JP2596418B2 - Useful microorganism-containing granule and method for producing the same - Google Patents

Description

Description: FIELD OF THE INVENTION The present invention relates to a useful microorganism-containing granule which is less killed during granulation and has improved storage stability, and a method for producing the same.

[Prior Art] Useful microorganisms such as yeast and bacteria can be widely used in foods such as dairy products and brewed products, lactic acid bacteria preparations, pharmaceuticals such as intestinal medicines, and microbial pesticides.

Microorganisms are generally vulnerable to heat and are easily deactivated in the presence of water.However, they can be stored for a long time under dry conditions. , Protein, or organic or inorganic salts, and dried at low temperature through aeration, vacuum drying or freeze-drying to prepare cells having a water content of 10% or less or cell-containing powder.

Since the dry cells obtained in this manner are powder, (i) the surface area is large, and therefore the contact area with air is large, so that it is easy to absorb moisture, and there is a problem in handling in an open system. (Ii) ) Due to its large hygroscopic property, the cells can die if care is taken not to block the moisture during long-term storage. (Iii) Since the powder is in the form of powder, the powder will flutter during handling and contaminate the environment. Causing bridging during industrial handling, causing trouble; (iv) easy to become powder and difficult to handle when added to food, etc .; In the case of powder, it is inconvenient to take out the whole amount, and there are problems such as:

[Problems to be Solved by the Invention] Therefore, various methods for granulating microbial cell powder into granules that are easy to handle have been proposed.

For example, starch and a starch hydrolyzate having a water content of 4% or less, one or two or more basic blends for tableting of protein and a freeze-dried bifidobacterium are mixed, and the mixture is tabletted to give a bifidobacterium-containing tablet confection. Production method (Japanese Patent Publication No. 58-46293)
No.), a method of mixing cells and raw starch and freeze-drying (Japanese Patent Application Laid-Open No. 58-14967), a method of encapsulating the cells in a seaweed gel and producing a cell-containing particle gel (Japanese Patent Application Laid-Open No. 57-186446). No.), lactic acid bacteria obtained by suction filtration of lactic acid bacteria and chlorella dry powder as a filtration layer were dissolved in a 10% gelatin solution, then sprayed into chlorella dry powder, and then a 50-60 # granular lactic acid bacteria-chlorella mixture was obtained. Production method (JP-A-53-99371), a method of mixing viable cells and a protective film-forming solution, injecting into a salt solution for coagulation, gelling and covering, and granulating (JP-A-60-141281) However, in all of these methods, except for the tableting method, granulation is carried out in an aqueous system, so that the mortality of the bacterial cells during handling is large and the efficiency is low.

That is, when granulated in an aqueous system, (1) since the binder uses a water-soluble substance, it is hydrophilic and has high water absorption and hygroscopicity after granulation, and (2) finally evaporates water. Therefore, it is difficult to apply to bacteria that are sensitive to water and heat, and (3) pores are formed in the granules due to the evaporation of water, the specific surface area is large, and moisture is easily absorbed. .

Also in the tableting method, there is a problem that storage stability is poor because the surface area of the powder cannot be essentially improved. In order to solve these problems, it is conceivable to granulate with a non-aqueous system, but as a method of granulating with a non-aqueous system, a fluid granulation method in which a core component is coated with a binder and a cell component is proposed. However, according to this method, only a granule having a small number of bacteria per unit weight can be obtained due to the presence of a core material in the center of the granule. , It is easily affected by humidity, storage stability is not sufficient, and its commercial value is low.

In addition, since the core material is used, only those with a large particle size can be made,
It is difficult to make things less than 1.0mmφ.

Accordingly, an object of the present invention is to solve the above-mentioned problems in useful microorganism-containing granules, and to suppress inactivation as much as possible when granulating cells or a cell-containing material powder, maintain appropriate strength, and store, An object of the present invention is to provide a useful microorganism-containing granule which is hardly crushed during transportation and use, has low hygroscopicity, and has little deactivation during storage, and a method for producing the same.

[Means for Solving the Problems] The present invention is based on the total weight of the granule composition while heating and melting a binder substance having a melting point of 35 to 100 ° C. to a powder of useful microbial cells or a substance thereof. A method for producing useful microorganism-containing granules, characterized in that the granules are added in the range of 10 to 50% by weight and are stirred and granulated in a non-aqueous system, and the cells obtained by the method are substantially uniform in the granules. The present invention provides a useful microorganism-containing granule characterized in that the granule is contained in a granule.

Microorganisms to be granulated in the present invention may be any cells, but the characteristics of the present invention are particularly useful in the case of cells that are vulnerable to heat and moisture and have difficulty in storage.

For example, Bacilles, M. Pusill, which produces various proteases (low-temperature active API-21, rennet, etc.) and useful enzymes such as lipase, amylase, cellulase, pectinase, etc.
us, Asp.niger, etc., various bacteria and yeasts (Sacch.ce) used in the brewing field such as sake, miso, soy sauce, natto, etc.
revisiae, Asp.Oryzae, Asp.tamarii, Sacch.rouxii, Bac.n
Atto, Bac. Subtillus, Asp. sojae, Acetobacter, etc.), lactic acid bacteria and bifidobacteria Micococcus genus, Streptococcus
Genus, Pediococcus, Leuconotoc, Lactobacillus,
Bacteria belonging to the genus Bifidobacterium are exemplified.

These cells are separated from the culture solution and dried as they are under low-temperature air-drying, vacuum-dried or freeze-dried, or powdered cells containing starch, lactose, proteins or salts added to the separated cells and dried in the same manner. Is used.

There are various methods of granulating in a non-aqueous system, and there is a method of granulating by dissolving a binder wax in an organic solvent,
In this method, as in the case of the aqueous system, the pore rate is large, the physical properties of the granules are not improved, and the influence of the cells on the solvent is large. In comparison with this, it was found that this method of granulating by heating at a temperature higher than the melting point of the binder without using a solvent had a pore rate as small as 0.1 cc / g or less and was the best without the influence of the solvent.

As the binder substance at the time of granulation, the following wax-like substances are usually used.

(1) Animal or vegetable waxes such as beef tallow, rice wax, wood wax, beeswax, candelilla wax, carnauba wax, lanolin and other natural fats and oils, (2) sucrose esters, glycerin fatty acid mono (di (3) synthetic polymers having wax-like properties as in the above (1) and (2), such as cellulose derivatives: benzylamino, etc., tri) esters, propylene glycol fatty acid (PG) esters, sorbitan fatty acid esters, etc. Methylcellulose, dimethylaminomethylcellulose, piperidylethylhydroxyethylcellulose, cellulose acetate diethylaminoacetate, cellulose acetate dibutylaminohydroxypropyl ether, etc .; polyvinyl derivative: vinyl diethylamine-vinyl acetate copolymer , Vinylbenzylamine-vinyl acetate copolymer, polyvinyldiethylaminoacetoacetal, vinylpiperidylacetoacetal-vinylacetate copolymer, polyvinylacetal diethylaminoacetate, polydimethylaminoethyl methacrylate, polydiethylaminomethylstyrene, polyvinylethylpyridine, vinylethylpyridine-styrene copolymer, Vinylethylpyridine-
Acrylonitrile copolymer, methylvinylpyridine-
Acrylonitrile copolymer, methylvinylpyridine-
Styrene copolymer and the like.

The binder material used in the present invention has a melting point of 100 ° C. or less, preferably 40 to 60 ° C., and among those that do not affect the cells, those that are optimal according to the ultimate purpose of use. Select

For example, when used as a food additive, such as lactic acid bacteria and bifidobacteria, it is necessary to select from those permitted for food use, such as sucrose esters, glycerin fatty acid esters, PG esters, and sorbitan fatty acid esters. Or one selected from natural oils and fats
Mixtures of more than one species are used.

To keep the granules of the final product stable for a long time,
Since the smaller the hygroscopicity, the more stable the wax material should be selected because the ratio of the hydrophobic portion is large. From this viewpoint, for example, high fatty acid sucrose ester, sorbitan high fatty acid ester, rice wax and the like are preferable.

In the present invention, an additive such as a bulking agent for keeping the number of viable bacteria in the granules constant can be used at the time of granulation, if desired. Such additives do not affect the cells,
What is suitable for the final purpose, for example, starch, protein, glucose, lactose, inorganic salts (such as Na ₂ SO ₄ ), and organic salts are appropriately used.

Since the amount of the binder substance (wax) to be added greatly depends on the physical properties of the cells to be granulated, the optimum amount of addition must be determined for each case. Is appropriate, and preferably 15 to 30% by weight. If it is less than 10% by weight, granulation is almost impossible, while if it is more than 50% by weight, granulation at the time of granulation is quick, the granulation efficiency is poor, and the physical properties of the granules are also unfavorable.

In the present invention, the above-mentioned dried cells or the cell-containing powder, the binder substance and, if desired, other additives are granulated in a non-aqueous system.

Rolling granulation, fluidized granulation, extrusion granulation,
Spray granulation, stirring granulation, etc. are conceivable. (1) In rolling granulation, the core material is used as described above, so there are problems with the bacterial concentration, particle size, storage stability, and (2) fluidized granulation. In the grains, the pore rate is large, the strength is weak,
Others have the same problems as (1). (3) Extrusion granulation requires a large amount of wax, large pore ratio and small shape cannot be used. (4) Spray granulation dissolves. It is difficult to uniformly disperse the wax in the wax, it takes a long time to disperse the wax, and the apparatus is large.

In comparison, it has been found that only the stirring granulation method without using a solvent can overcome these disadvantages.

As the granulator, a granulator having a structure which can be heated by hot water, steam, electric heat or the like so as to be maintained at a temperature not lower than the melting point of the wax to be used is used.

As a method of adding the wax, any of a method of adding the powder to the granulator in the form of powder at the same time as the cell powder to be granulated, and a method of adding a wax previously melted at the time of granulation can be granulated,
In order to granulate in as short a time as possible and to prevent the death of the cells, it is preferable to add a molten wax in a spray state.

The temperature at the time of granulation is a temperature higher than the melting point of the wax-like substance used as the binder, preferably in the range of the melting point + 5 ° C. Granules can be obtained.

[Effects of the Invention] According to the present invention, (1) since granulation is performed at a low temperature in a non-aqueous system, there is no inactivation due to the action of water and heat during granulation. (2) The obtained granules are It has a structure that contains useful microorganisms substantially uniformly in the binder, so that it is possible to produce granules with a high number of bacteria, and that it has high strength. (3) The shape is nearly spherical, and the flowability is remarkably good. (4) Since the binder wax forms a layer on the surface, there are few pores on the surface, the hygroscopicity is improved, and there is little deactivation during storage. (5) Granulation due to its nearly spherical shape. (6) that the binder used for granulation or other binder used in the final step can be easily coated, a structure having a dense skin on the surface can be obtained, and the storage stability can be further improved. Because of its spherical shape and excellent fluidity, granules can be used for other products. When mixed with water, there is no possibility of powdering or bridging, and the handling is significantly improved. (7) Since no solvent (water or organic solvent) is used, solvent removal is unnecessary. Excellent effects such as simple process can be obtained.

[Examples] Hereinafter, the present invention will be described with reference to Examples and Comparative Examples.

Examples 1 to 5 A lactic acid bacterium belonging to the genus Lactobacillus was diluted with starch, lactic acid, or the like, and a lactic acid bacterium-containing material prepared at a cell amount of 3 × 10 ¹⁰ cells / g was mixed with a binder (sucrose ester) having a ratio shown in Table 1. −
Monoglycerides and rice waxes) and bulking agents (Na
₂ SO ₄ ) was added and granulated at a temperature of 50 to 70 ° C. (Examples 1 to 5), and the viable cell remaining ratio was measured by a conventional method to obtain the results shown in Table 1. When the granules obtained in Examples 1, 2, 4, and 5 were stored at room temperature and 35 ° C., and the viable cell survival rate was measured,
As shown in Table 2, it was clarified that long-term stable storage was possible.

Examples 6 to 8 1 × 10 ⁹ Bifidobacterium belonging to the genus Bifidobacterium
/ g of the bulk powder prepared so as to contain the binder (sucrose ester-sorbitan ester-glycerin ester, rice wax) and a bulking agent (Na ₂ SO ₄ , glucose) in the proportions shown in Table 1. Granulation was performed at a temperature of 55 to 70 ° C., and the viable cell remaining ratio was measured to obtain the results shown in Table 1. When a storage stability test was performed on these three types of granulated products in the same manner as in Examples 1 to 5, good results were obtained.

Comparative Examples 1-2 Examples 1-5 and in the same cell-containing material 20% aqueous solution of glucose and bulking agent (Na ₂ SO ₄₎ and added in the proportions shown in Table 1,
The mixture was stirred and granulated at room temperature, and the viable cell remaining ratio was measured.
As shown in the figure, only extremely low values were obtained.

Continued on the front page (51) Int.Cl. ⁶ Identification number Agency reference number FI Technical indication C12R 1: 225) (C12N 1/20 C12R 1:01)

Claims (5)

(57) [Claims] 1. A method for adding a binder material having a melting point of 35 to 100 ° C. to a powder of useful microbial cells or a substance containing the same in an amount of 10 to 50% by weight based on the total weight of the granule composition. And a method for producing useful microorganism-containing granules, wherein the granules are stirred and granulated in a non-aqueous system. 2. The method according to claim 1, wherein the amount of the powder of the useful microorganism cells or the content thereof is 30% by weight or more based on the total weight of the granule composition. 3. The binder material is a waxy material,
2. The method according to claim 1, wherein the granulation temperature is equal to or higher than the melting point of the wax-like substance. 4. The method according to claim 1, wherein the surface of the granule is coated with a binder used for granulation in the final step of granulation or a different binder. 5. A method for adding a binder material having a melting point of 35 to 100 ° C. to a powder of useful microbial cells or a substance containing the same in an amount of 10 to 50% by weight based on the total weight of the granule composition. And useful microorganism-containing granules obtained by stirring and granulating in a non-aqueous system, wherein the granules substantially uniformly contain bacterial cells in the granules.