabilizing) agents such as agar, gelatin, carrageenan, pectin, xanthan gum, pectin, carob gum, gellan gum, carboxymethyl cellulose, soybean polysaccharides, propylene glycol alginate, etc.; emulsifiers such as sucrose fatty acid ester, glycerin fatty acid ester, polyglycerin fatty acid ester, sorbitan fatty acid ester, lecithin and the like; milk fat such as cream, butter, sour cream, etc.; acidulants such as citric acid, lactic acid, acetic acid, malic acid, tartaric acid, gluconic acid, and the like; various vitamins such as vitamin A, B group vitamin and vitamin C, E group vitamin; mineral components such as calcium, magnesium, zinc, iron, and manganese; flavoring agents such as yogurt, berries, oranges, pears, perilla, citrus, apples, mints, grapes, apricots, pears, custard, peaches, melons, bananas, tropical fruits, herbs, black tea, and coffee.
The fermented milk food thus obtained has a good taste and is excellent in promoting health, and is highly useful as a drink or food with little deterioration in taste. The lactic acid bacteria fermented product of the present invention has an excellent effect of promoting the growth of lactic acid bacteria and improving the viability by the extract added to the medium, and has a sufficient number of lactic acid bacteria and the number of lactic acid bacteria can be maintained. Furthermore, it was confirmed that oleic acid compounds added to the culture medium in addition to the extract had a synergistic effect on the proliferation promoting effect and the viability improving effect of lactic acid bacteria.
The action mechanism of the extract of the present invention on the promotion of the proliferation and improvement of the viability of lactic acid bacteria is not clear, but it is presumed that the extract contains a large amount of mineral components which contribute to the promotion of the proliferation and improvement of the viability of lactic acid bacteria. In the case of combining the extract and oleic acid, it is presumed that the synergistic action of the above mineral component and oleic acid promotes the proliferation of lactic acid bacteria and improves the viability.
Examples
The present invention will be described in more detail with reference to the following examples, but the present invention is not limited to these examples.
Example 1
< preparation of extract 1>
Turmeric (rhizome of Curcuma longa L.), Houttuynia cordata (Houttuynia corditta Thunb.), whole herb of the aerial part of Curcuma longa, leaves of Eucommia ulmoides (Eucommia ulmoides Oliv.), persimmon leaves (leaves of Diospyros Kaki Thunb), leaves of Perilla frutescens (L.) Britton var. acuta kudo), flower buds of clove (Syzygium aromaticum (L.) merr. Houttuynia) and cinnamon (bark of cinammom zeanicum Nees) are respectively subjected to peeling, crushing and the like, and then extracted with hot water (10 times of each raw material) at 90 ℃ for 60 minutes to prepare turmeric extract, Perilla extract, Eucommia ulmoides extract, persimmon leaf extract, clove extract and cinnamon extract. The extracts were concentrated by an evaporator, and the Brix of each extract was adjusted to 10.
Example 2
< comparison of lactic acid bacterium growth Activity >
12% of skimmed milk powder was used as a minimal medium, and 1% of the turmeric extract, houttuynia cordata extract, eucommia ulmoides extract, persimmon leaf extract, perilla extract, clove extract or cinnamon extract with Brix adjusted to 10 prepared in example 1 was added thereto, followed by sterilization, to prepare each sterile medium. A starter of 1% Lactobacillus casei YIT9029 was inoculated into each of the sterile culture media, and cultured at 37 ℃ for 48 hours. As a control medium, a product sterilized by adding 0.15% of MEAST (Saccharomyces cerevisiae autodigestate; manufactured by Japanese beer food Co., Ltd.) to a minimal medium was used. The amount of the MEAST added was set to the upper limit of the allowable range for exerting an adverse effect on the taste of the culture.
Thereafter, the proliferation activity of lactic acid bacteria was compared with the acidity of the culture (titration value in ml of 9g of the culture, from which organic acid was titrated to pH8.5 with 0.1N sodium hydroxide). The results are shown in Table 1 below.
[ Table 1]
| Minimal medium | MEAST | Turmeric extract | Houttuynia cordata extract | Eucommia bark extract | Persimmon leaf extract | Perilla extract | Clove extract | Cortex Cinnamomi extract |
| Acidity of the solution | ||||||||
| 8.2 | 10.1 | 11.1 | 10.9 | 11.0 | 11.3 | 10.7 | 10.9 | 10.7 |
Table 1 shows that the acidity of the medium supplemented with each extract of turmeric, houttuynia cordata, eucommia ulmoides, persimmon leaf, perilla, clove and cinnamon was higher than that of the medium without the extract and that of the medium supplemented with MEAST. This shows that the proliferation of lactic acid bacteria is promoted by these extracts.
Example 3
< verification of lactic acid bacterium growth Effect of acid extract >
Persimmon leaves were treated under the same conditions as in example 1 except that, instead of hot water, solutions of ph3.0, 4.0 and 5.0 prepared from water and citric acid were used to produce an extract of persimmon leaves having a brix of 10. A starter of 1% Lactobacillus casei YIT9029 was inoculated into 15% skimmed milk powder medium (containing 3% glucose) supplemented with 1% of each of the obtained extracts, and cultured at 35 ℃ for 5 days. The acidity of the resulting culture was measured in the same manner as in example 2. The results are shown in Table 2.
[ Table 2]
| Test strains | Hot water | pH3.0 | pH4.0 | pH5.0 |
| Lactobacillus casei YIT9029 | 23.1 | 24.4 | 24.5 | 23.5 |
As shown in table 2, when the extract prepared by adjusting the pH of the extraction solvent to 5.0 or less was used, the effect on the growth activity of lactic acid bacteria tended to become remarkable.
Example 4
< preparation of extract 2>
Turmeric (rhizome of Curcuma longa L.), Houttuynia cordata (Houttuynia corditta Thunb.), whole herb of the aerial part of the plant, leaves of Eucommia ulmoides (Eucommia ulmoides Oliv.), persimmon leaves (leaves of Diospyros Kaki Thunb), Perilla (perillas frutescens (L.), Britton var. acuta kudo), clove (syzyrium aromaticum (L., merr. peret.) flower bud) and cinnamon (bark of cinmamyl zeanicum Nees) were subjected to treatments such as peeling and pulverization, and then adjusted to a solution (10 times the amount of ph4.0) using water and citric acid, and extracted under the same conditions as in example 1 to prepare turmeric extract, Houttuynia cordata extract, Eucommia ulmoides extract, persimmon leaves, clove, and cinnamon extract. The extracts were concentrated by an evaporator, and the Brix of each extract was adjusted to 10.
Example 5
< verification of Lactobacillus growth Effect on extract >
The culture medium was prepared by adding 1% of turmeric extract, houttuynia cordata extract, eucommia ulmoides extract, persimmon leaf extract, perilla extract, clove extract or cinnamon extract, which had been adjusted to have a brix of 10, to 16% of skim milk powder as a minimal medium. Each medium was inoculated with 0.1% of a starter of each lactic acid bacterium and cultured at 37 ℃ for 48 hours.
In addition, Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus cremoris, Lactobacillus helveticus, Lactobacillus gasseri, Lactobacillus delbrueckii subsp bulgaricus, Streptococcus thermophilus, and Lactobacillus lactis subsp lactis were used for the culture.
Thereafter, the acidity of the culture was measured in the same manner as in example 2, and the proliferation activity of each lactic acid bacterium was compared. The results are shown in Table 3.
[ Table 3]
| Test strains | Minimal medium | Turmeric extract | Houttuynia cordata extract | Eucommia bark extract | Persimmon leaf extract | Perilla extract | Clove extract | Cortex Cinnamomi extract |
| Acidity of the solution | ||||||||
| Lactobacillus casei YIT9029 | 8.1 | 13.5 | 13.1 | 13.0 | 14.0 | 12.2 | 14.1 | 13.2 |
| Lactobacillus acidophilus YIT0070 | 9.0 | 11.4 | 10.7 | 11.1 | 11.7 | 11.4 | 10.0 | 10.4 |
| Lactobacillus cremoris YIT2002 | 1.4 | 6.7 | 6.1 | 6.4 | 7.1 | 5.8 | 6.2 | 6.1 |
| Lactobacillus helveticus YIT0100 | 17.2 | 17.8 | 17.2 | 17.5 | 17.7 | 17.4 | 17.5 | 17.6 |
| Lactobacillus gasseri YIT0192 | 2.8 | 9.1 | 9.5 | 9.8 | 10.0 | 8.1 | 8.1 | 7.2 |
| Lactobacillus delbrueckii subspecies bulgaricus YIT0098 | 14.9 | 16.1 | 15.8 | 16.4 | 16.2 | 15.4 | 16.7 | 16.1 |
| Streptococcus thermophilus YIT2001 | 7.6 | 8.9 | 8.5 | 8.9 | 8.7 | 7.9 | 8.4 | 8.1 |
| Lactococcus lactis subsp lactis YIT2013 | 6.2 | 7.2 | 6.8 | 6.9 | 7.0 | 6.2 | 6.4 | 6.5 |
Table 3 shows that, although there are some differences depending on the types of strains, the effect of these extracts on the proliferation activity of various lactic acid bacteria was confirmed in almost all strains. Particularly, significant effects are confirmed in turmeric extract, houttuynia cordata extract, eucommia ulmoides extract and persimmon leaf extract. Further, regarding the growth activity effect, it was also confirmed that a strain that grew poorly in the minimal medium tended to impart an excellent effect. This indicates that even when lactic acid bacteria that are difficult to proliferate in an animal milk medium are used, a culture with a large number of bacteria can be obtained easily by using these extracts.
Example 6
< preparation of lactic acid bacteria beverage >
A15% nonfat dry milk medium (containing 3% glucose) was used as a minimal medium, and 0.1% of each of the extracts prepared in example 4 was added thereto to prepare a test medium. These media were sterilized by heating, and then each medium was inoculated with 0.5% of a starter of Lactobacillus casei YIT9029 and cultured at 35 ℃ for 5 days to obtain a culture. The culture was homogenized at 15MPa, and 20 parts by weight of the homogenized product was added with 80 parts by weight of a 15% sugar solution sterilized at 100 ℃ for 5 minutes, and further added with 0.1% yogurt flavor (manufactured by Yakult materials Co., Ltd.) to prepare a dairy product. Taste testing of the milk products was performed by 5 experienced judges and no difference was found between each of the lactic acid bacteria beverages and the control containing the culture obtained using the minimal medium.
Further, the various extracts added to the medium were evaluated in such a way that they did not exert an influence on the taste of the minimal medium and were mixed well, and it was confirmed that the taste was not deteriorated even when they were used for culturing drinks and foods such as lactic acid bacteria beverages.
Example 7
< influence of addition amount of persimmon leaf extract on taste and proliferation Activity Effect >
(1) Preparation method of folium kaki extract
Persimmon leaf extracts were prepared under the same conditions as in example 1, respectively, using a solution (10 times the amount) of ph4.0 prepared from water and citric acid. The extracts were concentrated separately by an evaporator and adjusted to brix of 10.
(2) Confirmation of the amount added
Adding the persimmon leaf extract with the brix of 10 prepared in the step (1) into a 15% skimmed milk powder culture medium (containing 3% of glucose) in a range of 0.01-10%, and sterilizing at 100 ℃ for 60 minutes to prepare a lactic acid bacteria culture medium. This medium was inoculated with a starter of 1% Lactobacillus casei YIT9029 and cultured at 35 ℃ until the acidity (neutralization titration amount of 0.1N sodium hydroxide for 9g of sample) became 24. Thereafter, the number of lactic acid bacteria in the culture was measured using BCP medium. Then, the culture was homogenized at 15MPa, and 20 parts by weight of the homogenized product was added with 80 parts by weight of a 15% sugar solution sterilized at 100 ℃ for 5 minutes, and further added with 0.1% yogurt flavor (manufactured by yakult material corporation), to prepare a dairy product. The dairy product was subjected to taste evaluation by 5 taste panellists according to the following criteria. The results are shown in Table 4.
< evaluation criteria >
(evaluation) (Contents)
Very good: is very good
O: good taste
+ -: good without bad effect
And (delta): difference (D)
X: very poor
[ Table 4]
| Persimmon leaf extract adding amount (%) | Incubation time (hours) | Viable lactic acid count (/ ml) | Taste evaluation | |
| Without addition | 0 | 192 | 1.9×109 | ◎ |
| Water extraction | 0.01 | 147 | 2.7×109 | ◎ |
| 0.1 | 130 | 3.2×109 | ◎ | |
| 1 | 121 | 5.5×109 | ○ | |
| 5 | 116 | 5.3×109 | ○ | |
| 10 | 116 | 5.2×109 | △ | |
| Acid extraction (pH4.0) | 0.01 | 128 | 3.5×109 | ◎ |
| 0.1 | 120 | 5.4×109 | ◎ | |
| 1 | 117 | 5.7×109 | ○ | |
| 5 | 117 | 5.9×109 | ○ | |
| 10 | 115 | 5.5×109 | △ |
From table 4, it was confirmed that the addition of about 0.1% of persimmon leaf extract produced an effect of promoting the culture of lactic acid bacteria, and that the number of viable bacteria of lactic acid bacteria was also increased. On the other hand, even if 10% of the persimmon leaf extract is added to the medium, a more excellent effect proportional to the amount added cannot be obtained, but rather the taste derived from the extract tends to affect the taste of the product. In addition, compared with water extraction, the effect of acid extraction is more remarkable on the proliferation promoting effect caused by the extractum.
Example 8
< preparation of extract 3>
After rice bran (pericarp, pericarp starch layer, germ of "rice" obtained from Oryza sativa, after peeling (rice hull) is removed) (brown rice), turmeric (rhizome of Curcuma longa L.), Houttuynia cordata (Houttuynia cordita Thunb.), whole herb of overground part of eucommia ulmoides (Eucommiaulmoides Oliv.), leaves of persimmon leaves (Diospyros Kaki Thunb), perilla (perillaria Perilla frutescens (L.) Britton. var. acuta Kudo), leaves of Houttuynia cordata (syzygiardia aromaticum (L.) menn. et Perry), and cinnamon (bark of Cinnamomum zeylanicum nees) are respectively peeled, pulverized and the like, hot water (10 times of each raw material) at 80 ℃ is used to extract 60 minutes, and extracts of rice bran, perilla, and cinnamon are prepared. The extracts were concentrated separately by an evaporator and adjusted to brix of 10.
Example 9
< measurement of the number of lactic acid bacteria at the end of culture (1) >
To a minimal medium containing 15% skimmed milk powder containing 3% glucose, 1% of rice bran extract, persimmon leaf extract, perilla extract, houttuynia cordata extract, eucommia ulmoides extract, turmeric extract, clove extract or cinnamon extract having a brix adjusted to 10 prepared in example 8 was added, and then sterilized at 100 ℃ for 60 minutes to prepare each sterile medium. A1% starter of Lactobacillus casei YIT9029 was inoculated into each of the sterile culture media, cultured at 37 ℃ until pH3.7, and the viable cell count at the end of the culture was measured. Then, a medium to which sodium oleate (concentration of oleic acid is 25ppm) was added without adding the extract and a medium to which both sodium oleate and the extract were added were prepared, and the number of viable cells at the end of the culture was measured in the same manner. In addition, viable cell count was measured as follows: after the sample appropriately diluted in saline was kept in BCP medium at 37 ℃ for 3 days, the number of colonies appeared was counted. The results are shown in Table 5.
[ Table 5]
| Additive material | Number of lactic acid bacteria (cfu/ml) | Additive material | Number of lactic acid bacteria (cfu/ml) | ||
| Comparative product 1 | Without addition | 1.9×109 | Comparative product 2 | Sodium oleate | 2.1×109 |
| Example 1 | Rice bran extract | 4.7×109 | Example 2 | Rice bran extract sodium oleate | 7.1×109 |
| EXAMPLE 3 | Persimmon leaf extract | 5.2×109 | EXAMPLE 4 | Persimmon leaf extract sodium oleate | 7.8×109 |
| EXAMPLE 5 | Perilla extract | 3.6×109 | EXAMPLE 6 | Perilla extract sodium oleate | 6.6×109 |
| Example 7 | Houttuynia cordata extract | 4.2×109 | EXAMPLE 8 | Houttuynia cordata extract sodium oleate | 7.0×109 |
| Example 9 | Eucommia bark extract | 4.3×109 | Example 10 | Cortex Eucommiae extract sodium oleate | 7.4×109 |
| Example 11 | Turmeric extract | 4.2×109 | Example 12 | Turmeric extract sodium oleate | 6.8×109 |
| Example 13 | Clove extract | 4.5×109 | Example 14 | Clove extractum sodium oleate | 6.9×109 |
| Example 15 | Cortex Cinnamomi extract | 4.4×109 | Example 16 | Cortex Cinnamomi extract sodium oleate | 6.4×109 |
Table 5 shows that the synergistic increase in the number of lactic acid bacteria at the end of the culture can be obtained by combining each extract of rice bran, persimmon leaf, perilla, houttuynia cordata, eucommia ulmoides, turmeric, clove and cinnamon with sodium oleate, as compared with the case of using each extract alone or sodium oleate.
Example 10
< measurement of viable count of lactic acid bacteria in milk product (1) >
The culture produced in example 9 (comparative products 1 and 2 and examples 3 and 4) was homogenized at 15MPa, and 80 parts by weight of a 15% granulated sugar solution sterilized at 100 ℃ for 5 minutes was added to 20 parts by weight of the homogenized product, and 0.1% yogurt flavor was added to the resultant, thereby producing a dairy product. The milk product was filled into a container, and the viable cell count of the lactic acid bacteria immediately after the milk product was produced and after the milk product was stored at 10 ℃ for 14 days was measured in the same manner as in example 9. The results are shown in Table 6.
[ Table 6]
| Additive material | Number of lactic acid bacteria (cfu/ml) | ||
| Immediately after manufacture | After being stored for 14 days at 10 DEG C | ||
| Comparative product 3 | Without addition | 4.2×108 | 1.1×108 |
| Comparative product 4 | Sodium oleate 25ppm | 9.0×108 | 4.4×108 |
| Example 17 | 1 percent of persimmon leaf extract by mass | 1.0×109 | 3.8×108 |
| Example 18 | 1% by mass of persimmon leaf extract sodium oleate 25ppm | 1.8×109 | 1.1×109 |
Table 6 shows that the dairy products obtained from the cultures prepared by combining persimmon leaf extract and sodium oleate are more excellent in the effect of suppressing the variation in the number of lactic acid bacteria in the products due to storage than the dairy products obtained from the cultures not containing (without addition) or each containing these components alone.
Example 11
< measurement of viable cell count of lactic acid bacteria at the end of culture (2) >
Lactobacillus casei YIT9029 was cultured under the same conditions as in example 9 except that 1% of the persimmon leaf extract prepared in example 8 and various emulsifiers containing oleic acid to adjust the content of oleic acid to 25ppm were added to the minimal medium prepared in example 9, and the viable cell count of the resulting culture was measured by the method of example 9. The results are shown in Table 7.
[ Table 7]
| Additive material | Number of lactic acid bacteria (cfu/ml) | |
| Example 19 | Persimmon leaf extract | 2.3×109 |
| Example 20 | Persimmon leaf extract sodium oleate | 7.1×109 |
| Example 21 | Persimmon leaf extract glyceryl oleate | 7.3×109 |
| Example 22 | Persimmon leaf extract triolein pentaglycerol ester | 3.9×109 |
| Example 23 | Persimmon leaf extract hexaglycerol monooleate | 6.9×109 |
| Example 24 | Persimmon leaf extract decaoleic acid decaglycerol ester | 4.2×109 |
| Example 25 | Persimmon leaf extract sucrose oleate | 7.0×109 |
| Example 26 | Persimmon leaf extract glyceryl oleate | 3.2×109 |
Table 7 shows that the number of lactic acid bacteria in the resulting culture was increased by using oleic acid derived from any one of the emulsifiers in combination with persimmon leaf extract. Wherein, the effect is obvious when the glyceryl oleate, the hexaglycerol monooleate or the sucrose oleate is used.
Example 12
< measurement of the number of lactic acid bacteria at the end of culture (3) >
In the preparation of the extract of example 8, rice bran, persimmon leaf, eucommia bark, turmeric and clove were treated under the same conditions except that a solution prepared with water and citric acid and having pH of 3.0, 4.0 and 5.0 was used instead of hot water, to prepare extracts each having a brix of 10. Sodium oleate was added to a 15% skimmed milk powder medium to which 0.1% of the obtained extract was added so that the content of oleic acid was 25ppm, and a starter of 1% lactobacillus casei YIT9029 was inoculated and cultured at 37 ℃ until ph 3.7. The viable cell count of lactic acid bacteria in the obtained culture was measured by the method of example 9. The results are shown in Table 8.
[ Table 8]
| Additive material | pH of extraction | Number of lactic acid bacteria (cfu/ml) | |
| Example 27 | Rice bran extract sodium oleate | 5.0 | 7.1×109 |
| Example 28 | 4.0 | 9.0×109 | |
| Example 29 | 3.0 | 9.4×109 | |
| Example 30 | Persimmon leaf extract sodium oleate | 5.0 | 8.5×109 |
| Example 31 | 4.0 | 8.8×109 | |
| Example 32 | 3.0 | 9.6×109 | |
| Example 33 | Cortex Eucommiae extract sodium oleate | 5.0 | 7.0×109 |
| Example 34 | 4.0 | 8.4×109 | |
| Example 35 | 3.0 | 9.1×109 | |
| Example 36 | Turmeric extract sodium oleate | 5.0 | 7.4×109 |
| Example 37 | 4.0 | 8.5×109 | |
| Example 38 | 3.0 | 8.4×109 | |
| Example 39 | Clove extractum sodium oleate | 5.0 | 7.0×109 |
| Example 40 | 4.0 | 8.5×109 | |
| Example 41 | 3.0 | 8.4×109 |
Table 8 shows that the extract obtained by acid extraction has a tendency that the number of bacteria increases at the time of termination of culture as the pH of the solvent used for extraction is lower. In particular, the effect is more remarkable in various extracts extracted under the condition of pH5.0 or less, and more remarkable in various extracts extracted under pH4.0 or less.
Example 13
< measurement of the number of lactic acid bacteria at the end of culture (4) >
Persimmon leaf extract having a brix of 10 was prepared under the same conditions as in example 8 using a citric acid solution having a ph of 4.0. Sterilizing 10% skimmed milk powder containing 1% of the extract and 25ppm of sodium oleate to obtain sterile culture medium. The sterile medium was inoculated with 0.1% of starter of various lactic acid bacteria and cultured at 37 ℃ for 24 hours. In addition, as lactic acid bacteria, lactobacillus bulgaricus YIT0098, lactobacillus acidophilus YIT0071 and lactobacillus casei YIT9029 were used. In addition, as a control, 10% skim milk powder was used as a medium, and lactic acid bacteria were cultured in the same manner as described above. The number of lactic acid bacteria in these cultures was measured in the same manner as in example 9. The results are shown in Table 9.
[ Table 9]
| Additive material | Number of lactic acid bacteria (cfu/ml) | ||
| Additive-free culture medium | Adding culture medium | ||
| Lactobacillus bulgaricus YIT0098 | Persimmon leaf extract sodium oleate | 3.3×108 | 6.8×108 |
| Lactobacillus acidophilus YIT0071 | 3.5×106 | 2.4×107 | |
| Lactobacillus casei YIT9029 | 7.7×108 | 5.0×109 | |
Table 9 shows that the effect of increasing the number of lactic acid bacteria obtained by using a persimmon leaf extract extracted with acid together with sodium oleate is effective for each lactic acid bacterium, although the effect is somewhat different depending on the kind of lactic acid bacterium.
Example 14
< measurement of the number of lactic acid bacteria at the end of culture (5) >
Persimmon leaf extract having a brix of 10 was prepared under the same conditions as in example 8 using a citric acid solution having a ph of 4.0. This extract and oleic acid glyceride as oleic acid were added to 15% skimmed milk powder medium containing 3% glucose so as to be the addition amounts shown in the following table 10, followed by sterilization at 100 ℃ for 60 minutes to prepare respective sterile media. A starter of 1% Lactobacillus casei YIT9029 was inoculated into each of the sterile culture media, and cultured at 37 ℃ until the pH was 3.7. As a control group, 0.2% of yeast extract (manufactured by DIFCO) which is generally known as a culture promoter was added to the culture medium, and the culture was performed in the same manner. The number of lactic acid bacteria in these cultures was measured in the same manner as in example 9. The results are shown in Table 10.
[ Table 10]
Table 10 shows that the effect of increasing the number of viable bacteria was clearly confirmed by adding more than 0.1% of persimmon leaf extract and 5ppm of oleic acid at the same time, and the number was higher than that of yeast extract.
Example 15
< measurement of viable count of lactic acid bacteria in milk product (2) >
A dairy product was produced in the same manner as in example 10 using the culture produced in example 14. The dairy product was subjected to taste evaluation by 5 taste panellists according to the following criteria. The results are shown in Table 11.
< evaluation criteria >
(evaluation) (Contents)
Very good: is very good
O: good taste
+ -: good without bad effect
And (delta): difference (D)
X: very poor
[ Table 11]
Table 11 shows that when 10% of persimmon leaf extract was added to the medium, that is, 2% of persimmon leaf extract was added to the product, the taste was affected regardless of the amount of oleic acid added, indicating that the amount of oleic acid added was a tolerable limit. In addition, even with such an amount added, the taste is superior to that of the product to which the yeast extract is added.
Example 16
< preparation of extract 4>
Folium hydrangeae strigosae (Rubus suavissimus S.Lee (Rosaceae)) is processed by peeling, pulverizing, parching, and extracting with hot water (10 times of folium hydrangeae strigosae) at 90 deg.C for 60 min to obtain folium hydrangeae strigosae extract. The obtained extract was concentrated by an evaporator to adjust the Brix to 10.
Example 17
< verification of Effect on lactic acid bacteria (1) >
A sterile medium was prepared by adding 0.5% of sweet tea extract adjusted to have a brix of 10 prepared in example 16 to 12% of skimmed milk powder as a minimal medium, and sterilizing the mixture. The sterile medium was inoculated with 1% of a starter of Lactobacillus casei YIT9029 and cultured at 37 ℃ for 48 hours. As a comparative example, a medium prepared by adding 0.15% of MEAST (a self-digested product of brewery yeast, manufactured by Japanese beer food Co., Ltd.) to a minimal medium and sterilizing the resulting mixture was used. The amount of the MEAST added was set to the upper limit of the allowable range for exerting an adverse effect on the taste of the culture.
Thereafter, the proliferation activity of lactic acid bacteria was compared with the acidity of the culture (titration value in ml of 9g of the culture, from which organic acid was titrated to pH8.5 with 0.1N sodium hydroxide). The results are shown in Table 12.
[ Table 12]
| Acidity of the solution | |
| Minimal medium | 8.0 |
| MEAST | 10.0 |
| Sweet tea extract | 11.7 |
Table 12 shows that the culture medium supplemented with the sweet tea extract showed an increase in acidity compared with the culture medium supplemented with neither the additive nor the MEAST. This shows that the proliferation activity of lactic acid bacteria is promoted by the sweet tea extract.
Example 18
< verification of Effect on lactic acid bacteria (2) >
Sweet tea extracts having a brix of 10 were prepared under the same conditions except that the leaves of sweet tea were treated with an aqueous citric acid solution (temperature 90 ℃) adjusted to pH3.0, 4.0, and 5.0 instead of hot water in the preparation of the extract in example 16. A starter of 1% Lactobacillus casei YIT9029 was inoculated into 15% skimmed milk powder medium (containing 3% glucose) supplemented with 1% of each of the obtained extracts, and cultured at 35 ℃ for 5 days. The acidity of the resulting culture was measured in the same manner as in example 17. The results are shown in Table 13.
[ Table 13]
| Test strains | Hot water | pH3.0 | pH4.0 | pH5.0 |
| Lactobacillus casei YIT9029 | 23.2 | 24.6 | 24.8 | 23.8 |
As shown in table 13, when sweet tea extracts prepared by adjusting the pH of the extraction solvent to 5.0 or less were used, a tendency was observed that their effect on the proliferation activity of lactic acid bacteria became remarkable.
Example 19
< preparation of extract 5>
After the leaves of Rubus suavissimus s.lee (Rosaceae) were peeled, pulverized, and stir-fried, the extract was extracted under the same conditions as in example 16 using an aqueous citric acid solution (10 times the amount of Rubus suavissimus leaves) adjusted to ph4.0 to prepare a Rubus suavissimus extract. The obtained extract was concentrated by an evaporator to adjust the Brix to 10.
Example 20
< verification of Effect on lactic acid bacteria (3) >
A culture medium was prepared by adding 1% of the sweet tea extract having a brix adjusted to 10 of example 19 to 16% of skimmed milk powder as a minimal medium. This medium was inoculated with 0.1% of a starter of each lactic acid bacterium and cultured at 37 ℃ for 48 hours.
In addition, Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus cremoris, Lactobacillus helveticus, Lactobacillus gasseri, Lactobacillus delbrueckii subsp bulgaricus, Streptococcus thermophilus, and Lactobacillus lactis subsp lactis were used for the culture.
Thereafter, the acidity of the culture was measured in the same manner as in example 17, and the proliferation activity of each lactic acid bacterium was compared. The results are shown in Table 14.
[ Table 14]
| Test strains | Minimal medium | Sweet tea extract |
| Lactobacillus casei YIT9029 | 8.7 | 14.5 |
| Lactobacillus acidophilus YIT0070 | 9.2 | 11.4 |
| Lactobacillus cremoris YIT2002 | 1.2 | 6.5 |
| Lactobacillus helveticus YIT0100 | 17.0 | 17.0 |
| Lactobacillus gasseri YIT0192 | 2.2 | 11.0 |
| Lactobacillus delbrueckii subspecies bulgaricus YIT0098 | 14.5 | 16.5 |
| Streptococcus thermophilus YIT2001 | 7.0 | 8.2 |
| Lactococcus lactis subsp lactis YIT2013 | 6.4 | 6.8 |
Table 14 shows that although there are some differences depending on the types of strains, the effect of the sweet tea extract on the growth activity of various lactic acid bacteria was confirmed in almost all strains. Further, regarding the growth activity effect, it was also confirmed that a strain that grew poorly in the minimal medium tended to impart an excellent effect. This indicates that even when lactic acid bacteria that are difficult to proliferate in an animal milk medium are used, a fermented lactic acid bacteria product having a large number of bacteria can be easily obtained by using the sweet tea extract.
Example 21
< study on amount of sweet tea extract added >
(1) Preparation method of sweet tea extract
Sweet tea extract was prepared under the same conditions as in example 16 using an aqueous citric acid solution (10 times the amount) adjusted to ph 4.0. The resulting mixture was concentrated by an evaporator to adjust the Brix to 10.
(2) Confirmation of the amount added
Adding the sweet tea extract with the Brix of 10 prepared in the step (1) into a 15% skimmed milk powder culture medium (containing 3% of glucose) in a range of 0.01-10%, and sterilizing at 100 ℃ for 60 minutes to prepare a lactic acid bacteria culture medium. This medium was inoculated with a starter of 1% Lactobacillus casei YIT9029 and cultured at 35 ℃ until the acidity (neutralization titration amount of 0.1N sodium hydroxide for 9g of sample) became 24. Thereafter, the number of lactic acid bacteria in the culture was measured using BCP medium. Then, the culture was homogenized at 15MPa, and 20 parts by mass of the homogenized product was added with 80 parts by mass of a 15% sugar solution sterilized at 100 ℃ for 5 minutes, and further added with 0.1% yogurt flavor (manufactured by yakult material corporation), to prepare a dairy product. The dairy product was subjected to taste evaluation by 5 taste panellists according to the following criteria. The results are shown in Table 15.
< evaluation criteria >
(evaluation) (Contents)
Very good: is very good
O: good taste
+ -: good without bad effect
And (delta): difference (D)
X: very poor
[ Table 15]
| Amount of sweet tea extract (%) | Incubation time (hours) | Number of lactic acid bacteria (cfu/ml) | Taste evaluation | |
| Without addition | 0 | 184 | 1.2×109 | ◎ |
| Water extraction | 0.01 | 144 | 2.3×109 | ◎ |
| 0.1 | 123 | 3.0×109 | ◎ | |
| 1 | 120 | 4.2×109 | ◎ | |
| 5 | 118 | 4.5×109 | ○ | |
| 10 | 116 | 4.8×109 | ± | |
| Acid extraction (pH4.0) | 0.01 | 132 | 3.0×109 | ◎ |
| 0.1 | 121 | 4.2×109 | ◎ | |
| 1 | 118 | 5.1×109 | ◎ | |
| 5 | 115 | 4.9×109 | ○ | |
| 10 | 115 | 5.3×109 | ± |
From table 15, it was confirmed that the effect of promoting the culture of lactic acid bacteria by adding about 0.1% of the sweet tea extract was exhibited, and the number of viable cells of lactic acid bacteria was also increased. On the other hand, even if the sweet tea extract is added to the medium in an amount of more than 10%, the effect proportional to the amount added cannot be obtained, but the taste of the product tends to be affected. In addition, the effect of acid extraction is more remarkable than that of water extraction in terms of the proliferation promoting effect of the extract.
Example 22
< verification of Effect on lactic acid bacteria (4) >
To a minimal medium containing 15% skimmed milk powder containing 3% glucose, 1% of sweet tea extract with Brix adjusted to 10 prepared in examples 16 and 19 was added, and then sterilized at 100 ℃ for 60 minutes to prepare a sterile medium. This medium was inoculated with 1% of a starter of Lactobacillus casei YIT9029, cultured at 37 ℃ to pH3.7, and the viable cell count at the end of the culture was measured. Then, a medium to which sodium oleate (25 ppm in terms of oleic acid concentration) was added without adding the extract and a medium to which both sodium oleate and the extract were added were prepared, and the number of viable cells at the end of the culture was measured in the same manner. In addition, viable cell count was measured as follows: after the sample appropriately diluted in saline was kept in BCP medium at 37 ℃ for 3 days, the number of colonies appeared was counted. The results are shown in Table 16.
[ Table 16]
| Additive material | Number of lactic acid bacteria (cfu/ml) | |
| Comparative product 5 | Without addition | 1.7×109 |
| Example 42 | Sweet tea extract (Hot water) | 4.1×109 |
| Example 43 | Sweet tea extract (pH4.0) | 5.4×109 |
| Comparative product 6 | Sodium oleate | 2.5×109 |
| Example 44 | Sweet tea extract (hot water) sodium oleate | 5.5×109 |
| Example 45 | Sweet tea extract (pH4.0) sodium oleate | 6.5×109 |
As can be seen from table 16, the synergistic increase in the number of lactic acid bacteria was obtained by combining each sweet tea extract with sodium oleate, as compared with the case of using the sweet tea extract alone.
Example 23
< verification of Effect on lactic acid bacteria (5) >
The lactic acid bacteria fermented product (products 42, 43, 44 and 45) produced in example 22 was homogenized at 15MPa, and 80 parts by mass of a 15% sugar solution sterilized at 100 ℃ for 5 minutes was added to 20 parts by mass of the homogenized product, and 0.1% yogurt flavor was added to the resultant, thereby producing a dairy product. The milk product was filled into a container, and the viable cell count of the lactic acid bacteria immediately after the milk product was produced and after the milk product was stored at 10 ℃ for 14 days was measured in the same manner as in example 22. The results are shown in Table 17.
[ Table 17]
| Additive material | Number of lactic acid bacteria (cfu/ml) | ||
| Immediately after manufacture | After being stored for 14 days at 10 DEG C | ||
| Comparative product 7 | Without addition | 3.4×108 | 9.7×107 |
| Example 46 | 1% of sweet tea extract (hot water) | 8.2×108 | 4.4×108 |
| Example 47 | Sweet tea extract (Hot Water) 1 wt% sodium oleate 25ppm | 1.3×109 | 7.4×108 |
| Example 48 | Sweet tea extract (pH4.0) 1 wt% | 1.0×109 | 5.4×108 |
| Example 49 | Sweet tea extract (pH4.0) 1% by mass sodium oleate 25ppm | 1.4×109 | 8.4×108 |
Table 17 shows that the dairy products obtained using the lactic acid bacteria fermented product prepared by using the sweet tea extract alone or by using the sweet tea extract and sodium oleate in combination as a raw material are more excellent in the effect of suppressing the variation in the number of lactic acid bacteria in the product due to storage than the dairy products obtained using the lactic acid bacteria fermented product not containing these substances. And, unlike when the sweet tea extract is used alone, a synergistic effect of the above effects can be obtained when it is used in combination with sodium oleate.
Example 24
< verification of Effect on lactic acid bacteria (6) >
A sweet tea extract having a brix of 10 was prepared under the same conditions as in example 16 using an aqueous citric acid solution having a ph of 4.0. Adding 1% of the extract and 25ppm of sodium oleate calculated by oleic acid into 10% of skimmed milk powder, sterilizing, and making into sterile culture medium. This medium was inoculated with 0.1% of a starter of each lactic acid bacterium and cultured at 37 ℃ for 24 hours. In addition, as lactic acid bacteria, lactobacillus delbrueckii subsp bulgaricus, lactobacillus acidophilus, and lactobacillus casei were used. In addition, as a comparative example, lactic acid bacteria were cultured in the same manner as described above using 10% skim milk powder as a medium. The number of lactic acid bacteria in these cultures was measured in the same manner as in example 22. The results are shown in Table 18.
[ Table 18]
| Test strains | Number of lactic acid bacteria (cfu/ml) | |
| Additive-free culture medium | Adding culture medium | |
| Lactobacillus delbrueckii subspecies bulgaricus YIT0098 | 3.0×108 | 6.5×108 |
| Lactobacillus acidophilus YIT0071 | 3.5×108 | 6.4×108 |
| Lactobacillus casei YIT9029 | 8.2×108 | 2.4×109 |
Table 18 shows that the effect of sweet tea extract and oleic acid was slightly different depending on the kind of lactic acid bacteria, but was effective for each lactic acid bacteria.
Example 25
< verification of Effect on lactic acid bacteria (7) >
To a minimal medium of 15% skimmed milk powder containing 3% glucose, 1% of sweet tea extract adjusted to have a brix of 10 prepared in example 19 and various emulsifiers containing oleic acid such that the content of oleic acid was 25ppm were added, followed by sterilization at 100 ℃ for 60 minutes to prepare sterile media, respectively. These media were inoculated with 1% of a starter of Lactobacillus casei YIT9029, cultured at 37 ℃ to pH3.7, and the viable cell count was measured in the same manner as in example 21. The results are shown in Table 19.
[ Table 19]
| Additive material | Number of lactic acid bacteria (cfu/ml) | |
| Example 50 | Sweet tea extract | 1.8×109 |
| Example 51 | Sweet tea extract sodium oleate | 6.0×109 |
| Example 52 | Sweet tea extract oleic acid monoglyceride | 6.5×109 |
| Example 53 | Glycyrrhiza extract triolein pentaglycerol ester | 4.2×109 |
| Example 54 | Glycine max Merrill extractum hexaglycerol monooleate | 5.0×109 |
| Example 55 | Sweet tea extract tenDecaglycerol oleate | 4.2×109 |
| Example 56 | Sweet tea extract sucrose oleate | 6.4×109 |
| Example 57 | Glycerol oleate of sweet tea extract | 3.1×109 |
As shown in table 19, the effect of the proliferation activity of lactic acid bacteria was confirmed by using oleic acid derived from any emulsifier, as long as it was used in combination with the sweet tea extract. Among them, the effect is remarkable when sodium oleate, monoglyceride oleate or sucrose oleate is used.
Example 26
< verification of Effect on lactic acid bacteria (8) >
A sweet tea extract having a brix of 10 was prepared under the same conditions as in example 16 using a citric acid solution having a ph of 4.0. The extract and oleic acid glyceride as oleic acid were added to 15% skimmed milk powder medium containing 3% glucose so as to be the amounts shown in table 20 below, and then sterilized at 100 ℃ for 60 minutes to prepare respective sterile media. These sterile media were inoculated with a starter culture of 1% Lactobacillus casei YIT9029 and cultured at 37 ℃ until the pH was 3.7. As a control group, 0.2% of yeast extract (manufactured by DIFCO) which is generally known as a culture promoter was added to the culture medium, and the culture was performed in the same manner. The number of lactic acid bacteria in these cultures was measured in the same manner as in example 22. The results are shown in Table 20.
[ Table 20]
As shown in Table 20, the effect of the growth activity of lactic acid bacteria was confirmed by adding oleic acid in an amount of 0.01ppm or more.
Example 27
< verification of Effect on lactic acid bacteria (9) >
A dairy product was produced in the same manner as in example 21 using the lactic acid bacteria fermented product produced in example 26. The dairy product was subjected to taste evaluation by 5 taste panellists according to the same evaluation criteria as in example 21. The results are shown in Table 21.
[ Table 21]
Table 21 shows that, regardless of the amount of oleic acid added, when the amount of sweet tea extract added was 10%, i.e., 2% in the product, the taste was affected. In addition, even with such an amount added, the taste is superior to that of the product to which the yeast extract is added.
Industrial applicability
The lactic acid bacteria fermented product of the present invention has a large number of viable bacteria, can suppress death of lactic acid bacteria, and has little deterioration in taste. Therefore, the lactic acid bacteria fermented product can be suitably used as a raw material for various fermented milk foods.
Claims (13)
1. A fermented product of lactic acid bacteria is obtained by culturing lactic acid bacteria in a medium containing an extract of at least one food material selected from the group consisting of rice bran, persimmon leaf, perilla, houttuynia cordata, eucommia ulmoides, turmeric, clove, cinnamon, and sweet tea.
2. The lactic acid bacteria fermented product according to claim 1, wherein the extract is obtained by acid extraction.
3. The lactic acid bacteria fermented product according to claim 1, wherein the extract is obtained by acid extraction under acidic conditions of pH4.0 or less.
4. The lactic acid bacteria fermented product according to any one of claims 1 to 3, wherein the content of the extract is 0.01 to 10% by mass.
5. A lactic acid bacteria fermented product obtained by culturing lactic acid bacteria in a medium containing at least one extract of a food material selected from the group consisting of rice bran, persimmon leaf, perilla, houttuynia, eucommia bark, turmeric, clove, cinnamon and sweet tea, and oleic acid or a derivative thereof.
6. The lactic acid bacteria fermentation according to claim 5, wherein oleic acid or a derivative thereof is an oleate or a metal salt of oleic acid selected from the group consisting of glyceryl oleate, polyglycerol oleate and sucrose oleate.
7. The lactic acid bacteria fermentation according to claim 5 or 6, wherein the content of oleic acid or a derivative thereof is 1ppm to 50 ppm.
8. The lactic acid bacteria fermented product according to claim 5, wherein the extract is obtained by acid extraction.
9. The lactic acid bacteria fermented product according to claim 5, wherein the extract is obtained by acid extraction under acidic conditions of pH4.0 or less.
10. The lactic acid bacteria fermented product according to any one of claims 5 to 9, wherein the content of the extract is 0.01 to 10% by mass.
11. A fermented milk food product comprising the lactic acid bacteria fermented product according to any one of claims 1 to 10.
12. A method for preparing lactobacillus leavening is characterized in that lactobacillus is cultured in a culture medium containing at least one extract of more than one food raw material selected from rice bran, persimmon leaf, perilla, houttuynia cordata, eucommia ulmoides, turmeric, clove, cinnamon and sweet tea.
13. A method for producing a lactic acid bacteria fermented product, characterized by culturing a lactic acid bacteria in a medium containing an extract of at least one food material selected from the group consisting of rice bran, persimmon leaf, perilla, houttuynia cordata, eucommia ulmoides, turmeric, clove, cinnamon and sweet tea, and oleic acid or a derivative thereof.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2005-155583 | 2005-05-27 | ||
| JP2005-155582 | 2005-05-27 | ||
| JP2005-234747 | 2005-08-12 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| HK1118078A true HK1118078A (en) | 2009-01-30 |
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