MX2007014876A - Lactic acid bacteria fermented substance and fermented milk food product containing the same. - Google Patents

Abstract

A lactic acid bacteria fermented substance is characterized by being 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 frutescens, Houttuynia cordata, Eucommia ulmoides, turmeric, clove, cinnamon and Rubus suavissimus. Because the extract to be used in the production of the lactic acid bacteria fermented substance does not cause a flavor problem and can increase the number of living lactic acid bacteria easily by simply being added to and mixed in the medium, the lactic acid bacteria fermented substance containing many living lactic acid bacteria in which high activity is maintained and a food or a drink using the same can be obtained.

Description

FERMENTED SUBSTANCE WITH THE BACTERIA OF THE LACTIC ACID AND FERMENTED LACTICAL FOOD PRODUCT THAT CONTAINS IT TECHNICAL FIELD The present invention relates to fermentation products of the lactic acid bacteria, and more specifically to fermentation products of the lactic acid bacteria, which contain viable lactic acid bacteria at high concentration, and also to fermented dairy products containing said lactic acid bacteria. Fermentation products of lactic acid bacteria.

BACKGROUND OF THE INVENTION The cultivation of lactic acid bacteria takes place in various ways; more broadly through the use of animal milk in the production of preparations of lactic acid bacteria, and also in the production of fermented milk, beverages of lactic acid bacteria, cheese, etc. However, generally, lactic acid bacteria have different auxotropes depending on the species, and in general they do not grow sufficiently in a medium consisting solely of animal milk. Therefore, even with a strain having relatively good proliferation among the lactic acid bacteria, the culture should be continued for as long as several days to obtain a fermentation product such as a fermented milk or lactic acid bacteria beverage, of sufficient acidity in its production. However, said long-term cultures of the lactic acid bacterium cause the reduction in the count of the viable cells, and therefore, it is not necessarily considered as a preferred culture method for the production of beverages of the bacterium. lactic acid, fermented milk, or the like in all of which the count of viable cells is important in the expectation of various physiological effects. On the other hand, in the production of various beverages or foods in each one the flavor of a fermentation product of the lactic acid bacteria is important, the strains to be used can not be selected only from the point of view of their proliferation. Lactic acid bacteria of low proliferation can be used in some cases for the availability of fermentation products with good taste. In the culture of lactic acid bacteria, it is, therefore, a common practice to add various growth promoting substances in advance to a medium for the purpose of improving the efficiency of the culture. Examples of growth promoting substances, which are generally considered effective, including chlorella extract, iron salts, vitamins, proteloliths including amino acids or peptides, and yeast extract. In addition, as other techniques designed to promote the growth of lactic acid bacteria, it has recently been reported MU? The method makes use of an aqueous extract of sake sediments and / or an aqueous extract of sake sediments that has been treated with a protease (Patent Document 1), a method that makes use of an extract of the leaves of a Coffea arabica plant (Patent Document 2), a method that makes use of fresh parts of papaya that include their husks (Patent Document 3), a method that makes use of an extract from algae bodies of marine microalgae (Document of Patent 4), a method that makes use of one or more vegetables or the like selected from the group consisting of broccoli, cauliflower, kale, shepherd's purse, radishes, Arabic alabra, celery buttercup leaves, white celery, mustard, Japanese bitter cress, barbarea, watercress, mustard leaf, brown mustard, wasabi (green horseradish paste), herbaceous perennial, Japanese long turnip, Japanese pickled turnip, turnip, rapeseed, cabbage, spinach, komatuna (Brassica campestric var. peruviridis), celery , parsley, lettuce and apple (Patent Document 5), a method that makes use of one or more types of vegetables or the like selected from the group consisting of scrub gourd, cucumber, sweet melon, squash, sweet potato, toran, 'KONJAK ', Japanese radish, carrot, tomato, green pepper, okra, Welsh onion, Chinese cabbage, bean sprouts and tangerine orange (Patent Document 6), a method that makes use of a tea extract (Patent Documents 7 and 8) ), a method that makes use of a calcium salt (Patent Document 9) and a method that makes use of an extract of ginger, tea or green onion (Patent Document 10), etc. ? > However, in order to maintain the usefulness or effectiveness of the lactic acid bacteria, it is necessary not only to promote the growth of the bacteria but also to reduce the death of the bacteria and improve the viability of the bacteria in the fermentation product by lactic acid bacteria. Generally, a reduction in the viability of the lactic acid bacterium becomes pronounced when low-fat fermented dairy foods containing a fermentation product of the lactic acid bacteria of skimmed milk powder or the like are prepared, or when it has been exceeded. the fermentation of lactic acid. Therefore, the reduction in viability of lactic acid bacteria becomes more serious when preparing a low-calorie fermented milk feed or a low pH fermented milk feed. Chlorella or the like is known as a usable material to avoid a reduction in viability of the lactic acid bacteria, and maintain the cell count of the lactic acid bacteria in a fermented milk feed. However, in the production of a beverage or food such as a fermentation product of the lactic acid bacterium or a fermented dairy food containing it, a conventionally known substance added to promote the growth of the lactic acid bacteria or a substance conventionally known added to improve the viability of lactic acid bacteria can affect the taste of the product itself in many cases and can also cause a rise in the cost of the product, when used in such quantity to bring enough HMá ^ il effects. Additionally, even if it is possible to maintain a state in which a large number of viable lactic acid bacteria are contained, the lactic acid bacteria can not be kept active, which makes it difficult to expect sufficient physiological effects in some cases. [Patent Document 1 JP-A. 05-015366 [Patent Document 2 JP-A-07-125771 [Patent Document 3] JP-A-07-023777 [Patent Document 4 JP-A -07-051057 [Patent Document 5 JP-A-11 -266860 [Patent Document 6 JP-A-02-242667 [Patent Document 7 JP-B-2667421 [Patent Document 8] JP-B-3223326 [Patent Document 9 JP-B-2673333 [Patent Document 10 ] JP-A-2001 -190272 BRIEF DESCRIPTION OF THE INVENTION Problems to be solved by the invention An object of the present invention is, therefore, to find a novel substance, the single addition and mixing of which to a medium makes it possible not only to easily increase the count of viable cells of the bacterium of the lactic acid but also maintain the viable cell count even after the preparation of a final product without developing problems related to taste and taste, and to use the substance for the provision of a fermentation product of a lactic acid bacteria, said fermentation product containing a number of viable lactic acid bacteria, or a beverage or food that makes use of the fermentation product.

Means for solving the problems In order to achieve the object described above, the present inventors have conducted an extensive investigation. As a result, it was found that without imparting the flavor and taste of a fermentation product to be obtained by the lactic acid bacteria, the proliferative activities of the lactic acid bacteria can be easily improved by adding a novel extract of a specific plant to a and cultivate lactic acid bacteria there. In addition, the present inventors have also found that culturing the lactic acid bacteria in a medium, which contains the aforementioned extract and a specific fatty acid, makes it possible to obtain a fermentation product of the lactic acid bacterium containing bacteria of the lactic acid bacteria. viable lactic acid at a high concentration without reducing its activities. Additionally, the present inventors have also found that various beverages or foods, such as fermented dairy foods, prepared by the methods described above are free of any problem in taste and taste, leading to complete the present invention.

In one aspect of the present invention, there is thus provided a fermentation product of the lactic acid bacterium, which has been obtained by culturing the lactic acid bacteria in a medium comprising an extract of at least one food material selected from the group consisting of rice husk, persimmon leaves, perilla, Houttuynia cordata Thunb, Eucommia ulmoides Oliv., turmeric, clove, cinnamon and Rubus suavissimus S. Lee (Rosaceae). In another aspect of the present invention, a fermentation product of the lactic acid bacterium, which has been obtained by culturing the lactic acid bacteria in a medium comprising an extract from at least one selected food material, is also provided. from the group consisting of rice husk, persimmon leaves, perilla, Houttuynia cordata Thunb, Eucommia ulmoides Oliv., turmeric, clove, cinnamon and Rubus suavissimus S. Lee (Rosaceae), and oleic acid or a derivative thereof. In a further aspect of the present invention, a fermented milk feed comprising the fermentation product described above is provided. In still another aspect of the present invention, there is also provided a method for the production of a fermentation product of the lactic acid bacterium, which comprises culturing the lactic acid bacteria in a medium comprising an extract of at least one material food selected from the group consisting of cascara iM of rice, persimmon leaves, perilla, Houttuynia cordata Thunb, Eucommia ulmoides Oliv., turmeric, clove, cinnamon and Rubus suavissimus S, Lee (Rosaceae). In yet a further aspect of the present invention there is also provided a method for the production of a fermentation product of the lactic acid bacterium, which comprises culturing the lactic acid bacteria in a medium comprising an extract of at least one material food selected from the group consisting of rice husk, persimmon leaves, perilla, Houttuynia cordata Thunb, Eucommia ulmoides Oliv., turmeric, clove, cinnamon and Rubus suavissimus, Lee (Rosaceae), and oleic acid or a derivative thereof.

Effect of the Invention The extract, which is useful in the fermentation product of the lactic acid bacteria of the present invention and has been derived from at least one food material selected from the group consisting of rice husk, persimmon leaves, knob, Houttuynia cordata Thunb, Eucommia ulmoides Oliv., turmeric, clove, cinnamon and Rubus suavissimus S ^ Lee (Rosaceae), has excellent growth promoting effects or viability enhancing effects for lactic acid bacteria and, moreover, has practically no effect on taste and taste. A fermented milk food, which has been obtained by adding and mixing the extract and containing the fermentation product of the lactic acid bacteria, is therefore excellent for the promotion of health, and has a great utility as a beverage or food that does not undergo a great deterioration in taste and taste. In particular, the combined use of the aforementioned extract with oleic acid or a derivative thereof can reduce the death of the bacterium even in a low-fat fermented milk feed or in a low-fermented milk feed, thereby guaranteeing the count of viable cells in the product and its viability.

DETAILED DESCRIPTION OF THE INVENTION The fermentation product of the lactic acid bacterium of the present invention is obtained by fermenting the lactic acid bacteria under conventionally known culture conditions, except by the use of a medium containing an extract of at least one food material selected from the group consisting of rice husk, persimmon leaves, goatee, Houttuynia cordata Thunb, Eucommia ulmoides Oliv., turmeric, clove, cinnamon and Rubus suavissimus, Lee (Rosaceae) (in the following, it will simply be referred to as "an extract") . Among the food materials that can each be used as a raw material for the aforementioned extract, the rice husk is a mixture of perocarp, aleurone layers and grain germs (brown rice) available from Oryza satvia without the outer layer of the rice without and leaves and branches can be used, the use of leaves being particularly preferred. The Eucomia ulmoides Oliv. It is known to have effects such as blood pressure control, stress relief and prevention of lifestyle related diseases. Turmeric is the rhizome of Curcuma lon a L. or Turmeric aromatic Salisb. In the present invention, Curcuma longa L. is particularly preferred among plants belonging to Turmeric. Curcuma longa L. is known to have effects such as hepatic function improving effects, hangover preventive effects, gastric antisecretory effects and gastrointestinal dysfunction improving effects. The clove is the bud of Syzvgium aromaticum (L.) Merr et Perry or Eugenia caryophyllata Thunb. The nail is known to have conservative effects, uterine contraction activities, dental pain reducing effects, and the like. Cinnamon is the bark of Cinnamomum zeylanicum Nees or Cinnamomum cassia Blume. Cinnamomum zeylanicum Nees is particularly preferred among these types of cinnamon plants. Cinnamon is known to have effects such as antibacterial activities, warming effects of the body, antipyretic effects, effects of activation of the digestive system, effects of improvement of various cold symptoms, release of indigestion, release of diarrhea and release of nausea.

The Rubus sauvissimus S. Lee (Rosaceae) is a plant belonging to Rubus. To obtain an extract of Rubus sauvissimus S. Lee (Rosaceae), the leaves and stem can be used, the use of the leaves being particularly preferred. Rubus sauvissimus S. Lee (Rosaceae) has attracted attention in recent years for its anti-inflammatory activities and antiallergic effects. To obtain an extract of one or more of the food materials described above, it is only necessary to extract with a solvent the food material or materials either as they are or after optionally applying processes such as washing, debarking, drying and / or grinding. Said extracts can be used either alone or in combination. A mixed extract can also be used, which is obtained by mixing a plurality of food materials and extracting them. Among these extracts, extracts of persimmon leaves and an extract of Rubus sauvissimus S. Lee (Rosaceae) are preferred. Solvents usable in extraction include water and organic solvents such as lower alcohols having from 1 to 5 carbon atoms, example, ethanol, ethyl acetate, glycerol and propylene glycol. Two or more of these solvents can be used together as a mixture of solvents. Among these solvents, water and aqueous solvents such as water-lower alcohols are particularly preferred. ÉÉÉÜ No particular limitation is imposed on the method of extracting an extract from one or more of the aforementioned food materials with the aforementioned solvent, but acid extraction is preferred since it can be efficiently extracted from the food material or materials components that can improve the proliferative activities of lactic acid bacteria and can also provide excellent growth promoting effects even when the extract is added in a small amount. The acid extraction can preferably be carried out under an acidic condition of pH 4.0 or lower, especially pH 3.0 to 4.0. No particular limitation is imposed on the acidic ingredient adapted to regulate the pH of the solvent in this acid extraction, and any ingredient can be used as long as it is acidic. Among said acidic ingredients, organic acids such as citric acid, malic acid, tartaric acid, succinic acid, lactic acid and acetic acid are preferred. Additionally, the extraction conditions for the extract with the use of the aforementioned solvent are not particularly limited, and the extraction process can be carried out, for example, by treatment for 30 to 60 minutes preferably at 60 ° C or 120 ° C , more preferably at 80 ° C to 100 ° C. The extract obtained as described above can be used as a solution as obtained immediately after extraction, or as a concentrated extract obtained by purification and concentration of the extract obtained by means of ultrafiltration, centrifugation or the like, or as and a powder extract obtained by a subsequent drying of the concentrated extract by means of spray drying, freeze drying or the like. By adding the aforementioned extract to a medium where the lactic acid bacterium can grow, its amount can preferably be determined after an experimental verification because the resulting growth promoting effects may differ depending on the strains to be cultivated, the composition of the medium and the application of the cultivated product. However, in general, the extract may be added in an amount of preferably from about 0.01 to 10% by weight (in the following, simply referred to as "%"), more preferably from about 0.01% to 5% calculated in terms of an extract which has 10 degrees Brix (sugar content). These extracts may be added in an amount greater than 10% or more. However, the growth promoting effects may not be as much proportional to the amount added. On the contrary, said excessively large amount of the extract may affect the taste and taste of the food or food that the resulting medium contains. Therefore, it is not preferred to add the extract in said excessively large amount. On the other hand, an amount of said extract less than 0.01%, may not provide sufficient growth promoting effects and, therefore, is not preferred.

In the present invention, it is possible to obtain effects without energetic growth promoters and viability enhancing effects for the lactic acid bacterium by adding oleic acid or a derivative thereof (in the following, simply referred to as "oleic acid") medium that contains the extract. No particular limitation is imposed on the oleic acid to be added together with the extract to the medium, and as an illustration are free oleic acid, inorganic salts of oleic acid, sugar esters, glycerides, sorbitan esters and propylene glycol esters, which are They commonly use as emulsifiers, and they contain oleic acid as their fatty acid portion. It is also possible to use food materials that contain a large amount of oleic acid. However, it should be noted that among those that contain oleic acid in their structures, those that have such a form as lysolecithin or the like may not be able to obtain the effects or maintain the bacterial cell count and activities in the fermentation product of the lactic acid bacteria of the invention. Specific preferred examples of oleic acid include oleate salts such as sodium oleate and potassium oleate, and oleate esters such as glyceryl oleate, polyglyceryl oleate ester and sucrose oleate.

Among the oleate esters described above, glyceryl oleate or polyglyceryl oleate is preferred because of its high effects in increasing the cell count and improving viability at the end of the culture. From the point of view of physical properties such as solubility in the medium, sucrose oleate is preferred. These oleic acids can be used alone or in combination.

The oleic acid can preferably be added to a medium in such an amount that its final concentration in the product becomes from 5 to 50 ppm, preferably 5 to 25 ppm in terms of oleic acid. An amount of oleic acid less than 5 ppm may not be able to sufficiently exhibit the non-energetic effects of growth activation and suppression of bacterial death in the product when used in combination with the extract. On the other hand, an amount of oleic acid greater than 50 ppm, can develop a problem in cost and can inhibit the proliferation of the bacteria, and therefore, is not preferred. In the present invention, the time to add the extract and the oleic acid to a medium can preferably be, but is not limited to, prior to fermentation by the lactic acid bacteria. They can also be added during fermentation by the lactic acid bacteria, or after finishing the fermentation by the lactic acid bacteria. They can be added in different portions. It is particularly preferred to add the extract and the oleic acids before fermentation by the lactic acid bacteria, because the cell count and the viability of the bacterium upon termination of the culture can be maintained at high levels. The medium to which the extract and the oleic acid are added include animal milk medium composed of fresh milk such as cow's milk, goat's milk, horse's milk and sheep's milk or dairy products such as skimmed milk powder, powder whole milk and fresh cream, and various synthetic media. These means can be those that contain kkA? l ingredients that are used in ordinary media for lactic acid bacteria. Such ingredients include, for example, vitamins such as Vitamin A, vitamins B, vitamin C and vitamin E, various peptides and amino acids, and salts such as calcium salts and magnesium salts. In the present invention, no particular limitation is imposed on the lactic acid bacteria to be used for the culture as long as it is a microorganism commonly used in food production. Illustrations of bacteria of the Lactobacillus genera such as Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus cremoris. Lactobacillus halveticus, Lactobacillus salivarius. Lactobacillus gasseri, Lactobacillus fermentum, Lactobacillus yoghurti, Lactobacillus delbrueckii subsp. bulgaricus Lactobacillus delbrueckii subsp. delbrueckii and Lactobacillus ¡ohnsonii. bacteria of the genus Streptococcus such as Streptococcus thermophilus. bacterium of the genus Lactococcus such as Lactococcus lactis subsp. lactis, Lactococcus lactis subsp. cremoris, Lactococcus plantarum and Lactococcus raffinolactis, bacteria of the genus Enterococcus such as Enterococcus faecalis and Enterococcus faecium. Among these lactic acid bacteria, it is preferred to use one or more species selected from the group consisting of the bacterium of the genus Lactobacillus, the bacterium of the genus Streptococcus and the bacterium of the genus Lactococcus. It should be noted that the term "lactic acid bacteria" as used herein means facultative anaerobic bacteria, and does not include bacteria of the genus Bifidobacteria, which are anaerobic bacteria.

The aforementioned lactic acid bacterium also includes those that do not grow sufficiently with the medium composed of animal milks, and the extract for use in the present invention particularly brings about remarkable effects in the culture of said bacteria. Specifically, excellent growth promoting effects can be obtained when the extract is added to the medium with the culture of 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 No particular limitations are imposed on the culture conditions for the lactic acid bacteria to obtain the lactic acid fermentation product of the present invention. However, for example, the culture can be conducted at about 30 to 40 ° C for 1 to 7 days. As additional conditions for said culture, a suitable method for cultivating the lactic acid bacteria can be selected from a stationary method, by agitation, by shaking, by aeration or the like. The fermentation product lactic acid bacteria obtained as described above contains viable lactic acid bacteria at a high concentration without a reduction in their proliferation. This product can be mixed with other auxiliary materials, the addition of which to food is generally approved, to produce fermented dairy foods. tib i The term "fermented dairy foods" includes fermented milks, dairy products, beverages such as lactic acid bacteria drinks, hard yogurt, soft yogurt, pure yogurt and in addition, kefir, cheese, etc., are defined by the Ministerial Ordinance in relation to the Composition Regulations, etc., for Milk and Dairy Products. The fermented dairy foods of the present invention, therefore, include various beverages and foods that make use of various lactic acid bacteria, for example, fermented milks, beverages of the lactic acid bacteria, kefir, cheese and the like, which can to be of the pure type, flavored type, fruity type, sweetened type, soft type, type of beverage, solid type (hard) or frozen type. These fermented dairy foods are obtained by adding, to the fermentation product of the lactic acid bacteria described above, a sweetener such as starch syrup and various other food materials, for example, optional ingredients such as various carbohydrates, thickeners, emulsifiers and various vitamins, as required. Specific examples of these food materials include carbohydrates such as sucrose, glucose, fructose, paratinose, trehalose, lactose, xylose and maltose; glycoalcohols such as sorbitol, xylitol, erythritol, lactitol, palatinate, reduced starch syrup and reduced maltose syrup; high intensity sweetening sweeteners such as aspartame, thaumatin, sucralose, acesulfame K and stevia; various thickeners (stabilizers) such as agar, gelatin, carrageenan, guar gum, xanthan gum, pectin, gum iüiUá i! of locust bean, gellan gum, carboxymethylcellulose, soybean polyaccharides and propylene glycol alginate; emulsifiers such as sucrose fatty acid esters, glycerin fatty acid esters, polyglycerin fatty acid esters, sorbitan fatty acid esters and lecithin; milk fats such as cream; butter and cream acid; acidic seasonings such as citric acid, lactic acid, acetic acid, malic acid, tartaric acid, and gluconic acid; various vitamins such as vitamin A, vitamins B, vitamin C and vitamin E; minerals such as calcium, magnesium, zinc, iron and manganese; and flavorings such as yogurt, berries, orange, Chinese quince, goat, citrus, apple, mint, grape, apricot, pear, cream, peach, melon, banana, tropical, herbs, black tea and coffee. The fermented milk food obtained as described above has a high utility as a beverage or food, which has good flavor and taste, is excellent in health promotion, and does not suffer much deterioration in flavor and taste. Additionally, the lactic acid fermentation product of the present invention has excellent growth promoting effects and viability enhancing effects for lactic acid bacteria due to the extract added to the culture, and therefore has and maintains a sufficient count of the bacteria of lactic acid. When the oleic acid or the like is incorporated into the medium in addition to the extract, synergistic effects are recognized with respect to the growth promoting effects and viability enhancing effects of the lactic acid bacteria.

Although the mechanism of action of the extract in the growth promoting effects and viability enhancing effects of the lactic acid bacteria in the present invention have not yet been elucidated, it is presumed that the extract contains abundant minerals, and these materials contribute to the promotion of growth and improve the viability of lactic acid bacteria. It is also presumed that, when the extract is combined with oleic or similar acids, the effects of the minerals and oleic acid or the like reach the promotion of growth and improve the viability of the lactic acid bacteria.

EXAMPLES The present invention will be described in the following in greater detail based on Examples. However, it should be kept in mind that the present invention is not limited in any way to the following examples.

EXAMPLE 1 Preparation of Extract 1 Turmeric (the curcuma of Curcuma longa L), the aerial part of the grass of Houttunynia cordata Thunb .. leaves of Eucommia ulmoides Oliv .. leaves of persimmon (leaves of Diospyros Kaki Thunb leaves of Perilla frutescens (L.) Britton var. acuta Kudo, clove (the cocoon of Svzgium aramaticum (L.) Merr. and Perry) and cinnamon (the bark of Cinnamomum zeylanium Nees) were each separately subjected to processing such as debarking and crushing, and then extracted for 60 minutes with water hot to 90 ° C (in an amount 10 times the weight of the corresponding raw material) to prepare extracts of turmeric, Houttunynia cordata Thunb., Eucommia ulmoides Oliv .. leaves of persimmon, knob, clove and cinnamon, respectively. one concentrated separately at 10 degrees Brix in an evaporator.

EXAMPLE 2 Comparison in the Proliferation of Lactic Acid Bacteria As a basal medium, 12% skim milk powder was supplied. The extracts of turmeric, Houttunynia cordata Thunb. Eucommia ulmoides Olív. Persimmon leaves, knob, clove and cinnamon, which were prepared and adjusted to 10 degrees Brix in Example 1, were added to 1% aliquots of the basal medium, respectively, followed by sterilization to prepare a sterilized medium. To each of these media, an initiator of Lactobacillus casei YIT9029 at 1% was inoculated, and the bacterial strain was then cultured at 37 ° C for 48 hours. As a comparative means, one prepared was used when adding "MEAST" (registered trademark for autolized beer yeast); product of Asahi Food and Healthcare Co., Ltd.) at 0.15% to the basal medium and then sterilizing the medium. The added amount of "MEAST" is the upper limit of a scale at which its adverse effects on the taste and taste of the crop are acceptable. The proliferations of the lactic acid bacteria in the respective cultures were then compared based on the acidity of the cultures (titration values of caustic soda to OJ N when portions (9 g) of the respective cultures and portions of an acid were taken. organic in the respective cultivated portions of caustic soda to OJ N until a pH of 8.5 was reached: unit: mL) as indices. The results are shown below in Table 1.

TABLE 1 Medium MEAST Extract Extract Extract Extract Extract Extract I Basal of leaf of Clove of Turmeric Houttuvnia Eucommia Persimmon Knob Cinnamon 'cordata Ulmoides I Thunb. Oliv Acidity i 8. 2 10.1 1 1 1 10.9 11 .0 1 1 .3 10.7 10.9 10.7 As is clear from Table 1, it has been confirmed that the acidity that is becomes greater in a medium with an extract of turmeric, Houttunynia cordata Thunb., Eucommia ulmoides Oliv .. leaves of persimmon, knob, clove or cinnamon, added to the same, that in a medium without the addition of any extract or a medium with "MEAST" added to it. This indicates that the growth of lactic acid bacteria can be promoted by these extracts.

EXAMPLE 3 Verification of the Effects of Acid Extraction Extract on the Proliferation of Lactic Acid Bacteria Under similar conditions as in the preparation of the extract in Example 1 except for the use of water and aqueous solutions, the pH of which have been adjusted to 3.0, 4.0 and 5.0, respectively, with citric acid instead of hot water, They treated the persimmon leaves. To aliquots of a 15% nonfat milk powder medium (with 3% glucose contained therein), said aliquot containing the extracts thus obtained added thereto at 1%, respectively, the Lactobacillus casei initiator YIT9029 was inoculated at 1%. The strain of bacteria was then cultured at 35 ° C for 5 days. The acidity of the resulting cultures was measured in a similar manner as in Example 2. The results are shown in Table 2.

TABLE 2 As shown in Table 2, it has been confirmed that the proliferation of lactic acid bacteria tends to become noticeable with an extract obtained by adjusting the pH of an extraction solvent to 5.0 or less.

EXAMPLE 4 Preparation of Extract 2 Turmeric (curcuma rhizome longa L), aerial part of Houttunynia cordata Thunb. Herb, leaves of Eucommia ulmoides Oliv .. persimmon leaves (leaves of Diospyros Kaki Thunb ?, leaves of Perilla frutescens (LJ Britton var. acuta Kudo, clove (the cocoon of Svzgium aramaticum (LJ Merr. and Perry) and cinnamon (the bark of Cinnamomum zeylanium Nees), were each separately subjected to processing such as debarking and crushing, and then extracted under conditions similar to Example 1 except for the use of water and an aqueous solution, the pH of which was adjusted to a pH of 4.0 with citric acid, (in quantities of at most 10 times the weight of the corresponding crude material) to prepare extracts of turmeric Houttunynia cordata Thunb. Eucommia ulmoides Oliv., Leaves of persimmon, knob, clove and cinnamon, respectively, were each separately concentrated at 10 degrees Brix, in an evaporator.

EXAMPLE 5 Verification of the Effects of the Extract on the Proliferation of Lactic Acid Bacteria As a basal medium, 16% skim milk powder was supplied. The turmeric extract, Houttunynia cordata Thunb., Eucommia ulmoides Oliv., Persimmon leaves, knob, clove and 1% cinnamon, which was adjusted to 10 degrees Brix in Example 4, were added to 1% aliquots of the basal medium. to prepare the medium, respectively. Within each of these media, primers of various strains of lactic acid bacteria were inoculated to 0.1%, and those strains of bacteria were cultured at 37 ° C for 48 hours. In the above culture, Lactobacillus casei, Lactobacillus acidophilus were used. Lactobacillus cremoris, Lactobacillus helveticus, Lactobacillus gasseri. Lactobacillus delbrueckii subsp. bulgaricus Streptococcus thermophilus and Lactococcus lactis subsp. lactis The acidity of the resulting cultures was measured in a manner similar to that of Example 2 to compare the proliferation of the various lactic acid bacteria. The results are shown in Table 3.

TABLE 3 As is clear from Table 3, the effects of these extracts on the proliferation of various lactic acid bacteria were confirmed with substantially all strains, although these vary depending on the species of the strains. Notable effects were confirmed particularly with the extracts of turmeric, Houttunynia cordata Thunb., Eucommia ulmoides Oliv., And persimmon leaves. In addition, it was confirmed that the proliferative effects have a tendency to give excellent effects to strains that are poor in the proliferation in the basal medium. This suggests that although it is difficult for lactic acid bacteria to grow in an animal environment, the use of these extracts may make it possible to easily obtain a culture of a large cell count.

EXAMPLE 6 Preparation of Lactic Acid Bacteria Drink A 15% skimmed milk powder medium (with 3% glucose contained therein) was supplied as a basal medium. The various extracts prepared in Example 4 were added 0.1% to aliquots of the basal medium to provide test media, respectively. After sterilizing these media under heat, the initiator of Lactobacillus casei YIT9026 0.5% was inoculated to the respective medium, and the bacterial strain was cultured at 35 ° C for 5 days to obtain the respective cultures. Each culture was homogenized at 15 MPa, and at 20 parts by weight of that culture, 80 parts by weight of a 15% sugar solution was sterilized at 100 ° C for 5 minutes, and a yogurt flavor was added as well. (product of Yakult Material Co., Ltd.) at 0.1% to prepare a dairy product. A taste test was carried out by 5 highly experienced evaluators in each of the dairy products obtained as described above. No difference was confirmed between the lactic acid bacteria beverages and the control product containing the culture obtained with the use of the basal medium. iii, Additionally, the various extracts were evaluated so that they did not have any effect related to the taste and / or taste of the basal medium and to equalize very well. Therefore, it has also been confirmed that its use in crops for beverages or foods such as beverages of the lactic acid bacteria does not lead to deterioration in its flavors or tastes.

EXAMPLE 7 Effects of the Added Amount of Persimmon Leaf Extract on Taste and Taste and Proliferative Effects (1) Preparation of Persimmon Leaf Extracts Using water and a solution, the pH of which was adjusted to 4.0 with citric acid, in quantities of at most 10 times persimmon leaves, extracts of persimmon leaves were prepared under conditions similar to those of Example 1. These extracts were each concentrated separately at 10 degrees Brix in an evaporator. (2) Determination of a Quantity to be Added Aliquots of a 15% nonfat milk powder medium (with 3% glucose contained therein) was added the persimmon leaf extract prepared in (1) at concentrations above on the scale of 0.01 to 10%, respectively, followed by sterilization at 100 ° C for 60 minutes to prepare a medium for culturing the lactic acid bacteria. To these media, the initiator of Lactobacillus casei YIT9029 at 1% was inoculated, and the strain of bacteria was cultured at 35 ° C until the acidity (titration values of sodium hydroxide to OJ N required for the neutralization of 9g portions). of the respective samples) became 24. The cell count of the lactic acid bacteria in each of the cultures was determined by the BCP medium. Each culture was homogenized at 15 MPa, and at 20 parts by weight of the homogenized culture, 80 parts by weight of a 15% sugar solution was added, which has been sterilized at 100 ° C for 5 minutes, and a flavoring was added as well. of yogurt (product of Yakult Material Co., Ltd.) at 0.1% to prepare a dairy product. With respect to these dairy products, a flavor and taste evaluation was carried out by five trained organoleptic evaluators based on the following standards. The results are shown in Table 4.

Standards of Evaluation (Classification) (Description) A: Very good B: Good C: Average D: Poor E: Very poor TABLE 4 It has been confirmed from Table 4 that the addition of an extract of persimmon leaves at 0.1% or similar is effective for the promotion of the culture of lactic acid bacteria, and additionally, can increase the count of viable cells of the bacterium of lactic acid. It has also been asserted that the addition of an extract of persimmon leaves even when much of 10% to a medium can not provide any additional excellent effect in proportion to the quantities added, but on the contrary, the flavor and taste derived from the extract tend to affect the taste and taste of the product. It has also been confirmed that the growth-promoting effects of the extracts are more markedly exhibited with one obtained by extraction with acid than with one obtained by extraction with water.

EXAMPLE 8 Preparation of Extract 3 The rice husk (mixture of pericarp, aleurone layers and grain germs (brown rice) available from Oryza satvia without the outer layer of the unhatched rice), turmeric (the curcuma of Curcuma longa L), the aerial part of the grass of Houttuvnia cordata Thunb. layers of Eucommia ulmoides Oliv., leaves of persimmon (leaves of Diospyros kaki Thunb.), leaves of Perilla frutescens (L.) Britton var. acuta Kudo, clove (the cocoon of Syzgium aramaticum (L.) Merr. and Perry) and cinnamon (the bark of Cinnamomum zeylanium Nees) were each subjected separately to processing such as debarking and crushing, and then extracted for 60 minutes with hot water at 80 ° C (in an amount of at most 10 times the weight of the corresponding raw material) to prepare extracts of turmeric Houttunynia cordata Thunb., Eucommia ulmoides Oliv., persimmon leaves, knob, clove and cinnamon, respectively. The extracts were each concentrated separately at 10 degrees Brix in an evaporator.

EXAMPLE 9 Determination of Lactic Acid Bacteria Cell Count at the End of the Cultivation (1) Aliquots of a 15% skimmed milk powder medium (with 3% glucose contained in them) as a basal medium, rice husk extracts, persimmon leaves, perilla, Houttunynia cordata Thunb., Eucommia ulmoides Thunb., turmeric, clove and cinnamon, were added. and adjusted to 10 degrees Brix in Example 8, at 1%, respectively, followed by sterilization at 100 ° C for 60 minutes to prepare the sterilized medium. To these sterilized media, the Lactobacillus casei initiator YIT9029 at 1% was inoculated, and the bacterial strain was cultured at 37 ° C until the pH of the respective media reached 3.7. Viable cell counts were determined similarly at the end of the culture. Additionally, a medium with added sodium oleate was also prepared and added in place of the above described extract to give a concentration of 25 ppm in terms of oleic acid and another medium with both of the aforementioned extracts and sodium oleate. The viable cell count was determined similarly at the end of the culture. It should be noted that the determination of each viable cell count was developed by counting colonies formed after incubation of the corresponding sample, which was suitably diluted in a physiological solution, at ° C for 3 days in a BCP medium. The results are shown in the Table TABLE 5 It has been indicated from Table 5 that the combined use of any of the extracts of rice husk, persimmon leaves, perilla, Houttunvnia cordata Thunb., Eucommia ulmoides Thunb., Turmeric, clove and cinnamon, with sodium oleate can synergistically increase the cell count of the lactic acid bacteria at the end of the culture compared to the use of the same extract or sodium oleate alone.

EXAMPLE 10 Determination of Viable Cell Count of Lactic Acid Bacteria in a Dairy Product (1) The cultures prepared in Example 9 (Comparative Products 1 and 2 and Products of Invention 3 and 4) were each separately homogenized at 15 MPa. Aliquots (20 parts by weight) of those homogenized cultures were added 80 parts by weight of a 15% sugar solution that was sterilized at 100 ° C for 5 minutes, followed by the subsequent addition of a 0.1% yoghurt flavoring. to prepare dairy products. These dairy products were filled into containers, and the viable cell count was determined in a similar manner as in Example 9 immediately after the production of the dairy products and after storage at 10 ° C for 14 days. The results are shown in Table 6. iúík? .-. i TABLE 6 It has been indicated from Table 6 that a dairy product obtained by using, as a raw material, a culture prepared by the use of an extract of persimmon leaves and sodium oleate in combination is excellent in its effects to suppress changes in the cell count of the lactic acid bacteria in the product during storage as compared to a milk product available from the use of a culture that does not contain any of them (not added) or that contains only one of them.

EXAMPLE 11 Determination of the Viable Cell Count of Lactic Acid Bacteria upon Finishing the Cultivation (2) Lactobacillus casei YIT9029 was cultured under similar conditions! to those of Example 9 except that aliquots of the basal medium prepared in! In Example 9, various oleate-based emulsifiers were added, respectively, each in combination with 1% persimmon leaf extract prepared in Example 8, so that the emulsifier was quantified at 25 ppm in terms of acid content oleic Viable cell counts of the bacteria in the resulting cultures were determined according to the method of Example 9. The results are shown in Table 7.

TABLE 7 It has been indicated from Table 7 that the use of the oleic acid derived from any of the emulsifiers can provide the resulting culture with a high cell count of the lactic acid bacteria due to the use of persimmon leaf extract in combination. The use of glyceryl oleate, hexaglyceryl monooleate or sucrose oleate between these emulsifiers can provide remarkable effects.

EXAMPLE 12 Determination of Viable Cell Count of Lactic Acid Bacteria at Term of Cultivation (3) Under conditions similar to those of the preparation of the extract in Example 8 except for the use of water and aqueous solutions, the pH of which were adjusted to pH 3.0, 4.0 and 5.0, respectively with citric acid instead of hot water, rice husk, persimmon leaves, Eucommia ulmoides Thunb .. turmeric and clove were each treated to prepare their respective extracts at 10 degrees Brix. To aliquots of a 15% skimmed milk powder medium with the thus prepared extracts added to 0.1%, respectively, sodium oleate at 25 ppm was added in terms of the oleic acid content, and in addition, the Lactobacillus initiator was inoculated casei YIT9029 at 1%. The strain of bacteria was then cultured at 37 ° C until a pH of 3.7 was reached. The viable cell count of the lactic acid bacteria in the resulting cultures was determined according to the method of Example 9. The results are shown in Table 8.

TABLE 8 As indicated from Table 8 that an extract obtained by the acid extraction tends to provide a higher cell count at the end of the culture while the pH of the solvent used in the extraction becomes lower. This effect is observed in a pronounced manner especially with various extracts obtained at pH of 5.0 or less, more preferably, pH of 4.0 or less.

EXAMPLE 13 Determination of the Viable Cell Count of the Lactic Acid Bacteria at the End of the Cultivation (4) Using a citric acid solution of pH 4.0, an extract of 10-degree Brix persimmon leaves was prepared under conditions similar to those of Example 8. To a 10% skimmed milk powder, the 1% extract was added and in addition , sodium oleate was also added at 25 ppm in terms of oleic acid. The resulting mixture was sterilized to prepare a sterilized medium. The aliquots of this sterilized medium were inoculated with various initiators of the 0.1% lactic acid bacteria, respectively, and the bacterial strains were cultured at 37 ° C for 24 hours. As the lactic acid bacteria, Lactobacillus bulgaricus YIT0098, Lactobacillus acidophilus YIT0071 and Lactobacillus casei YIT9029 were used. Additionally, these lactic acid bacteria were cultured in a similar manner as described above in a 10% skimmed milk powder medium for comparison purposes. The cell count of the lactic acid bacteria in the resulting cultures was determined in a manner similar to that of Example 9. The results are shown in Table 9.

TABLE 9 It has been indicated from Table 9 that the effect of increasing the cell count of the lactic acid bacteria, which is available from the combined use of an extract of persimmon leaves extracted with acid and sodium oleate, can be recognized for all lactic acid bacteria although the effect has been confirmed that it somehow differs depending on the species of the lactic acid bacteria.

EXAMPLE 14 Determination of Viable Cell Count of Lactic Acid Bacteria at Term of Cultivation (5) Using a citric acid solution of pH 4.0, an extract of 10 degree Brix persimmon leaves was prepared under conditions similar to those of Example 8. The extract and glyceryl oleate, such as oleic acid, were added to aliquots of a powder medium. of skim milk 15% (with 3% glucose contained in them) so that their added amounts reached those shown below in Table 10, respectively. The resulting medium was sterilized at 100 ° C for 60 minutes, to prepare a sterilized medium. The initiator of Lactobacillus casei YIT9029 at 1% was inoculated into the respective sterilized medium, and the bacterial strain was cultured at 37 ° C until its pH reached 3.7. Additionally, the culture was performed in a similar manner as a control by adding a yeast extract (product of DIFCO), which is generally known as a culture promoter, at 0.2% to the medium. The cell count of the lactic acid bacteria in the resulting cultures was determined in a similar manner as in Example 9. The results are shown in Table 10.

TABLE 10 It has been confirmed from Table 10 that the effect of increasing the viable cell count is clearly recognized by adding 0.1% or more of the persimmon leaf extract and 5 ppm or more of oleic acid in combination. It has also been indicated that the resulting viable cell count is higher than that viable from the addition of a yeast extract.

EXAMPLE 15 Determination of the Viable Cell Count of Lactic Acid Bacteria in a Dairy Product (2) Using the cultures prepared in Example 14, the dairy products were produced in a manner similar to those of Example 10. A taste and taste evaluation was developed by five organoleptic evaluators trained in these dairy products based on the following standards. The results are shown in Table 11.

Standards of Evaluation (Classification) (Description) A: Very good B: Good C: Average D: Poor Very poor TABLE 11 As indicated from Table 11, when the addition of persimmon leaves extract at 10% to the medium, that is, at 2% per dairy product, affects the flavor and taste of the dairy product regardless of the added amount of oleic acid. and therefore, that this added amount can be considered as the upper limit of acceptable addition. It should be appreciated that even when the amount of addition of the persimmon leaf extract is that amount, the product has a better flavor and taste than that available from the addition of the yeast extract.

EXAMPLE 16 Preparation of Extract 4 Leaves of Rubus suavissimus S. Lee (Rosaceae) were subjected to procedures such as debarking, crushing and roasting, and then extracted for 60 minutes with hot water at 90 ° C (in an amount of at most 10 times the weight of the leaves). Rubus suavissimus S. Lee (Rosaceae)), to prepare an extract of Rubus suavissimus S. Lee (Rosaceae). The resulting extract was concentrated at 10 degrees Brix in an evaporator.

EXAMPLE 17 Verification of the Effects for Lactic Acid Bacteria (1) As a basal medium, 12% skim milk powder was supplied. The extract of Rubus suavissimus S. Lee (Rosaceae), which was prepared and adjusted to 10 degrees Brix in Example 16, was added 0.5% to the basal medium followed by sterilization to prepare a sterilized medium. To that sterilized medium, the initiator of Lactobacillus casei YIT9029 at 1% was inoculated, and the bacterial strain was cultured at 37 ° C for 48 hours. Used as a comparative example was one prepared by adding "MEAST" (trademark for autolysate of brewer's yeast, product of Asahi Food and Healthcare Co., Ltd.) at 0.15% to the basal medium and then sterilizing the medium. The amount of "MEAST" thus added is the upper limit of a scale at which the adverse effects on the taste and taste of the crop are acceptable. The proliferation of the lactic acid bacteria in which the culture was then compared with the acidity of the culture (titration value of caustic soda to OJ N when a portion (9g) of culture was taken and an organic acid in the portion culture was reached, unit: mL as an index). The results are shown in Table 12.

TABLE 12 As it is evident from Table 12, it has been confirmed that the acidity becomes greater in a medium with the extract of Rubus suavissimus S. Lee (Rosaceae) added in it than in a medium with no addition or with "MEAST" added in the same. This indicates that the proliferation of lactic acid bacteria can be promoted by the extract of Rubus suavissimus S. Lee (Rosaceae).

EXAMPLE 18 Verification of the Effects for Lactic Acid Bacteria (2) Under conditions similar to those of the preparation method of the extract in Example 16 except for the use of aqueous solutions (90 ° C), the pHs of which have been adjusted to pH 3.0, 4.0 and 5.0, respectively with citric acid instead of hot water, the leaves of Rubus suavissimus S. Lee (Rosaceae) were treated to prepare extracts of Rubus suavissimus S. Lee (Rosaceae) of 10 degrees Brix. To aliquots of a 15% skimmed milk powder medium (with 3% glucose contained therein), said aliquots containing the extracts thus obtained added thereto at 1%, respectively, the Lactobacillus casei YIT9026 initiator was inoculated 1%. The strain of bacteria was cultured at 35 ° C for 5 days. The acidity of the resulting cultures was measured in a similar manner as in Example 17. The results are shown in Table 13.

TABLE 13 As shown in Table 13, it has been confirmed that the proliferation for the lactic acid bacteria tends to become noticeable when an extract of Rubus suavissimus S. Lee (Rosaceae) is obtained by adjusting the pH of an extraction solvent to 5.0 or less.

EXAMPLE 19 Preparation of Extract 5 Leaves of Rubus suavissimus S. Lee (Rosaceae) were subjected to processing such as debarking, crushing and roasting, and then extracted under conditions similar to those of Example 16 with an aqueous solution of citric acid adjusted to a pH of 4.0 (in an amount 10 times the weight of the leaves of Rubus suavissimus S. Lee (Rosaceae)) to prepare an extract of Rubus suavissimus S. Lee (Rosaceae). The extract thus obtained was concentrated at 10 degrees Brix in an evaporator.

EXAMPLE 20 Verification of the Effects for Lactic Acid Bacteria (3) 16% skim milk powder was supplied as a basal medium, and to the medium, the extract of Rubus suavissimus S. Lee (Rosaceae) adjusted to 10 degrees Brix in Example 19 to 1% to prepare a medium. The aliquots of this medium, initiators of various lactic acid bacteria were inoculated at 0.1%, and the strains of bacteria were cultured at 37 ° C for 48 hours. In the previous culture, Lactobacillus casei was used. Lactobacillus acidophilus. Lactobacillus cremoris, Lactobacillus helveticus. Lactobacillus qasseri. Lactobacillus delbrueckii subsp. bulgaricus Streptococcus thermophilus and Lactococcus lactis subsp. lactis The acidity of the resulting cultures was measured in a similar manner as in Example 17 to compare the proliferation of the various lactic acid bacteria. The results are shown in Table 14.

TABLE 14 As is evident from Table 14, the effects of the extract of Rubus suavissimus S. Lee (Rosaceae) on the proliferation of the various lactic acid bacteria have been confirmed with substantially all strains, although they vary depending on the species of the strains . Furthermore, these proliferative effects have been confirmed to have the tendency to give excellent effects to strains that are not very good at proliferation in the basal medium. This suggests that even though it is difficult for lactic acid bacteria to grow in an animal environment is used, the use of the extract of Rubus suavissimus S. Lee (Rosaceae) makes it possible to easily obtain a fermentation product with a large number of cell counts of bacteria.

EXAMPLE 21 Investigation in the Amount of Extract of Rubus suavissimus S. Lee (Rosaceae) to be Added. (1) Preparation of Extract of Rubus suavissimus S. Lee (Rosaceae) Using an aqueous solution of citric acid the pH of which was adjusted to a pH of 4.0, in an amount of at most the leaves of Rubus suavissimus S. Lee (Rosaceae), an extract of Rubus suavissimus S. Lee (Rosaceae) was prepared under conditions similar to those of Example 16. The extract was then concentrated at 10 degrees Brix in an evaporator. (2) Determination of a Quantity to be Added To aliquots of 15% skimmed milk powder medium (with 3% glucose contained therein), the extract of Rubus suavissimus S. Lee (Rosaceae) of 10 degrees Brix, which was prepared before in (1) was added to concentrations on a scale of 0.01 to 10%, respectively, followed by sterilization at 100 ° C for 60 minutes to prepare a medium for culturing the lactic acid bacteria. To these media, the initiator of Lactobacillus casei YIT9029 at 1% was inoculated, and the bacterial strain was cultured at 35 ° C until the acidity (sodium hydroxide titration values at 0J N required for the neutralization of 9g portions). of respective samples) became 24. The cell count of the lactic acid bacteria in each of the cultures was determined by BCP medium. The culture was homogenized at 15 MPa, and at 20 parts by weight of the homogenized culture, was added 80 parts by weight of a 15% sugar solution, which has been sterilized at 100 ° C for 5 minutes at 100 ° C, and added a yogurt flavor (product of Yakult Material Co., Ltd.) at 0.1% to prepare a dairy product. With respect to these dairy products, a flavor and taste evaluation was developed by five trained organoleptic appraisers based on the following standards. The results are shown in Table 15.

Standards of Evaluation (Classification) (Description) A: Very good B: Good C: Average D: Poor E: Very poor TABLE 15 It has been confirmed from table 15 that the addition to 0.01% or similar of an extract of Rubus suavissimus S. Lee (Rosaceae) can provide proliferative effects for the lactic acid bacteria, and furthermore, can increase the viable cell count of the lactic acid bacteria. It has also been asserted that the addition of an extract of Rubus suavissimus S. Lee (Rosaceae) even though much of 10% does not provide extra effects in proportion to the amount added, but on the contrary, tends to affect the taste and taste of the product. It has also been confirmed that the effects of the extract are more remarkably exhibited with one obtained by acid extraction, than with one obtained by extraction with water.

EXAMPLE 22 Verification of the Effects for Lactic Acid Bacteria To aliquots of a 15% skimmed milk powder medium (with 3% glucose contained therein) as a basal medium, extracts of Rubus suavissimus S. Lee (Rosaceae), which were prepared and adjusted to 10 degrees Brix in Example 16 and Example 19, respectively, they were added to 1%, respectively, followed by sterilization at 100 ° C for 60 minutes to prepare a sterilized medium. To these media, the initiator of Lactobacillus casei YIT9029 at 1%, and the bacterial strain, which was cultured at 37 ° C, were inoculated until the pH of the respective medium reached 3.7. The viable cell count was determined at the end of the culture. Additionally, a medium with sodium oleate was added in place of the above-described extract to give a concentration of 25 ppm in terms of oleic acid and another medium, both of the extracts described above and sodium oleate were also prepared. The viable cell count was determined similarly at the end of the culture. It should be noted that the determination of each viable cell count was developed by counting the colonies formed after the incubation of the corresponding sample, which was suitably diluted in a physiological solution, saline at 37 ° C for 3 days in a BCP medium. results are shown in Table 16.

TABLE 16 As confirmed from Table 16 that the combined use of any of the extracts of Rubus suavissimus S. Lee (Rosaceae) with sodium oleate can synergistically increase the cell count of the lactic acid bacteria compared to the use of only the extract of Rubus suavissimus S. Lee (Rosaceae) corresponding.

EXAMPLE 23 Verification of the Effects for Lactic Acid Bacteria (5) The fermentation products of the lactic acid bacterium prepared in Example 22 (Products of the Invention 42, 43, 44 and 45) were each separately homogenized at 15 MPa, and at aliquots of 20 parts by weight of the homogenized products, Aliquots of 80 parts by weight of a 15% sugar solution were added, which was sterilized at 100 ° C for 5 minutes, and a 0.1% yoghurt flavoring was added to prepare dairy products. These dairy products were filled into containers, respectively, and the viable cell count of the lactic acid bacteria in the respective milk products was determined in a similar manner as in Example 22 immediately after its preparation and after storage at 10 °. C for 14 days. The results are shown in Table 17.

TABLE 17 It has been indicated from Table 17 that a dairy product obtained by using, as a raw material, a fermentation product of the lactic acid bacterium, which has been prepared by using only extract of Rubus suavissimus S. Lee (Rosaceae), or an extract of Rubus suavissimus S. Lee (Rosaceae) and sodium oleate in combination, is excellent in the effect of suppressing changes in the cell count of the lactic acid bacteria in the product during storage compared to a dairy product obtained by the use of a fermentation product of the lactic acid bacteria prepared without any of them. In addition, the use of the extract of Rubus suavissimus S. Lee (Rosaceae) in combination with sodium oleate can synergistically contribute the opposite effects to the use of the extract of Rubus suavissimus S. Lee (Rosaceae) alone.

EXAMPLE 24 Verification of the Effects for Lactic Acid Bacteria (6) Using an aqueous solution of citric acid of pH 4.0, an extract of Rubus suavissimus S. Lee (Rosaceae) of 10 degrees Brix was prepared under conditions similar to those of Example 16. To the 10% skim milk powder, the extract was added at 1% and in addition, sodium oleate at 25 ppm was also added in terms of oleic acid. The resulting mixture was sterilized to prepare a sterilized medium. Aliquots of this medium were inoculated with initiators of various 0.1% lactic acid bacteria, respectively, and the strains of the bacteria were cultured at 37 ° C for 24 hours. Like the lactic acid bacteria, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus acidophilis and Lactobacillus casei. As comparative examples, those lactic acid bacteria were also cultured in a similar manner as described above by the use of 10% skimmed milk powder as a medium. The cell count of the lactic acid bacteria in the resulting cultures was determined in a similar manner as in Example 22. The results are shown in Table 18.

It has been indicated from Table 18 that the effects of the extract of Rubus suavissimus S. Lee (Rosaceae) and oleic acid can be recognized by all lactic acid bacteria, although the effects have been confirmed to be somewhat different depending on the species of the lactic acid bacteria.

EXAMPLE 25 Verification of the Effects for Lactic Acid Bacteria (7) To aliquots of a 15% nonfat milk powder medium (with 3% glucose contained therein) as a basal medium, various oleate-based emulsifiers were added respectively, each in combination with Rubus suavissimus S. extract. Lee (Rosaceae) al 1% prepared and adjusted to 10 degrees Brix in Example 19, so that the emulsifiers quantified at 25 ppm in terms of the content of the oleic acid. The resulting mixtures were then sterilized at 100 ° C for 60 minutes to prepare a sterilized medium, respectively. To these media, the initiator of Lactobacillus casei YIT9029 at 1% was inoculated, and the bacterial strain was cultured at 37 ° C until the pH of the respective medium reached 3.7. The viable cell count was measured in a manner similar to Example 21. The results are shown in Table 19.

TABLE 19 It has been confirmed that as shown in Table 19, the use of oleic acid as a derivative of any of the emulsifiers can provide proliferative effects for the lactic acid bacterium due to the use of the extract of Rubus suavissimus S. Lee (Rosaceae) in combination . The use of sodium oleate monoglyceryl oleate or sucrose oleate between these emulsifiers can provide remarkable effects.

EXAMPLE 26 Verification of the Effects for Lactic Acid Bacteria (8) Using an aqueous solution of citric acid of pH 4.0, an extract of Rubus suavissimus S. Lee (Rosaceae) adjusted to 10 degrees Brix was prepared under conditions similar to those of Example 16. The extract and glyceryl oleate, such as oleic acid, were added to aliquots of 15% skim milk powder medium (with 3% glucose contained therein) so that the amount added reached those shown in Table 20, respectively. The resulting medium was sterilized at 100 ° C for 60 minutes to prepare a sterilized medium. To the respective sterilized media, the initiator of Lactobacillus casei YIT9029 at 1% was inoculated, and the bacterial strain was cultured at 37 ° C until its pH reached 3.7. Additionally, the culture was performed in a similar manner as a control by adding a yeast extract (product of DIFCO), which is generally known as a culture promoter, at 0.2% to the medium. The cell count of the lactic acid bacteria in the resulting cultures was determined in a manner similar to Example 22. The results are shown in Table 20.

TABLE 20 As shown in Table 20, the proliferative effects for lactic acid bacteria can be recognized by the addition of oleic acid at 0.01 ppm or higher.

EXAMPLE 27 Verification of the Effects for Lactic Acid Bacteria (9) Using the lactic acid fermentation product prepared in Example 26, dairy products were produced in a manner similar to that of Example 21. Taste evaluations and tests were developed by five organoleptic evaluators trained in those dairy products based on evaluation standards. similar to those of Example 21. The results are shown in Table 21.

TABLE 21 It has been found from Table 21 and that similarly to that indicated in Table 15, the addition of the extract of Rubus suavissimus S. Lee (Rosaceae) at 10% to the medium, that is, at 2% per product, affects the taste and taste of the products regardless of the added amount of oleic acid. It should be noted that even with the addition of the extract of Rubus suavissimus S. Lee (Rosaceae) in that amount, the products had a better taste and taste than those available from the addition of the yeast extract.

INDUSTRIAL APPLICATION The fermentation product of the lactic acid bacteria of the present invention has a large number of viable cell counts of the lactic acid bacteria. The fermentation product does not suffer much deterioration in flavor and taste because the death of the lactic acid bacteria can be reduced. Accordingly, this fermentation product of the lactic acid bacteria can be suitably used as a raw material for various fermented dairy foods.

Claims (13)

NOVELTY OF THE INVENTION CLAIMS

1. - A fermentation product of the lactic acid bacterium, which has been obtained by culturing the lactic acid bacteria in a medium comprising an extract of at least one food material selected from the group consisting of rice husk, leaves of persimmon, perilla, Houttunyia cordata Thunb, Eucommia ulmoides Oliv .. turmeric, clove, cinnamon and Rubus suavissimus S. Lee (Rosaceae).

2. The fermentation product according to claim 1, further characterized in that the extract has been obtained by acid extraction.

3. The fermentation product according to claim 1, further characterized in that the extract has been obtained by acid extraction under an acidic condition not greater than a pH of 4.0.

4. The fermentation product according to any of claims 1 to 3, further characterized in that the amount of the extract is equivalent to a scale of 0.01 to 10% by weight.

5. A fermentation product of the lactic acid bacterium, which has been obtained by culturing the lactic acid bacteria in a medium comprising an extract of at least one selected food material IíUá of the group consisting of rice husk, persimmon leaves, goatee, Houttunyia cordata Thunb, Eucommia ulmoides Oliv., Turmeric, clove, cinnamon and Rubus suavissimus S. Lee (Rosaceae), and oleic acid or a derivative thereof.

6. The fermentation product according to claim 5, further characterized in that the oleic acid or the derivative thereof is an ester of oleic acid selected from the group consisting of glyceryl oleate, polyglyceryl oleate ester, and sucrose oleate, or a metal salt of oleic acid.

7. The fermentation product according to claim 5 or 6, further characterized in that the oleic acid or derivative thereof is equivalent to a scale of 1 to 50 ppm.

8. The fermentation product according to claim 5, further characterized in that the extract has been obtained by acid extraction.

9. The fermentation product according to claim 5, further characterized in that the extract has been obtained by acid extraction under an acid condition not greater than pH of 4.0 or less.

10. The fermentation product according to any of claims 5 to 9, further characterized in that the extract is equivalent to a scale of 0.01 to 10% by weight.

11. - A fermented milk food comprising a fermentation product of any of claims 1 to 10.

12. A method for the production of a fermentation product of the lactic acid bacterium, which comprises culturing the lactic acid bacteria in a medium comprising an extract of at least one food material selected from the group consisting of rice husk, persimmon leaves, perilla, Houttunyia cordata Thunb. Eucommia ulmoides Oliv., Turmeric, clove, cinnamon and Rubus suavissimus S. Lee (Rosaceae).

13. A method for the production of a fermentation product of the lactic acid bacterium, which comprises culturing the lactic acid bacterium in a medium comprising an extract of at least one food material selected from the group consisting of a lactic acid shell. rice, persimmon leaves, goatee, Houttunyia cordata Thunb. Eucommia ulmoides Oliv., Turmeric, clove, cinnamon and Rubus suavíssimus S. Lee (Rosaceae), and oleic acid or a derivative thereof.