JPH06133735A - Enteric-coated lactobacillus granule - Google Patents

Abstract

PURPOSE:To obtain the subject granules insoluble in the mouth and stomach but soluble in the intestinal tract, thus excellent in lactobacterial proliferation and contamination resistance by coating lactobacillis and coffee leaves or extract thereof with specific two kinds of layers. CONSTITUTION:The objective granules can be obtained by coating lactobacillis such as Lactobacillus casei or Leuconostoc dextranicum (viable cell number: >=10<6>) and coffee leaves or extract thereof with (A) a layer containing fatty oil (e.g. palm oil, coconut oil) and vehicle (potato starch, egg, glucose) and (B) a layer containing hydrous alcohol-soluble protein (pref. zein, a glyceral fatty acid ester is used as plasticizer).

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention contains a lactic acid bacterium growth factor excellent in the effect of promoting the growth of useful general lactic acid bacteria and lactic acid bacteria such as bifidobacteria that live in the human intestine or are contained in foods. The present invention relates to an enteric-coated lactic acid bacterium granulated product, and more particularly to an enteric-coated lactic acid bacterium granulated product which is not dissolved in the stomach after ingestion through the mouth but is first dissolved in the intestinal tract and is excellent in growth and activation of lactic acid bacterium in the intestinal tract.

[0002]

2. Description of the Related Art Generally, 100 trillion bacteria are inhabited in the human intestine, and it is said that there are more than 400 kinds of bacteria. (Science of Bifidobacteria. Mio Maeda. P8
Of the various intestinal bacterial flora that have recently been composed, such as general lactic acid bacteria (for example, Lactobacillus, Leucon).
Enteroactive bacteria such as ostoc, Streptococcus and Bifidobacterium lactic acid bacteria not only produce metabolites harmful to the host (human) but also suppress the growth of harmful bacteria that produce them. Has the effect of

Therefore, by paying attention to such properties, intestinal active bacteria are added to foods and drinks and tablets, and by ingesting them, the intestinal bacterial flora is predominantly changed to the bacterial flora. However, attempts have been made to help prevent and treat various diseases of the digestive system. However, general lactic acid bacteria and bifidobacteria are cultivated under conditions of pH 4.5 or higher, anaerobic, and 36 to 42 ° C., and particularly in the presence of dissolved oxygen at pH 4.2 or lower and high temperature of 45 ° C. or higher, Die to. Therefore, if general lactic acid bacteria and bifidobacteria are simply ingested directly from the mouth, gastric juice (pH 1.0 to 1.5, H
It is killed by 0.2 to 0.5% by weight of Cl (hereinafter referred to as "%"), and the utilization efficiency in the intestinal tract is significantly impaired.

Conventionally, for example, JP-A-60-255731.
There has been proposed a method for producing bifidobacterial granules as described in Japanese Patent Laid-Open Publication No. This method is a method in which the pressure damage during granulation is alleviated and the survival rate of the bifidobacteria is increased by granulating the bifidobacteria with granules such as saccharides, organic acids, and hardened oil. However, in this method, although the survival rate of the bifidobacteria in the granules is increased, the bifidobacteria are killed when the granules are treated with artificial gastric juice (pH 1.2). Therefore, when the above-mentioned granules are taken from the mouth, a considerable number of cells are killed by the gastric juice, and the proportion of the cells remaining as viable cells in the intestine decreases.
Further, since the bacterial cells thus granulated have a large contact area with the air, a special preservation method for maintaining the activity of the bacterial cells is required.

As a method for solving the above-mentioned drawbacks, Japanese Patent Laid-Open No.
The method disclosed in Japanese Unexamined Patent Publication No. 0-141281 can be mentioned. In this method, a protective film-forming solution such as sodium alginate and starch is mixed with viable cells, and the mixture is dropped into a coagulating solution using a pore nozzle having an inner diameter of 1 mm or less, coagulated, dried, and then coated with oil and fat. It is a thing. However, in this method, a viable cell-containing solution tank, a pore nozzle for dropping it, a coagulation tank for coagulating a viable cell-containing solution, and an apparatus for washing, collecting and drying after coagulation A special equipment tank is required. In addition, it takes a long time for production because the amount of dropping of each nozzle is small,
It has the drawback of being difficult to mass-produce.

On the other hand, oligosaccharides, lactulose, ginseng extracts, nitrogen sources, vitamins and the like are known as growth or growth promoting factors for lactic acid bacteria. However, even if these growth promoting factors were ingested together with lactic acid bacteria, it was impaired by gastric juice and the growth and growth effects of lactic acid bacteria in the intestinal tract could not be expected so much.

[0007]

SUMMARY OF THE INVENTION The present invention has been made in view of the above circumstances, and its purpose is to obtain lactic acid bacteria for the first time in the intestinal tract after ingestion without being dissolved in the stomach. By dissolving, it can be eluted in a large amount in the intestinal tract without being killed under the influence of the pH of gastric juice, and can promote the growth and growth of lactic acid bacteria in the intestinal tract. It is also to provide excellent enteric-coated lactic acid bacterium granules.

[0008]

Means for Solving the Problems The above-mentioned object is characterized in that an lactic acid bacterium and a coffee leaf or an extract thereof are coated with two layers shown in (A) and (B) below. Achieved by lactic acid bacteria granulation. (A) Oil-and-fat and excipient-containing layer (B) Hydrous alcohol-soluble protein-containing layer.

That is, the present inventors have found an enteric-coated granulated product which can be dissolved by the intestinal digestive fluid without being dissolved in the gastric juice after ingestion and can promote the activation of lactic acid bacteria in the intestinal tract. Study was carried out. As a result, lactic acid bacteria and coffee leaves or their extracts are (A) a layer containing fats and oils and a excipient (long-term storage stabilizing layer), and (B) a hydrous alcohol-soluble protein-containing layer (saliva or gastric juice). When the granulation product is composed of two layers, the dissolution prevention layer) and the dissolution in the gastric juice can be prevented, and it can be dissolved in the intestinal tract for the first time. The active ingredient of coffee leaves promotes the growth and growth of lactic acid bacteria. They have found that the physiological activity of lactic acid bacteria can be maximized, and that they can be granulated products that have no contact with air during long-term storage and have excellent storage stability, and have reached the present invention.

Next, the present invention will be described in detail. The lactic acid bacteria referred to in the present invention include general lactic acid bacteria, bifidobacteria and the like which are useful in the human intestine. Examples of general lactic acid bacteria include Lactobacillus (hereinafter referred to as “L.”) casei, L. et al. bulgaric
us, L. plant, Leuconostoc
(Hereinafter referred to as "Leu.") Dextranicu
m, Leu. citrovorum, Streptoc
occus thermophilus, L. faec
alis, L .; acidophilus etc. are mentioned. In addition, as Bifidobacterium, Bifidobac
terium (hereinafter referred to as "Bif.") bifid
um, Buf. adolescentis, Bif. l
ongum and the like.

These lactic acid bacteria may be used alone or in combination of several kinds. It is preferable that the viable cell count of these lactic acid bacteria is 10 ⁶ cells / g or more from the viewpoint of activation in the intestinal tract.
For these lactic acid bacteria, it is preferable to use freeze-dried bacterial cell powder.

Next, as the coffee leaf, a young leaf of a plant of the genus Coffea, which is widely cultivated in tropical and subtropical regions, is used as a raw material of coffee. As the above-mentioned young leaves, it is sufficient to use young leaves such as Arabica and Robusta which are cultivated species which are easily available, alone or in combination of several kinds. Among these, new shoots that are cut off as waste sprouts during coffee cultivation are preferable, and those having a length of 4 to 10 cm or less and a width of 1 to 3 cm or less on the branch tip side are preferable.

As a form to be used, fresh coffee leaves just washed with water, or fresh coffee leaves after washing with water, and then 1
Up to 2 weeks of air-dried coffee leaves, hot-air dried, roasted, freeze-dried coffee leaves or the like, or an extract obtained by extracting the active ingredient from the coffee leaves with water or another solvent, or a powder or paste thereof. And the like, and the like can be selected as appropriate. The coffee leaf contains a component that promotes the growth of lactic acid bacteria. By incorporating the coffee leaf or its extract in the granulated product together with the lactic acid bacteria, the growth of the lactic acid bacteria in the intestine is promoted. The amount of coffee leaves used is preferably 0.1% by weight (hereinafter referred to as “%”) or more in terms of solid content in the total weight of the enteric-coated lactic acid bacterium granules. If it is less than 0.1%, the effect on the growth and growth of lactic acid bacteria tends to be low.

Of the two layers covering the lactic acid bacterium and coffee leaves or extracts thereof (hereinafter, these two are collectively referred to as "granulated product"), the granulated product is stabilized for a long period of time. For (A) layer (hereinafter referred to as “A layer”) for
Oils and fats and excipients are used. In the present invention, the term “coating” also includes the case where the granules are mixed in the coating layer. First, as fats and oils, vegetable fats and oils such as palm oil, palm oil, soybean oil, rapeseed oil, and cacao butter and solid fats such as hardened oils obtained by curing them, rice wax, candelilla wax, edible wax such as honey wax, etc. Etc.
The melting point of these oils and fats is 30 to 4 in terms of uniform dispersibility and spreadability in a temperature range that does not impair the effectiveness of the granulated material.
5 ° C is preferred.

Similarly, the excipients used in the layer A include starch, egg, starch, protein, glucose, lactose, sucrose, starch and eggs made from potato, corn, rice, wheat and the like. Examples include sugars such as maltose, which may be used alone or in combination of several kinds.

Next, of the two layers covering the above-mentioned granules, the (B) layer (hereinafter referred to as "B layer") for preventing the granules from dissolving in saliva or gastric juice is A hydrous alcohol-soluble protein is used. Examples of the hydrous alcohol-soluble protein include zein contained in corn, and vegetable or animal proteins derived from wheat, soybean, rice, collagen, gelatin and the like. These may be used alone or in combination of two or more. Further, for example, after treating the zein with an alkali, a fraction such as a zein peptide having a molecular weight of 5,000 to 40,000 obtained by extracting with acetone is used, or the untreated zein and the zein peptide are used in combination. May be. Of these, the use of zein is preferable because it becomes more insoluble. In addition, about 10% of a water-soluble protein may be mixed with the hydrous alcohol-soluble protein to be used.

In addition, in order to enhance the uniform dissolution and dispersibility of the hydrous alcohol-soluble protein in the B layer, if necessary,
It is better to use a plasticizer. Examples of the plasticizer include emulsifiers such as glycerin fatty acid ester, sucrose fatty acid ester, polyglycerin fatty acid ester, and sorbitan fatty acid ester, and glycerin and sugar alcohol. Among these, glycerin fatty acid ester is particularly preferable in terms of uniform dissolution and dispersibility of hydrous alcohol-soluble protein and uniform coverage when the granules are coated. The amount of the plasticizer added varies depending on the amount of the hydrous alcohol-soluble protein, but is preferably about 0.8% of the total weight of the layer B solution.

As for the coating of the above-mentioned A layer and B layer on the material to be granulated, either the A layer or the B layer may be coated first. When the layer A is first coated and then the layer B is coated, the dissolution resistance is more excellent, and the work efficiency in producing the granulated product is good. On the contrary, when the B layer is first coated and then the A layer is coated, a granulated product having a smaller particle size can be obtained.
Further, the enteric-coated granulated product of the present invention may be formed as a multi-layered granulated product by alternately coating and forming several layers A and B.

The enteric-coated enteric-active-bacterium granules of the present invention are produced, for example, as follows. That is, in the case of coating the layer A first, first, the granulated substance and the excipient are mixed,
To this is added oil and fat which has been melted in a liquid state in advance, and mixed and stirred while keeping the temperature. Insulation temperature is uniform dispersibility of fats and oils,
Depending on the type of the granulated material, the temperature is preferably 45 ° C. or lower, more preferably 35 to 45 ° C. or lower from the viewpoint of preventing thermal denaturation.

The amount of the oil / fat used is preferably 10 to 30% of the total weight of the layer A. If the fat and oil is less than 10%, the surface of the granulated product cannot be sufficiently covered with the fat and oil, and the long-term storage stability and the dissolution resistance tend to be poor.
On the other hand, if it exceeds 30%, a slip phenomenon occurs during granulation, and granulation tends to be difficult. The amount of the excipient used may be set appropriately, but from the viewpoints of uniform dispersibility of fats and oils, suitability for coating fats and oils on granules, and suitability for granulation, the amount is 30 to 8 in the total weight of the layer A.
It is preferably 0%.

Next, the above mixture is supplied to a granulating device while being stirred, and granulated to obtain an A layer-coated granulated product. Examples of the granulating device include a granulator with a screen, an extruder, and the like, and a device provided with a cooling means is preferable because the granulated product can be solidified quickly. It is desirable that the A layer-coated granulated product is left to stand for a few hours in advance at a temperature of 20 ° C. or lower and aged before the subsequent coating of the B layer. If this stationary aging is insufficient, the A layer granules are likely to be bound together or softened during the subsequent B layer coating granulation, and the granulation suitability tends to deteriorate.

The layer A-coated granules thus obtained are coated with layer B by spraying, dipping or the like. That is,
First, a hydrous alcohol-soluble protein is dispersed and dissolved in hydrous alcohol. The hydrous alcohol used here preferably has an alcohol concentration of about 85 to 95%. That is,
Outside this range, the uniform dispersion and solubility of the hydrous alcohol-soluble protein in the hydrous alcohol tend to deteriorate. At this time, the ratio of the hydrous alcohol-soluble protein to the hydrous alcohol is preferably 6 to 14 hydrous alcohol to 1 hydrous alcohol-soluble protein in terms of uniform solubility and dispersibility. At this time, a plasticizer is added as needed.

Next, the step of drying while applying the solution dispersed and dissolved in the hydrous alcohol to the surface of the granules coated with the layer A is repeated to form the layer B. Examples of the application method include spraying and dipping. For example, when spraying,
A rolling machine such as a revolving pan, a fluidized dryer, a centrifugal fluidized granulation dryer, or the like may be used.

The hydroalcoholic soluble protein in the total weight of the granulated product thus obtained is preferably 5 to 5
It is desirable that the content is 0%, more preferably 15 to 35%. If the content of the hydroalcoholic protein is less than 5%, the solubility resistance tends to deteriorate, and if it exceeds 50%, the mouthfeel tends to deteriorate.

The enteric-coated lactic acid bacterium granules thus obtained may be taken as they are, or the granules may be tableted or formulated. Alternatively, foods such as health foods may be prepared by adding flavor components such as flavors and dairy products.

When the layer A is first coated, the layer A may contain the hydrous alcohol-soluble protein used in the layer B. It takes a long time to form the (B) layer because the application of the hydrous alcohol-soluble protein and the drying are repeated, but it is necessary to mix the hydrous alcohol-soluble protein in the (A) layer as well. Since the number of layers formed can be reduced, the time can be shortened and the enteric effect is high. Further, an oil / fat layer may be provided between the A layer and the B layer, or a sugar coating layer may be provided on the surface of the granule.

[0027]

As described above, the granulated product of the present invention comprises an A layer (long-term storage stability stabilizing layer) containing fats and oils and a shaping agent,
Since the granules are coated with the two layers of the layer B (dissolution-resistant layer) containing the hydrous alcohol-soluble protein, the granules are not dissolved by the gastric juice and are dissolved in the intestine for the first time. Therefore, the lactic acid bacterium and the active ingredient of the coffee leaf that grows and proliferates the lactic acid bacterium can exert its efficacy in the intestinal tract without being affected by the gastric juice after being orally ingested.

Further, even when stored in a usual packaged state, the effect of the active ingredient is not lowered, and the long-term stability is excellent. Moreover, since it is almost tasteless and odorless, it is a granulated product excellent in versatility without impairing the flavor of the food itself even when used in food such as health food. Also, the granulated product is
Since it can be continuously manufactured by using a conventional granulating device or the like, it can be mass-produced.

The present invention will be specifically described below with reference to examples. [Example 1] Lactobacillus acidophilus (L.a.
Dry powder of lactic acid bacteria (cidophilus) (live cell count 1
0 ¹⁰ / g) 27 parts by weight (hereinafter referred to as "parts"), coffee Hayu extract FD products (raw coffee leaves 150 parts 9
What was extracted with 3000 parts of 0 ° C. hot water for 1 hour was made into a vacuum concentrate, which was further subjected to FD. ) 8 parts, 170 parts of skim milk powder as an excipient and 28 parts of glucose are mixed, 75 parts of palm oil and fat dissolved at 40 ° C. are added to this mixture, and the mixture is stirred and then extruded with a screen having a pore diameter of 0.8 mm. With a granulator, an A layer-coated granulated product having a length of 1.5 to 2 mm was obtained.

Next, 75 parts of zein was dispersed and dissolved in 488 parts of an aqueous ethanol solution (275 parts of ethanol, 113 parts of water) little by little, and then 4 parts of glycerin fatty acid ester was added to obtain a coating solution. Then, using a centrifugal granulation dryer, the above-mentioned A was performed under the conditions of a rotor rotation speed of 120 rpm, a product temperature of 20 ° C. and a blower of 150 L / min.
521 parts of the above coating solution was sprayed on the layer-coated granules and dried to obtain enteric-coated lactic acid bacterium granules.

Comparative Example 1 A granulated product was obtained in the same manner as in Example 1 except that the coating solution was not used.

[Comparative Example 2] A granulated product was obtained in the same manner as in Example 1 except that palm oil was replaced with water.

1.00 g of the enteric-coated lactic acid bacterium granules obtained in the above-mentioned Example 1 and Comparative Examples 1 and 2 were weighed out, and artificial gastric juice (0.
1M-HCl, NaCl 12%, pepsin 0.32%,
It is immersed in 50 g of pH 1.2) for 3 hours, then filtered, and the obtained residue is washed with sterilized water, and immediately after that, artificial intestinal fluid (0.1M-KH ₂ PO ₄ , 0.1M-Na ₂ ) is added. HPO ₄
-It was immersed in 2H ₂ O, 1: 6, pH 7.5) at 37 ° C for 2 hours. The number of remaining viable lactic acid bacteria after the immersion was examined by a conventional plate culture method. As the initial viable cell count, the viable cell count after the immersion of only the artificial intestinal fluid (without the immersion of the artificial gastric juice) was examined. Furthermore,
Each granulated product was stored at 20 ° C. for 4 months, and then the viable cell count was examined. Further, in Example 1, after immersing in artificial intestinal fluid for 2 hours, 0.1 cc was sampled and inoculated into GYP medium,
The cells were cultivated aerobically at 24 ° C. for 24 hours, and the multiplication factor of lactic acid bacteria was examined on the basis of the number of non-additive coffee leaves. The results are shown in Table 1.
Shown in.

[0034]

[Table 1]

From the above results, the enteric-coated granules of the present invention are excellent in gastric juice resistance and the solubility of lactic acid bacteria in the intestinal juice,
It was also a granulated product excellent in the growth of lactic acid bacteria.

[Examples 2 to 5] Granules were obtained in the same manner as in Example 1 except that the amount of fats and oils (% by weight in the total weight of the layer A coated granules) was changed to the ratio shown in Table 2. It was The gastric juice resistance and the multiplication factor of lactic acid bacteria of the obtained granules were examined in the same manner as in Example 1. The amount of skim milk powder was adjusted as the amount of fat and oil increased. The above results are shown in Table 2.

[0037]

[Table 2]

From the results shown in Table 2, the number of remaining viable bacteria was high even with the addition of 30% of oil and fat, the slipping phenomenon did not occur during granulation, and the granulation suitability was good. In addition, the ability of lactic acid bacteria to grow in intestinal fluid was also excellent.

[Examples 6 to 8] Granules were prepared in the same manner as in Example 1 except that the zein of Example 1 was replaced with each hydrous alcohol-soluble protein fractionated so as to be soluble in hydrous alcohol as shown in Table 3. After preparation, the number of remaining viable bacteria was measured. The results are shown in Table 3.

[0040]

[Table 3]

From the results shown in Table 3, the enteric-coated lactic acid bacterium granules of the present invention using the hydrous alcohol-soluble protein were granules excellent in gastric juice resistance.

[Examples 9 to 13] Granules were prepared in the same manner as in Example 1 except that the zein content in Example 1 was changed as shown in Table 4, and the residual viable cell count was measured. The results are also shown in Table 4.

[0043]

[Table 4]

From the results shown in Table 4, the zein content is 15%.
Above all, the granulated product exhibited good dissolution resistance and was good.

Example 14 The procedure of Example 1 was repeated, except that the coating solution of Example 1 was combined with zein and gelatin as follows. 8 parts of gelatin was dissolved in 113 parts of water, and zein 7 was added to a solvent solution containing 375 parts of ethanol.
5 parts by little was added little by little to disperse and dissolve, and then 4 parts of sugar alcohol was added to obtain a coating solution. The number of remaining viable bacteria of the obtained enteric-coated lactic acid bacterium granule was as good as 10 ⁸ cells / g, which was good.

[Example 15] Lactobacillus L. acidophilus 27 parts of dried lactic acid bacterial powder (10 ¹⁰ viable cells / g) and coffee leaf powder 12
A coating solution in which 75 parts of zein was dispersed and dissolved in 488 parts of an ethanol aqueous solution (375 parts of ethanol, 113 parts of water) was coated and dried by a centrifugal drying granulator. Next, 100 parts of skimmed milk powder, 50 parts of powdered sugar and 15 parts of hardened palm oil and fat melted
The mixture with 0 parts was coated on the coated dried product using a rotary kettle to obtain an enteric-coated lactic acid bacterium granule. The obtained enteric-coated lactic acid bacterium granules had an initial viable cell number of 10 ⁸ / g, a residual viable cell number of 10 ⁸ / g, a multiplication factor of 14.8 after culturing, and gastric juice resistance,
The growth of lactic acid bacteria was good.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁵ Identification code Internal reference number FI Technical display area A61K 35/78 C 7167-4C 47/14 D 7433-4C 47/42 D 7433-4C C12N 1 / 04 7236-4B

Claims (1)

[Claims] 1. An enteric-coated lactic acid bacterium granulated product comprising a lactic acid bacterium and a coffee leaf or an extract thereof, which are coated with two layers shown in the following (A) and (B). (A) Oil-and-fat and excipient-containing layer (B) Hydrous alcohol-soluble protein-containing layer.