JPS60141281A - Method and apparatus for producing granule of microorganism - Google Patents

Abstract

PURPOSE:To obtain dried bacterial cell having excellent handleability and high preservability, by mixing living bacterial cells with a solution for forming a protective film, pouring the obtained mixture into a coagulation salt solution, separating the obtained coagulum, and drying the product. CONSTITUTION:For example, 5-10pts. of bacterial cells such as cells of bifidus bacteria are mixed with a protective film-forming solution containing 10-50pts. of starch and 0.1-1.0pt. of sodium alginate, and the mixture is poured in a mass into a neutral salt solution for coagulation composed of calcium lactate, calcium chloride, etc. to effect the coagulation of the mixture. The obtained coagulum is taken out of the solution and dried. Preferably, the product is coated with an oil or fat having a melting point of higher than the body temperature to prevent the extermination of the bacteria with gastric juice.

Description

[Detailed Description of the Invention] and an apparatus, and a method for producing granular bacterial cells that are less likely to be killed by gastric juice and reach the intestine even after oral ingestion, especially in the case of Bifidus 1 cells. The aim is to provide equipment suitable for this purpose.

BACKGROUND ART Conventionally, when preserving the entirety of many microorganisms such as bacteria and yeast, methods have been used to dry and preserve the microorganisms.

For drying, the bacterial cells isolated from the culture solution can be dried as is, or starch, lactose, sugar, or organic or inorganic salts can be added to them.
Moisture is reduced to 10% by low temperature ventilation drying, vacuum drying or freeze drying.
1. For example, Bifidobacterium, which is thought to be lacking in the adult intestines, can be ingested raw by mixing it with drinks or yogurt, but it can be dried from places that are not suitable for storage and mixed with other foods. This method is widely used, and Japanese Patent Publication No. 58-46293 discloses that a base formulation for tableting containing one or more of starch, starch hydrolyzate or protein with a moisture content of 4% or less and freeze-dried A method for producing a tablet confectionery containing bifidus by mixing 1 bifidus and tableting is disclosed, and Japanese Patent Application Laid-Open No. 58-149
Publication No. 67 discloses a method for increasing the survival rate of Bifidobacteria by mixing Bifidobacterial cells and raw starch and freeze-drying the mixture.

Dried bifidobacteria produced by the method mentioned above have a considerable shelf life, but they are basically in powder form and have a large contact area with air, so special consideration is required for preservation. When taken into the body, it is dispersed uniformly and is affected by gastric juices, killing a considerable number of bacteria, and it has drawbacks such as a small percentage reaching the intestines.

Furthermore, it is well known that even if lactic acid bacteria, yeast, etc. are molded by granulation, extrusion, etc. and dried by known methods, there are many problems with storage stability.

The present inventors conducted research to fundamentally solve the drawbacks of the above drying method and obtain a drying box that is easy to handle and has good storage stability. Focusing on the fact that it is better to suspend the bacterial cells in a protective film-forming solution instead of letting them dry and then react with a coagulating salt solution to solidify and form a uniform protective film on the surface of the bacterial cells, we first The body and the protective film forming solution are uniformly mixed, and the mixture is poured into a saline solution for total coagulation to coagulate into a lump.The obtained coagulated product is separated and dried, and if necessary, the melting point is lower than body temperature. They succeeded in developing a method for coating with oils and fats that have a high viscosity, as well as a device suitable for the process.

Microorganisms used in the present invention include milk I-1m1 yeast and other microorganisms used for food, drug A production or processing, or general industrial use. suitable. Examples of bifidobacteria include Bifidobacterium intes (Bifidobacterium intes).
l]acterium 1nfantis), Bifidobacterium longum (Bifidobacterium longum)
um longum), Bifidobacterium adl.
ecsentis).

The above-mentioned 1. spheroids are cultured in a natural or synthetic culture medium containing sugars, salts, and growth factors, and after culturing, the box is usually separated and concentrated using a centrifuge, and water or physiological saline is added to the concentrate. Dilute, wash, and centrifuge again. The above-mentioned washing operation may be repeated 2 to 3 times depending on the degree of concentration of the bacterial cells. The obtained washed image body has the culture medium components sufficiently removed and becomes a bacterial cell with little off-flavor, so it can be used as is, or may be further dehydrated in some cases.

In addition, the protective film forming solution mixed with the bacterial cells contains a substance such as sodium alginate, potassium alginate, pectin, or glucomannan that forms a protective film on the surface of the bacterial cells by binding with Gimpo ions such as calcime ions. It contains starch, dextrin, sucrose, monosodium glutamate,
It is recommended to add a substance that has a full humidity control function such as sodium ascorbate, completely prevents sudden dehydration shock during drying, and prevents oxidation. Examples of the composition of such a mixture include, for example, 5 to 10 parts of bifidus as a solid;
Starch 10-50 parts, sodium alginate 01-10
part, and 05-20 parts of sodium glutamate and 05-10 parts of sodium ascorbate! l'! rA
Examples include liquids.

The coagulating salt solution used is a substance that has the function of combining with the main component of the protective film-forming solution to form a film on its surface, and in practical terms it does not kill bacterial cells and is harmless to the human body. Neutral 5 such as calcium lactate and calcium chloride
A solution of the order of 1% at ~25°C is used.

Bifidobacterium longum (Bifidba)
cterium longum)-2 in the above-mentioned manner and producing granules of 4 pieces of bif (1 bird)-fcm will be explained with reference to the attached drawings. In the figure, 1 shows the entire raw material mixing tank, inside which is equipped with m stirring VjP and 2, bifidobacterial cells, sodium alginate, starch, sodium ascorbate, and sodium glutamate are added in the proportions shown and mixed uniformly. At this time, the viscosity of mixture A is usually about 200 CI). The resulting mixture A is then transferred via pump 3 to mixture storage tank 4 .

The storage tank 4 is of a closed type, and the upper part thereof is provided with a pipe 6 communicating with an air compressor 5 as a pressure regulating device and a pressure regulating extension 6', so that an appropriate pressure, e.g. An extrusion pressure of 3 kg/- is applied to the storage tank 4.

Further, four stirrers 7 are provided at the bottom of the storage tank 4 to prevent starch and the like from settling. Further, a pipe U8 is connected to a desired position of the storage tank 4, and the other end of the pipe 8 is opened to a large number of distribution pipes 9. A large number of nozzles 10 are attached to the distribution pipe 9 facing downward, and the inner diameter of the nozzles 10 is preferably one thread or less, particularly in the range of 0.2 to 0.5 mm. Further, it is preferable that each nozzle 10 is provided with all the pulp 11 and that the injection can be stopped after fully adjusting the flow rate.

According to the above configuration, the inside of the storage tank 4 is pressurized by the air compressor 5, so the mixture A is injected from the tip of the nozzle 10, but the amount of injection is controlled by the air compressor 5.
The pressure can be adjusted by adjusting the pressure of the air and the valve 11, but instead of the air compressor 5, a pump or other device capable of adjusting the amount of ejection may be provided in the middle of the pipe 8.

A coagulating liquid tank 12 is provided below the nozzle 10, and a solution of calcium lactate or calcium chloride is put therein to form a coagulating liquid B. Further, an overflow gutter 13 is provided on the - side, and the coagulating liquid B can be supplied to the other side via a coagulating liquid storage tank 14 and a metering pump 15, so that the liquid level is approximately 3 to 10 cr below the lower end of the nozzle 10. position.

Therefore, the mixture A from the nozzle 10 is always injected under constant conditions, solidifies in a short period of time in the coagulating liquid tank 12, and settles in granular form. At this time, since the solids in mixture A are uniformly suspended in the sodium alginate solution, the particles first come into contact with the calcium ions, instantly forming a complete film, and gradually the calcium ions penetrate inside the body. A film is formed and the whole solidifies.

The extrusion pressure from the nozzle 10 is preferably adjusted depending on the diameter of the nozzle 10, the composition of the mixture, and the distance between the nozzle 10 and the liquid surface.
Granules of any desired size, from fine granules to large granules, can be produced by controlling the amount.

If the extrusion pressure is too strong and the distance between the nozzle and the liquid level is insufficient, the mixture A cannot be made into granules but becomes continuous rod-shaped, so care must be taken. In this way, mixture A
become coagulated particles C that do not stick together and settle downward.

In order to take out the coagulated particles C2, a screen 1.6 of 24 to 32 meshes is run in a direction perpendicular to the flow of the coagulated liquid B, and the coagulation tank 12 is surrounded by pulleys 17, 1.7... and the screen 16 is discharged. A gutter 18 is provided on the side so that water can be sprayed from a nozzle 19 provided above for cleaning. As a result, the coagulated particles C are washed, and when the screen 6 rotates, the direction changes downward and falls into the container 20, but the coagulated particles C still attached are washed away by the screen 16.
(removed by the airflow from the air blowing nozzle 21 provided on the back side of n11, and all collected into the container 20.

The recovered coagulated particles C contain a large amount of water and have poor preservability if left as is, so they are then dried. For drying, a vacuum drying method, a freeze-drying method, a drying method using an inert gas, etc. can be used, and the moisture content after drying is preferably about 1 to 5 grams.

The granules obtained by the method described in 2 have a structure in which the oil and fat layer in Fig. 3 are excluded, and the surfaces of the granules (a) and starch (b) are coated with a film of alginated calcium (c). It has a structure in which the diameter is swollen to 2 to 3 times the inner diameter of the nozzle 10, but as can be seen from the figure, (as in the conventional case, the bacterial cells and drying aid are mixed together; Since it is complete and uniform, and the coating/zero layer is extremely thin, it has a significantly improved storage capacity, and is easy to handle because it is in a granular form.

Next, the integral granules obtained in Example 1 were wrapped in aluminum foil and stored at room temperature and 37°C.
Show C.

Table 1 The granules obtained by the method of the present invention can be taken orally as is, mixed with other foods, and further used for processing other products or for industrial purposes. It is something. After oral extraction, the kudzu body is covered with alginic liquid Cal/Um, so it is unlikely to be killed by gastric juice, but
In order to further reduce mortality, it is desirable to coat the box granules with whole oil and fat. The oil and fat used is a low-melting point oil that is conventionally used for coating modified materials, but cannot be reused, and a unique high-1'X oil that does not dissolve at human body temperature.
It is a hydrogenated oil with an I rating, for example 37°C to 43°C.

Coating with oil or fat can be carried out by a conventional method, but it is convenient to use one fluidized bed granulation coating device.

The device sends gas from below to fluidize the granules and spray oil and fat7. This coating method significantly improves resistance to gastric juices, and most of the bacteria reach the intestines.

An example of this will now be shown with an experimental example.

The experiment was carried out using a full fluidized bed granulation coating device Flow Coater pLo-5 model (trade name) produced by the method of Example 1, and 2 kg of bifidobacteria granules.
! 11, modified 40℃ hardened oil 8002 i, i at 45℃
! The test was carried out by spraying into an air stream at 50°C.

As shown in Figure 3, the resulting coated product becomes a stag with a layer of bacterial cells (a) and starch (b) on the surface of the granules (oil and fat layer). (containing 0.2 parts of common salt and 0.3 parts of pepsin, adjusted to pH 1.5 with hydrochloric acid) and reacted with stirring in a constant temperature bath at 37°C for 1 to 5 hours. Anti-1; +Tt
After adjusting the pH to 7.0, the bifidus viability was measured by a conventional method. The results are shown in Table 2.

Table 2 However, the bifidus solid number is 17 granules.

In fact, it is safe to say that the number of living cabinets obtained as above is due to fluctuations within experimental error and there is virtually no dead number of rtc jj at all.

Furthermore, when it reaches the small intestine, it is digested by the digestive juices (into an oily film) and the granules also disintegrate, so there is some degree of death due to the influence of the digestive juices. In tests conducted by the present inventors using artificial intestinal fluid, approximately 20% of the Bifidobacteria were killed, so most of the Bifidobacteria can reach the large intestine alive.

Although the above experiment was conducted on bifidus, the method of the present invention can be applied to the drying of many prophylactic organisms, including lactic acid bacteria and yeast. In addition, since it is coated uniformly, it will not lose its shape even if it is stored as is or added to neutral or acidic foods with high moisture content, and it is less affected by the moisture in the food, making it easier to use than conventional It is possible to maintain the activity of the yarn a2 for a long period of time compared to the dried product.

This will be explained below using examples.

Example 1 Hifidobacterium longum (BlfIdbac
The tooth bodies were cultured anaerobically at 37° C. for 18 hours in a skimmed milk medium digested with T. terium Iongum) i thanasinase (trade name), and after the culture, the tooth bodies were concentrated using a centrifuge. Add physiological saline to the concentrated solution to the original volume, centrifuge it again, wash it, repeat the washing twice, and culture it. f)
I got k.

To 700 tons of the raw saliva, 2002 tons of potato starch, 2 Of monosodium glutamate, and 101 tons of sodium L-ascorbate were added, and 200 me'' of a 25% sodium alginate solution dissolved in advance was added and the whole was mixed uniformly.

The above-mentioned liquid mixture was continuously extruded using the apparatus shown in FIG. 1 and coagulated into granules. This was scooped out with a screen of 32 mesioles of gold = i, l)3, washed with water and sufficiently dehydrated, spread on a metal tray to a thickness of 5 to 10 degrees and freeze-dried. The average particle size of the obtained granules is approximately Q,
9 mm, specific solubility) f# 2.75 nr13
/ ? The yield was 3,402 ml of white granules with a water content of 20 g.

The above bifidobacteria granules contain 5.8 x 10 bifidobacteria.
It contained 8/7 and maintained its activity for a long time when stored in a sealed container, making it suitable as an additive to foods such as yogurt.

Example 2 Milk whey protease decomposition product 1.0%'! p including z
Using H6,2 medium, Streptococcus thermophilus 74 (St, thermophilus
) i4Q°C for 4 hours, centrifuged and washed in the same manner as in Example 1, yielding about 7,002 sludge cells per 101 medium (
Approximately 1501' as a solid! &Obtained.

The above bacterial cells were mixed with a protective film forming solution in the same manner as in Example 1, granulated and freeze-dried to give an average particle size of about 10 mm and a specific volume of 2.
．． 75ml/y, water 2.0% white granules 3507
I got it.

The above-mentioned granules had a bacterial count of 2 x 10"Bi!#. After packaging with aluminum foil that is not breathable or moisture permeable, the granules were subjected to a storage test, and the results shown in Table 3 were obtained.

Table 3 Example 3 Bifidobacterium Iongum obtained by the method of Example 1
) for 700 recitations, Ma Lingcao starch 4007, monosodium glutamate 207, and sodium L-ascorbate 10! i', 5% rhomethoxyl pectin 10
0#C was added and mixed uniformly.

The above mixed solution was prepared in the same manner as in Example 1, and the inner diameter was 0.4. mm
The granules were injected from a height of 51+ into a 1st part calcium lactate solution from a nozzle, scooped out with a 32 sieve screen, washed with running water, thoroughly dehydrated, freeze-dried, and crushed to an average particle size of 1 mm by specific volume. White granules 5302 with a water content of .0 ml/f and a moisture content of 20 parts were obtained. The number of bifidus strokes in this granule is 2. OX], 081 hardness/・2, and is suitable for long-term storage.

Example 4 Zanocalomyces cerevisociae (Sac, Ce! l
ev i cae) f Aerated culture at 30°C in cane molasses medium, centrifuged and washed to reach a concentration of about 70
0flO live cocoons were obtained.

The above bacterial cells were mixed with corn starch 200F and 5% rhomethoxyl pectin 10001111, extruded from a height of 7 cm from a nozzle with an inner diameter of 0.5 m + π in the same manner as in Example 1, granulated, and scooped out with a 32 mesh screen. After that, it was washed with light and water, and then truly dried to obtain an average particle size of 1.2.
Approximately 4.50 f'e of granules with +i1 m x 50% moisture were obtained. These granules did not lose their fermentation ability even after long-term medical treatment.

[Brief explanation of the drawing]

FIG. 1 is an explanatory diagram of the apparatus, FIG. 2 is a cross-sectional view of the coagulating liquid tank of FIG. 1, and FIG. 3 is an enlarged view of one part of the coated granules. 1...Mixing tank 4 Storage tank 6.8...Piping 10-nozzle] 2...Coagulation liquid tank 14...Coagulation liquid storage tank 16...
Screen 20 ・Container A ・Mixture B ・Coagulation liquid C...Coagulation particle patent applicant Meiji Dairies Co., Ltd.

Claims (4)

[Claims] (1) Mix viable bacterial cells and a protective film-forming solution, inject the mixed solution into a saline solution for total coagulation to coagulate, take out the coagulated product, dry it, and if necessary, heat it to a temperature above body temperature. A method for producing solid granules, characterized by coating them with an oil or fat having a melting point. (2) The method for producing bacterial granules according to claim 1, wherein the living box is made of bifidobacteria. (3) The mixture of the living box and the protective film forming solution contains 5 to 10 parts of bacterial cells, 10 to 50 parts of starch, and 01 to 1.0 parts of sodium alginate in terms of dry weight ratio. The method for producing bacterial cell granules according to Scope 1. (4) Living cocoon and protection: a storage tank that stores all of the protective film forming solution, a nozzle that communicates with the storage tank and hangs downward, a coagulation tank that is provided so that the liquid level is located below the nozzle, and Depending on the device for adjusting the jetting amount of the nozzle, the nozzle has an inner diameter of 1 mm or less, preferably 0.2 to 0.5 m, and the liquid level distance between the tip of the nozzle and the coagulation tank is about 3 to 10 CTL. A box granule manufacturing device characterized by: