JP2003009848A - Microorganism composition and method for degrading organic waste with the composition - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method for efficiently degrading organic wastes, especially biological wastes, particularly a method for degrading the organic wastes, especially the biological wastes, in an extremely short time substantially without by-producing a stink during or after the degradation. SOLUTION: This microorganism composition comprises the following microorganisms: (1) Bacillus subtilis, (2) Bacillus sp. KB-02 (FERM P-18364), (3) Bacillus vallismortis, (4) Bacillus licheniformis, and (5) Bacillus pumilus. And, the method for degrading the organic wastes, especially biological wastes, is characterized by comprising the following processes: (i) a process for mixing the organic wastes, additional nutritive sources for the microorganisms, and the above-described microorganism composition, (ii) a process for heating the mixture obtained in the process (i) at about 40 deg.C to about 65 deg.C, and (iii) a process for keeping the mixture at an average temperature of about 40 to about 65 deg.C.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a method for decomposing and treating organic waste using microorganisms. In particular, the present invention relates to a method for degrading biological wastes including vegetable waste, fish roe, okara, etc. using microorganisms. Further, the present invention is
It relates to a microbial composition suitable for such a decomposition method. Furthermore, the invention relates to degradation products obtained by such a degradation method.

[0002]

2. Description of the Related Art Organic wastes, especially kitchen wastes (so-called garbage) including vegetable scraps and fish tarps, biological wastes including animal and plant residues such as wastes in the agriculture / livestock industry or food processing industry , Is directly disposed of by a method such as incineration or landfill, or is processed by various devices generally called a garbage disposal machine. However, methods such as incineration and landfill are not desirable in the present situation where effective utilization of resources is strongly desired, and in the case of incineration, much energy is required, which is also disadvantageous. On the other hand, a device generally called a food waste disposer is roughly classified into a compost type / disappearing type, a dry type and a carbonizing type. The compost type / annihilation type is a method utilizing fermentation by microorganisms, and the dry type and the carbonization type are methods using a drying furnace or a carbonization furnace. Among them, the dry type and the carbonized type require a considerably high temperature furnace, which is disadvantageous in terms of energy cost, but has an advantage that the processing time is short. On the other hand, it is considered that the composting type / extinguishing type is advantageous in terms of resource reuse and cost, but in general, since fermentation by microorganisms does not proceed rapidly, there is a problem that the decomposition processing speed is relatively slow. It was pointed out. here,
Both the composting method and the extinction method are methods of decomposing organic wastes using microorganisms, and there are cases in which they are argued separately because of differences in discharge frequency and quality of decomposed products, but they are not always strictly distinguished. It's not something you can do. Further, in the compost type / extinguishing type, since organic waste is decomposed by fermentation, the generation of odor during fermentation is a particular problem. In order to suppress the odor during fermentation, various heating methods and times, stirring methods and times, and water content adjustment have been studied, and a number of devices capable of controlling these have been developed and sold.
However, conventionally, in the decomposition of organic wastes using such microorganisms, the emphasis has mainly been on the decomposition of the carbon skeleton of organic compounds, and in general, N ( Nitrogen) and S
Little consideration was given to the behavior of (sulfur).
Therefore, no matter how the temperature, water content, aeration, etc. are controlled, it is difficult to suppress the generation of ammonia, amine, and sulfides such as hydrogen sulfide and sulfur dioxide derived from the decomposition of nitrogen- and sulfur-containing organic compounds. However, it was also an obstacle in terms of utilization of degradation products. Due to such a problem, a device with a deodorizing device added has been developed and sold, but it is hard to say that it has achieved its purpose sufficiently. Also, it has been pointed out that a used bacterial bed containing a decomposition product requires a large number of secondary treatments for reuse or is incinerated as it is. Therefore, there is a demand for a method for decomposing organic waste, which can decompose organic waste efficiently in a shorter time, facilitates recycling of decomposition products, and generates less odor during decomposition and less odor of decomposition products. .

[0003]

SUMMARY OF THE INVENTION It is an object of the present invention to provide an efficient method for decomposing organic waste, especially biological waste. Particularly, it is an object of the present invention to provide a method for decomposing organic wastes, particularly biological wastes in a very short time with little generation of malodor during and after decomposition. Further, it is an object of the present invention to provide a method for decomposing an organic waste and a decomposition product thereof, in which the decomposition product can be effectively used. Moreover, this invention aims at providing the microbial composition suitable for the above-mentioned method.

The present invention is directed to a method of degrading organic wastes, particularly biological wastes, using a mixture of microorganisms, degradation products, and microbial compositions of defined composition for use in the method. It is a thing. The present inventors combine a plurality of specific microorganisms including nitrogen-fixing bacteria and maintain a constant fermentation condition, so that a bad odor is less likely to occur during and after decomposition, and organic wastes, particularly living organisms, can be generated in an extremely short time. It was found that the system waste can be decomposed and the decomposition product can be effectively used, and the present invention has been completed. That is, the microbial composition of the present invention is a microbial composition containing the following microorganisms: (1) Bacillus subtilis (2) Bacillus sp. KB-02 (FERM P-18364) (3) Bacillus varismortis ( 4) Bacillus licheniformis (5) Bacillus pumilus More specifically, the microbial composition of the present invention comprises (1) to
A microbial composition containing all of the microorganisms of (5) and containing each of the microorganisms of (1) to (5) in a proportion of 5% or more with respect to the total amount of the microorganisms of (1) to (5) in the composition. Is. The microbial composition of the present invention is also a microbial composition containing a mixture containing the above-mentioned microorganisms (1) to (5) in approximately equal amounts.
The present invention also includes the above-mentioned microorganisms (1) to (5),
In addition, (6) Paenibacillus sp. KB-06 (FERM P-1836
A microbial composition containing 8). Here, the amount and proportion of microorganisms are based on the number of cells or the mass of cells. Also,
The present invention is a method for decomposing organic waste, particularly biological waste, which comprises the following steps. (I) warming the temperature of the mixture obtained in step (ii) (i) of mixing the organic waste, the additional nutrient source for microorganisms and the above microbial composition to about 40 ° C to about 65 ° C. Step (iii) maintaining an average temperature of about 40 ° C to about 65 ° C. In particular, the invention is characterized by including the following steps:
It is a method of decomposing organic waste, especially biological waste. (I) heating the temperature of the mixture obtained in step (ii) (i) of mixing organic waste, an additional nutrient source for microorganisms, and the above-mentioned microbial composition to about 40 ° C to about 65 ° C. Step (iii) A step of continuously or at regular intervals while maintaining an average temperature of 40 ° C to 65 ° C.

The present invention is also the use of the above-described microbial composition of the present invention for the decomposition of organic wastes, especially biological wastes.

BEST MODE FOR CARRYING OUT THE INVENTION The microorganism contained in the microbial composition of the present invention is Bacillus subtilis.
s), Bacillus vallismo
rtis), Bacillus sp.KB-02 (Bacillus sp.) (FERM P-1
8364), Bacillus lichen
iformis), Bacillus pumilus
And Paenibacillus sp. (Paenibacillus sp.) KB-06
(FERM P-18368).

These are all thermotolerant bacteria. These microorganisms can survive from 0 ° C to at least 65 ° C. Bacillus subtitis is known to produce protease, α-amylase, cellulase, glutamine synthetase that assimilates ammonia, and bacitracin-like antibiotics. Bacillus vallismortis and Bacillus sp.
KB-02 (FERM P-18364) is similar to Bacillus subtilis and is especially viable at high temperatures. Bacillus licheniform
is is known to produce protease, α-amylase, cellulase, glutamine synthetase that assimilates ammonia, and bacitracin-like antibiotics like Bacillus subtitis. Bacillus pumilus is a ruminal bacterium and is known to produce xylan-degrading enzymes such as xylanase and β-xylosidase and lignin-degrading enzymes. Also, Paenibacillus KB-06 (FER
M P-18368) is a high temperature-resistant nitrogen-fixing bacterium. The microbial composition of the present invention may be a combination of microorganisms equivalent to these microorganisms. In addition to these microorganisms, the microbial composition of the present invention may contain other microorganisms and other materials generally used for decomposition of organic waste by the microorganisms. In particular, in addition to the above-mentioned microorganisms to be contained in the composition of the present invention, as long as they do not significantly affect the decomposition rate of organic waste and the quality of decomposition products in terms of generation of ammonia, amine or sulfide, etc. Microorganisms or other materials may also be included. Other materials that may be contained in the microbial composition of the present invention include, for example, bulking agents, preservatives and the like, more specifically, for example, amino acids,
Mineral water containing sugars, vitamins, and trace elements, such as Botanic (Japan Research Institute for Mineral Applications) is included. Microorganisms that may be further included in the microbial composition of the present invention,
Actinomycetes, yeast and others are included. In general, the microbial compositions of the present invention also include media components and microbial metabolites suitable for the growth of those microorganisms.

As examples of these microorganisms suitable for the present invention, various strains deposited on June 4, 2001, at the Patent Organism Depositary Center, National Institute of Advanced Industrial Science and Technology can be used. Table 1 shows the relationship between the flybate name and the accession number given by the inventors for each strain. These private names are also used herein. Among the bacteria constituting the microbial composition of the present invention, Bacillus sp. KB-02 and Paenibacillus sp. K
Bacteria other than B-06 are available from various other institutions or research facilities, and also commercially available. Alternatively, it can be isolated from the natural world, and the isolated bacterium may be identified according to various documents with reference to growth temperature, sugar catabolism, staining property, 16S rRNA and the like. Therefore, the Bacillus
Regarding microorganisms other than KB-02 and Paenibacillus sp. KB-06, the microorganisms are limited to the corresponding strains deposited even when referred to in the present specification as "microorganism of (1)". Make it not exist.

[Table 1] Table 1. Relationship between the private name of the deposited microorganism and the deposit number

In particular, Bacillus subtilis KB-01 (which is a Bacillus subtilis commonly found in nature) and Bacillus sp. KB-02 have the following mycological properties.

[0008]

[Table 2] Table 2. Bacillus subtilis KB-01 and Bacillus
The following mycological properties of sp. KB-02

[0009]

[Table 3] Table 2. Continued

In addition to the above, the colonies of KB-01 are flower-shaped, whereas the colonies of KB-02 are circular and KB-02 is
Protease activity is stronger than -01.

The microbial composition of the present invention is (1) Bacillus subtilis (2) Bacillus Balismortis (3) Bacillus sp. KB-02 (4) Bacillus licheniformis (5) Bacillus pumilus Or further (6) Paenibacillus sp. KB-06 A microbial composition comprising:

The composition of the microbial composition of the present invention can vary considerably. Generally, the above (1)-
(1) in the composition containing all of the microorganisms of (5)
About 5% or more of each of the microorganisms (1) to (5) relative to the total amount of the microorganisms (5), more preferably (1) in the composition.
It is preferable that each of the microorganisms (1) to (5) is contained in an amount of about 10% or more based on the total amount of the microorganisms (5) to (5). Most preferably, the microbial composition of the present invention comprises about (1) to (5) all of each of the microorganisms relative to the total amount of these microorganisms in the composition.
It is an almost equivalent mixture containing 20% (total amount is 100%). Furthermore, the microbial composition of the present invention comprises (1) to
The microorganism of (5) may be contained, and further the microorganism of (6), which is a nitrogen-fixing bacterium, may be contained. When the microbial composition of the present invention contains (6) Paenibacillus sp. KB-06 (FERM P-18368), it contains all of the microorganisms of (1) to (6) and (1) to (6) in the composition. ) To the total amount of microorganisms, (1) ~
It is preferable that the microorganisms of (6) each contain about 5% or more,
More preferably, each of the microorganisms (1) to (6) contains about 10% or more of the total amount of the microorganisms (1) to (6) in the composition. Most preferably, the microbial composition of the present invention is a nearly equimolar mixture of microorganisms of (1)-(6).

The microbial composition of the present invention is prepared by growing each microorganism in a medium and culture conditions generally used for a genus or species of each microorganism, collecting them by the medium or by centrifuging or the like and then mixing them. You can create it. For example, a medium containing 2.5 g of yeast extract, 5.0 g of peptone, and 1.0 g of glucose (pH 7.0) per liter can be used.
However, for Paenibacillus sp. KB-06,
More nutrient rich medium, eg yeast extract 1.0 g, peptone 2.0 g, meat extract 1.0 g, glucose 10.0 per liter
A medium containing g (pH 7.4) is particularly preferable. Each of the microorganisms is preferably in the logarithmic growth phase in order to control the number of cells and shorten the time when mixing, but those in the stationary phase can also be used. The number of cells of each microbial species can be easily adjusted by preparing a growth curve in advance. In some cases, the microbial composition may be prepared without preparing it in a large amount at a time, and if necessary, a microbial composition (seed culture) to be used as a "seed" for large-scale preparation may be prepared in advance. For use in treating organic waste, the thus-prepared seed culture containing each microorganism can be mixed and further cultured to prepare a large amount of a mixture of microorganisms sufficient for treating organic waste. By predefining the medium and culture conditions for preparing a large-scale preparation from such a seed culture, it is possible to maintain a constant relationship between the seed culture and the composition of the large-scale preparation prepared therefrom. it can. For example, for any microbial composition containing the above-mentioned microorganisms (1) to (5) and (1) to (6), a general-purpose microbial culture medium, for example, yeast per liter at 35 ° C to 37 ° C. Extract 2.5g, peptone 5.0g, glucose 1.0g (pH 7.
It can be cultured using a medium containing 0). When cultured in such a medium, at least the above (1)-
The microorganisms of (5) have the same growth rate. If necessary, the composition of the bulk preparation from the seed culture can be ascertained by individually measuring the cell number or concentration of each microorganism under appropriate media and conditions.

Alternatively, each of the grown strains is collected by centrifugation or the like, suspended in physiological saline or an appropriate buffer, and then adsorbed on an appropriate adsorbent such as sterile rice bran or biodegradable porous material. Then, the microbial composition obtained by mixing them may be used as a seed culture (so-called seed culture) for large-scale preparation. In particular, biodegradable materials, for example, biodegradable porous balls such as "Bioball" available from Matsushita Electric Works, are excellent adsorbents, are easy to handle and have good shelf life, but are almost completely natural. It is particularly preferable because it decomposes. The microbial composition of the present invention may be adsorbed on such a biodegradable porous material. The composition may comprise a biodegradable porous material having each microbial species in the composition of the invention adsorbed separately,
The composition of the present invention may include a biodegradable porous material adsorbed with one or more microbial species. The original bacterium obtained by culturing such an elementary culture may be used as it is, but may also be dried at 40 ° C to 50 ° C for about 2 days to form a powder. The powdered product can be stored and is convenient for handling.

Medium for culturing these microorganisms, culture conditions,
Techniques for measuring the number of cells and the mass of cells are well known to those skilled in the art. Further, in the present invention, unless otherwise specified, the composition ratios are based on the mass ratio.
If necessary, the relationship between the number of bacterial cells and the mass of each bacterial cell can be measured in advance. Generally, the microbial composition of the present invention is adjusted so that the total number of microorganisms is about 1 × 10 ^{8 to} 1 × 10 ⁹ per 1 g based on the composition. The optimum condition corresponding to the content and state of the organic waste can be determined by using the microorganism of the present invention having a changed composition and conducting a decomposition test of the organic waste under the conditions according to the conditions described later. When always treating organic wastes of a certain content and condition, for example, when treating wastes from a particular food processing industry constantly, it is preferable to predetermine the optimum conditions by such a method. .
It is needless to say that this composition is a value at the start of the treatment and may change with time after the start of the treatment according to the treatment conditions and the content of the target degradation product.

The target of the decomposition method of the present invention is organic wastes, in particular so-called food waste including vegetable waste and the like.
It is preferably a biological waste mainly containing animal and plant residues including manure, vegetable scraps, fish roe, etc. in the livestock industry or food processing industry. Here, "organic waste" means waste mainly containing organic compounds, and "biological waste" means, as in the above-mentioned examples, organisms such as plants and animals, microorganisms, parts of organisms, It refers to excrement or organic waste mainly containing decomposed products derived from living organisms. The present invention can be used in general for decomposing organic wastes, particularly biological wastes, as long as it does not contain substances that are particularly harmful to microorganisms. The method for decomposing the microbial composition and the biological waste of the present invention is, for example, vegetable scraps, fruit peels, fish roe, food residues, okara, sake and shochu lees, livestock or small animal excrement, dead chickens, laboratory animals. It can be used for decomposition of animal carcasses, seafood, etc. and mixtures thereof.

The method for decomposing organic waste according to the present invention is characterized in that the aforementioned microorganisms are mixed with the organic waste and maintained at a constant temperature for a constant time. In the method of the present invention, first, the organic waste to be decomposed and the microbial composition of the present invention are mixed to adjust the water content, and generally, auxiliary materials are further added. This ancillary material also functions as an additional nutrient source for microorganisms and as a regulator of the water content of the organic waste to be treated. The mixing ratio of the organic waste and the microbial composition of the present invention is such that the total amount of microorganisms is about 0.3% by mass to about 10% with respect to the organic waste.
%, More preferably about 2% to about 8% by weight, and particularly preferably about 3% to about 7% by weight.
So that

In the method of the present invention, the water content at the start of waste treatment may range from about 40% to about 70%, but from about 40% to about 70%.
60% is preferable and about 40% to about 50% is particularly preferable. However, the higher the water content, the longer the drying step described later, so it is preferable to adjust the fuel consumption necessary for drying at the same time. The water content may be adjusted, for example, by simply adding water to the object to be treated, and the water quality is not particularly limited as long as it does not significantly inhibit the growth of microorganisms. Also,
When the water content of the object to be treated is high, the water content may be adjusted by adding an auxiliary material. In the method of the present invention, since the nutrients necessary for the growth of microorganisms may be insufficient only with the organic waste to be treated, it is preferable to add various auxiliary materials for supplementing the nutrients. Specifically, rice bran, defatted rice bran, okara, especially dried okara, soybean meal, defatted soybean meal, etc. can be used, and rice bran, defatted rice bran and soybean meal, and defatted soybean meal are particularly preferable. The amount of auxiliary materials added is approximately 20 with respect to the processed material
% To about 80% by weight is preferred, more preferably about 30
Wt% to about 70 wt%, particularly preferably about 40 wt% to about 60 wt%
It is% by mass. Generally, the higher the water content of the waste to be treated, the larger the amount of the auxiliary material, and the lower the water content of the waste to be treated, the smaller the amount of the auxiliary material.

The resulting mixture is then transferred to a warmable container, generally about 40 ° C to about 65 ° C on average, preferably on average.
It is heated to 45 ° C to 65 ° C, particularly preferably to an average of about 50 ° C to about 65 ° C. The above-mentioned temperature range is not absolute, and the temperature range preferable for the microbial composition of the present invention to function while suppressing the generation of other harmful microorganisms and particularly the temperature range suitable for the method of the present invention are exemplified. It is a thing. As long as the microorganism used in the present invention has a physiological activity, for example, since the microorganism in the microbial composition of the present invention is a thermotolerant bacterium, the microorganism of the present invention is completely killed temporarily during the decomposition treatment. The temperature may rise to a temperature that does not occur, for example, the temperature may rise to about 75 ° C for about 15 minutes, and conversely, the temperature may temporarily rise to 40 ° C during the decomposition treatment. May fall below. The container used in the method of the present invention preferably has at least a heating device, but more preferably has a stirring device.
The system and structure of this heating device are not particularly limited,
Warming with steam is preferred. In the case of heating with steam, it usually takes about 30 minutes to about 60 minutes to reach a predetermined temperature, and generally the temperature rising time in this range is preferable, but it can be changed if necessary. An apparatus suitable for such treatment is commercially available, for example, Kiyomoto Bio Co., Ltd. has commercially available apparatuses of various capacities as KB series. If one of such devices is used, it is possible to treat biological waste of about 4000 kg / day (specific gravity of about 0.6, water content of 65%). On the other hand, the present invention, by using a smaller device, for example about 400 kg / day discharged from a supermarket or the like, or about 50 ~ 100 kg / day biological waste discharged from general households or small stores It is also suitable for processing.

When the temperature reaches a predetermined temperature, it may be left as it is, but it is preferable to stir it. When stirring, continuous stirring may be performed, or stationary and stirring may be repeated at regular intervals. The time of standing or standing / stirring depends on the amount of organic waste to be decomposed and the amount of microbial composition to be used, but decomposes at a microbial composition / organic waste ratio within the above range. In general, about 1 to about 24
Time, preferably about 5 to about 20 hours, more preferably about 5
~ About 15 hours. Under the above-mentioned conditions, the decomposition treatment is usually completed within about 24 hours, and no further treatment is necessary, but treatment may be performed for more than 24 hours. The reason why the processing is not performed for a long time is mainly because the cost is suppressed. Therefore, the processing may be performed for a long time depending on the demand of the operating time zone of the device.

The stirring is performed in order to mix the microorganisms in the microbial composition of the present invention with the target organic waste well and to supply oxygen to the aerobic microorganisms.
Therefore, the stirring speed, stirring time and standing time can be arbitrarily selected and can be adjusted in consideration of the amount of organic waste, the shape of the container and the like. For example, it may be determined according to the conditions that can be used when culturing each microorganism in the present composition. Specifically, as standard conditions, for example, about 20 minutes to about 60 minutes, preferably about 30 minutes to about 60 minutes of standing and about 1 minute to about 5 minutes, preferably about 1 minute to about 3 minutes. The stirring times of can be combined. Further, it is preferable to additionally perform aeration during stirring.

After the lapse of the aforementioned period, a step of drying the decomposition product may be added. This drying process is about 80 ℃ ~ 100 ℃,
Preferably, the decomposition product containing the remaining microorganisms is heated at about 85 ° C to about 100 ° C, and the water content is about 20% or less, preferably about 20%.
It is a step of lowering to 15% or less, particularly preferably to about 12% or less. The heating and drying time can be adjusted as appropriate,
The drying process can be completed in about 2 to about 5 hours using an appropriate apparatus. By carrying out such heat treatment, harmful bacteria which may have grown during the decomposition treatment and plant seeds which may be an obstacle to reuse can be killed. Further, following the decomposition treatment of the present invention, foreign matters or residues not decomposed by the method of the present invention may be removed by sieving. For example, since bones, plant seeds, etc. may remain without being completely decomposed, it is preferable to separate and crush them. The crushing treatment can be performed before the decomposition treatment according to the present invention, but it is preferably performed after the decomposition treatment because the cleaning treatment inside the crusher is simple and the like. Biological waste such as crushed bone may be further decomposed by the method of the present invention or together with other organic waste.

After the drying treatment, the treated product may be cooled, and may be used or discarded as it is or after further processing depending on the use of the decomposition product. For example, it can be subjected to general waste treatment steps such as incineration and landfill, but the decomposition product obtained by the method of the present invention is used as it is, or after being processed and molded as necessary, fertilizer, soil conditioner, It can also be used as feed. For example, when it is used as a fertilizer or a soil conditioner, it can be used in the same manner as an ordinary compost or a soil conditioner. Depending on the contents to be processed and the disassembly processing conditions,
According to one embodiment of the present invention, about 400 kg of garbage discharged per day from a supermarket or the like can be used as a fertilizer or a soil conditioner by the microbial composition and the treatment method of the present invention. Degradation products are produced. Moreover, at least a part of the microorganisms contained in the composition of the present invention is still alive in the decomposition product of the method of the present invention, and can be used for compost fermentation of livestock excrement and the like. In particular, Paenibacillus sp. KB-06 is a heat-resistant bacterium and at the same time a nitrogen-fixing bacterium, which not only contributes to the decomposition of organic waste but also can fix nitrogen during and after the decomposition process. When using as a feed or fertilizer, it is particularly preferable to use a microbial composition containing Paenibacillus sp. KB-06 for decomposition treatment.
When carrying out such secondary use, 100
It is particularly preferable to heat to a temperature of not less than 0 ° C. to kill harmful microorganisms such as Escherichia coli and Salmonella before use.
Since the microbial composition of the present invention contains a thermotolerant bacterium as a useful microorganism, and at least a part of it remains in the decomposition product, a considerable number of useful microorganisms survive even when subjected to such a high temperature treatment.

[0024]

EXAMPLES Example 1. Production of microbial composition (1) Production of bio-ball bacterium 10 μl of cryopreserved strain (including 750 μl of 80% glycerol aqueous solution and 250 μl of bacterial cell culture solution) in 100 ml liquid medium (yeast extract 2.5 g /
l, peptone 5.0 g / l, glucose 1.0 g / l, pH 7.0). After culturing with shaking at 150 rpm at 45 ° C. for 48 hours, the cells were collected by centrifugation (4000 rpm, 30 minutes). The amount of cells obtained is as follows.

[Table 4] Table 3.

Each bacterial cell was suspended in 10 ml of a 0.89% NaCl aqueous solution in a 100 ml Erlenmeyer flask, and 5 g of Bioball (Matsushita Electric Works, about 0.4 mg / cell) was added until the water content disappeared with a microspatel. Stir well for about 5 minutes.
By this operation, about the same number of microorganisms are adsorbed per bio-ball for each microorganism species. The thus-prepared microbial-adsorbed bio-ball was called "bio-ball bacterium".

(2) Production of original bacterium for treating organic matter 25 g of bioball prepared in (1) (5 for each microorganism)
g), rice bran 5.5 kg, molasses 50-60 g, botanic (mineral water), and distilled water 3 l were put into a small breeder, and cultured at 35 to 37 ° C. for about 6 hours. The culture was dried at 45 ° C. for 2 days to give a powder. The water content of the obtained powder was about 10%. The thus-prepared microbial composition is called "original fungus". This original bacterium is 1.5x as a whole per 1g
It contained 10 ¹⁰ microorganisms.

Example 2. Treatment of fish ara The ara composition of the fish was treated with the microbial composition (former bacterium) obtained in the same manner as in Example 1. Ara 100.0 kg (water content 65.0%) of fish including tuna head, rice bran 50.0 kg (water content 12.
0%) and 5 kg of original bacteria are mixed to give a total amount of 155.0 kg (water content 46.1
%) And put into a 300 liter steam type fermenter (Kiyomoto Bio) of Green Gaia. The treatment process is as follows.

[Table 5] Table 4. Temperature and time in each process

In this step, the head of the fish was almost decomposed after about 4.5 hours, and a part of the bone remained.
After the drying process, most of the bone, including the remaining bone, was decomposed and 83.0 kg of a yellowish brown powdery product (water content 4
%) Obtained. The decomposition products had almost no odor.

Example 3. Nitrogen Compound Composition of Degradation Products The nitrogen compounds in the degradation products obtained in Example 2 were analyzed and the results are shown in FIG. As can be seen from FIG. 1, the free amino acid content was increased in the degradation products, while the ammonia and amine in the degradation products, which could cause malodor, remained almost unchanged before and after the treatment.

Example 4. Fertilizer and soil improvement of decomposition products
Use as an agent In the same manner as in Example 1, Paenibacillus sp. KB-06
2t of a microbial composition containing (FERM P-18368) was prepared. 4 t of fish roe, 2 t of this microbial composition, and 2 t of soybean meal were mixed, heated to about 60 ° C. using a steam boiler, and then treated for about 17 hours. Decomposition products at about 90 ℃ -100 ℃ 2
Dry for ~ 3 hours. The decomposed product after drying was cooled and used as a soil improver and a fertilizer according to a usual method. As a result, the damage of root-knot nematodes was suppressed in cucumber, and the sugar content was increased in the potted mango (up to approx.
20 degrees) was seen.

Reference Example 1. Bacillus licheniformis KB
Of Proteolytic Enzyme Secreted by -04 (1) Collagenase Activity Measurement (i) Search for Collagenase-Producing Bacteria A collagenase-producing bacterium was searched using an agar medium containing collagen as a sole carbon source and nitrogen source. Each strain was plated on the following agar medium and cultured at 30 ° C for 5 days, and the presence or absence of halo was used as an index of collagen-producing bacteria. The medium used is shown in Table 5. The results are shown in Table 6. In Table 6, the relative size of the halo is expressed as the number of "+".

[Table 6] Table 5. Agar medium for searching collagenase-producing bacteria

[0032]

[Table 7] Table 6. Halo formation

(Ii) Measurement of collagenase activity in collagenase-producing medium Collagenase-producing medium, that is, wheat germ 0.2
% -Collagen 1.0%, chitin 1.0%, pH 7.0, the culture supernatant was prepared by shaking culture of KB-04 at 30 ° C. for 5 days. 0.2 ml of the culture supernatant and 0.8 ml of 50 mM Tris buffer (pH 7.0) were added to 5 ml of the substrate (1% collagen, 4 mM CaCl2, 50 mM Tris buffer pH 7.0), and the mixture was reacted at 30 ° C for 2 hours. The reaction solution was filtered, and 0.5 ml of 0.2M citrate buffer (pH 5.0) and 1.2 ml of ninhydrin reagent ^* 1.2 ml were added to 1 ml of the filtrate. The mixture was boiled for 15 minutes and cooled. Further, 5 ml of 60% ethanol was added, and the absorbance (OD ₅₇₀ ) at ₅₇₀ nm was measured. As a negative control, an "enzyme blank" containing an enzyme but no substrate was prepared, and as a comparative control, a "substrate blank" containing a substrate but not adding an enzyme was prepared. The measurement results are shown in Table 7. * Ninhydrin reagent A solution: 0.01M KCM solution 5ml Add methylcellosolve to make the total volume 250ml Solution B: Dissolve 2.5g ninhydrin in 50ml methylcellosolve Solution A: Solution B immediately before use Mix at a ratio of 5: 1.

[0034]

[Table 8] Table 7 Collagenase activity of Bacillus licheniformis KB-04 culture supernatant

(2) Measurement of protease activity Wheat germ 0.2%, collagen 1.0%, chitin 1.0% (pH 7.
After shaking culture at 150 rpm for 5 days at 30 ° C. in a liquid medium containing 0), centrifugation (4000 rpm, 30 minutes) was performed to prepare a culture supernatant. The protease activity was measured using this culture supernatant. 0.9 ml of 50 mM Tris buffer (pH 7.0) was added to 0.1 ml of the culture supernatant, and preheated at 30 ° C. for 5 minutes. Substrate (casein 0.
5% of 6%, 0.05M Na ₂ HPO ₄ , pH 7.0) was added and reacted at 30 ° C. for 10 minutes. Add 5 ml of 0.44M trichloroacetic acid and add 30
It was allowed to stand at 30 ° C for minutes. Next, the reaction solution was filtered, and the absorbance (OD ₂₇₅ ) of the obtained filtrate at 275 nm was measured. The results are shown in Table 8.

[0036]

[Table 9] Table 8. Protease activity of Bacillus licheniformis KB-04 culture supernatant

From these results, it is clear that the culture supernatant of KB-04 has strong protease activity and collagenase activity.

(3) Pork and pig skin disintegration experiment Using the crude enzyme solution obtained in a collagen liquid medium (1)
A pork / pork skin disintegration experiment was carried out in the same manner as in. Commercially available pork and pork skin were cut into 1 cm square pieces, and both pork and pork skin were boiled. Put pork and pig skin in each test tube, add 5 ml of the supernatant (crude enzyme solution) obtained by liquid culture, add a few drops of toluene on it, cover with a rubber stopper, and react at 45 ° C for 24 hours Then, the degree of disintegration was visually observed. As a control, the one to which the culture supernatant was not added was used. The results are shown in Table 9 below.

[Table 10] Table 9. Degradation of pork and pig skin The use of collagen liquid medium also increased the degree of pork disintegration.

Reference Example 2. Bacillus subtilis KB-01
Analysis of secreted proteolytic enzyme (1) Collagenase activity measurement Bacillus subtilis KB-01 is a medium for collagenase production, collagen 1.0%, chitin 1.0%, HEART INFUSI
Liquid medium containing ON powder 0.2% (pH 7.0) at 30 ℃ for 5 days,
After shaking culture at 150 rpm, centrifugation (4000 rp
m, 30 minutes) to prepare a culture supernatant. Using this culture supernatant, Reference Example 1. The collagen activity was measured in the same manner as in (1). The results are shown in Table 10.

[Table 11] Table 10. Collagenase activity of culture supernatant of Bacillus subtilis KB-01

(2) Protease activity A culture supernatant was prepared in the same manner as in (1), and Reference Example 1.
Protease activity was measured in the same manner as in (2). The results are shown in Table 11.

[Table 12] Table 11. Protease activity of culture supernatant of Bacillus subtilis KB-01

(3) Pork / pork skin disintegration experiment A culture supernatant was prepared in the same manner as in (1), and Reference Example 1.
A pork / pork skin disintegration experiment was conducted in the same manner as in (3). No culture supernatant was added to the control. The results are shown in Table 12
Shown in.

[Table 13] Table 12. Degree of pork and pig skin collapse From these results, it is clear that the culture supernatant of KB-01 has strong protease activity and collagenase activity.

[0042]

EFFECTS OF THE INVENTION By using the microbial composition and the method for decomposing organic waste of the present invention, it is possible to decompose an organic waste, particularly a biological waste, in an extremely short time, and to eliminate a bad odor during the decomposition. Occurrence can be suppressed. further,
The decomposition products obtained are fertilizers, soil conditioners, fish feed,
It can also be used as an inoculum for compost fermentation of livestock excrement and the like, and these can be provided at low cost.

[Brief description of drawings]

FIG. 1 is a composition of nitrogen compounds containing amino acids in a decomposed product. Amino acids are shown in three letters according to the usual notation. P-Ser: phosphoserine, Tau: taurine, PEA: phosphoethanolamine, Sar: sarcosine, a-AAA: α-aminoadipic acid, Cysthi: cystathionine, b-Ala: β-alanine, b-AiBA: 3-aminoisobutyric acid , G-ABA: γ-aminobutyric acid, EOHNH2: ethanolamine, Orn: ornithine, 3
Mehis: 3-Methylhistidine, Car: Carnosine, Hypr
o: hydroxyproline

Front page continuation (51) Int.Cl. ⁷ Identification code FI theme code (reference) // (C12N 1/20 C12R 1:10 C12R 1:07) 1: 125 (C12N 1/20 C12R 1:01 C12R 1 : 10) B09B 3/00 ZABD (C12N 1/20 C12R 1: 125) (C12N 1/20 C12R 1:01) F term (reference) 4B065 AA15X AA19X AC01 AC14 AC20 BB22 BC42 CA55 4D004 AA03 AA04 BA04 CA18 CA22 CC07 CC08 DA02 DA03 DA06 DA09 4H061 AA02 CC47 CC55 EE61 EE66 FF06 GG18 GG41 GG48 LL02

Claims (16)

[Claims] 1. A microbial composition containing the following microorganisms (1) to (5): (1) Bacillus subtilis (2) Bacillus sp. KB-02 (FERM P-18364) (3) Bacillus varismorti Sus (4) Bacillus licheniformis (5) Bacillus pumilus 2. (1) The accession number is Bacillus subtilis
FERM P-18363, (3) Bacillus varismortis identified by accession number FERM P-18365, (4) Bacillus licheniformis identified by accession number FERM P-18366, (5) Bacillus pumilus Is the consignment number FERM P-183
The microbial composition according to claim 1, which is a microorganism specified by 67. 3. Further, (6) Paenibacillus sp. KB-06
The microbial composition according to claim 1 or 2, which comprises (FERM P-18368). 4. The method according to claim 1, wherein the microorganisms are contained in substantially equal amounts.
The microbial composition according to any one of 1. 5. The microbial composition according to claim 1, wherein the microorganism contained in the composition is adsorbed on the biodegradable porous material. 6. The microbial composition according to claim 5, wherein each of the microorganisms (1) to (6) is adsorbed on a separate biodegradable porous material. 7. The microorganism composition according to claim 5, wherein two or more kinds of microorganisms among the microorganisms of (1) to (6) are adsorbed on the same biodegradable porous material. 8. Mixing (i) an organic waste, an additional nutrient source for microorganisms and a water content regulator of said organic waste, and a microbial composition according to any one of claims 1-7. (Ii) heating the temperature of the mixture obtained in (i) to 40 ° C to 65 ° C (iii) maintaining an average temperature of 40 ° C to 65 ° C. Method of decomposing organic waste. 9. Mixing (i) an organic waste, an additional nutrient source for microorganisms and a water content regulator of said organic waste, and a microbial composition according to any one of claims 1-7. Step (ii) heating the temperature of the mixture obtained in (i) to 40 ° C to 65 ° C, (iii) stirring continuously or at regular intervals while maintaining the average temperature of 40 ° C to 65 ° C. A method for decomposing organic waste, comprising: 10. The method according to claim 8, wherein step (i) is carried out at a water content of 40% by mass to 65% by mass. 11. The method according to claim 8, wherein the additional nutrient source for microorganisms and the water content regulator of the organic waste are selected from rice bran, defatted rice bran, okara, dried okara, soybean meal and defatted soybean meal. The method described. 12. The method according to any one of claims 8 to 11, wherein the organic waste is biological waste. 13. Decomposition product of organic waste produced by the method according to any one of claims 8 to 12. 14. Use of a microbial composition containing the following microorganisms (1) to (5) for decomposing organic waste. (1) Bacillus subtilis (2) Bacillus sp. KB-02 (FERM P-18364) (3) Bacillus varismortis (4) Bacillus licheniformis (5) Bacillus pumilus 15. (1) Bacillus subtilis is identified by accession number FERM P-18363, (3) Bacillus varismortis is identified by accession number FERM P-18365, and (4)
Bacillus licheniformis was identified by accession number FERM P-18366 and (5) Bacillus pumilus was assigned accession number FERM P
The use according to claim 14, which is a microorganism identified by -18367. 16. (6) Paenibacillus sp. KB-
Use according to claim 14 or 15, using 06 (FERM P-18368).