KR20190028407A - Composition for decomposing a food waste - Google Patents

Abstract

The present invention relates to microorganisms for decomposing and eliminating food waste, a composition for decomposition and elimination comprising the same, and an eliminating method of food waste using the same, wherein the food waste is completely decomposed by the microorganisms, thereby reducing odors as much as possible. The composition of the present invention has activities of amylase, cellulase, protease, and lipase; and comprises Bacillus subtilis DYG-B11, Bacillus subtilis DYG-B12, Lactobacillus sp. DYG-B13, and Bacillus subtilis DYG-B14, wherein the microorganisms have excellent food waste decomposing activities.

Description

{Composition for decomposing food waste}

The present invention relates to a food waste disintegration microorganism, a composition for decomposing and dissolving the microorganism, and a method for disposing of food waste using the composition. More particularly, the present invention relates to a food waste disintegration microorganism which is resistant to acid, , Protein, and cellulose. It is a microorganism suitable for the decomposition and destruction of food garbage. It is a microorganism that decomposes and completely destroys food through fermentation of food garbage, And a method for dissolving food waste using the composition.

Food waste is generated in large quantities due to rapid population growth and improvement of living standards. At present, the incidence of food waste in Korea is about 20% or more of total municipal waste, of which more than 60% Garbage is being discharged from the household, and the issue of food waste disposal has long been a serious social problem. Especially, in large cities where disposal facilities or disposal places or methods are inadequate, the issue of food garbage disposal is getting serious. to be. On the other hand, a method of recycling food waste to feed or compost, etc. has been widely used in recent years as a solution for food waste which has been used conventionally. However, it is difficult to use food waste due to generation of odor, The landfill and incineration still used in most food waste disposal methods have various problems due to the depletion and incineration of the landfill site, that is, the incinerator temperature is lowered due to moisture contained in the food waste during general incineration, and dioxin and dibenzofuran There is a problem that harmful gas is generated.

In this situation, it has been suggested that the cost of several trillion won (KRW) will be required to process food waste in Korea in the future. Methods for reducing household garbage have been proposed. Fermentation and extinction methods. The dry compression method is a method in which water, which occupies 75% of food waste, is dried by heating and then physically compressed to reduce its volume. However, since the organic matter remains, there is a problem that environmental pollution is caused again after landfilling. The extinction method is a method in which organic materials in waste are decomposed into water and carbon dioxide by using agitation and decomposition ability of microorganisms. Recently, interest has been increasing and it has been proposed to use various microbial agents. For example, In Publication No. 2002-0017873, there is disclosed a process for producing a food waste, which comprises the steps of: depositing Bacillus amyloliquefaciens, Bacillus subtilis, Bacillus subtilis, and Saccharomyces cerevisiae, which are deposited under accession number KFCC-11204 &Quot; 1: 1: 1: 1 mixed microbial fermentation medium MDG200 and their culture method " Korean Patent Laid-Open Publication No. 2012-0041377 discloses "a microorganism preparation for eliminating fermentation treatment of food waste under aerobic conditions, comprising Leuconostoc mesenteroides and Lactobacillus delbrueckus sp. (Lactobacillus delbrueckii subspecies bulgaricus), "which discloses a microbial preparation for aerobic elimination fermentation treatment of food waste.

The use of a microorganism having an excellent decomposing activity of organic matter as a disinfection accelerator made of a microorganism preparation having various compositions as described above can solve a number of problems in the treatment of food garbage. However, The present inventors have found that a large amount of fermentation mass remains in the state that the fermentation decomposition of the food waste is completed, Thereby still causing problems due to the processing of these masses. In addition, there is still a problem of odor generation in the treatment of household food waste.

Therefore, the inventors of the present invention selected strains having excellent fibrinolytic, starch and protein decomposing properties, which are the main constituents of food waste, for optimal treatment of food waste, There is provided a food waste disassembling microorganism comprising a new microorganism preparation capable of maximizing efficiency and capable of exerting an excellent role in environment improvement such as eliminating harmful substances and removing odor, and a method for providing a composition for decomposing and dissolving it As a result, the present invention has been completed.

Patent Document 1: Korean Patent Publication No. 2002-0017873 Patent Document 2: Korean Patent Laid-Open Publication No. 2012-0041377

SUMMARY OF THE INVENTION Accordingly, the present invention has been made in view of the above problems of the prior art, and it is an object of the present invention to provide a food waste which is resistant to acid, alkali and salt, The present invention relates to a microorganism suitable for digestion and destruction of food waste having a strong decomposition activity and capable of decomposing food waste and decomposing and completely extinguishing food through fermentation of food waste, will be.

Another object of the present invention is to provide a composition for disassembling and dissolving food waste comprising a group of microorganisms having the excellent food waste decomposition and extinguishing ability and a component capable of removing harmful substances and removing odors.

It is another object of the present invention to provide a method for extinguishing food waste using a composition for decomposing and dissolving food waste comprising a microorganism group having the excellent food waste decomposing ability.

The present invention may also be directed to accomplishing other objects that can be easily derived by those skilled in the art from the overall description of the present specification, other than the above-described and obvious objects.

In order to accomplish the above object, the present invention provides a food waste disassembling microorganism;

Amylase, cellulase, protease and lipase enzyme having a degrading activity of the active food waste is excellent Bacillus subtilis DYG-B11 (Bacillus subtilis DYG- B11) strain: and characterized in that (Accession No. KCTC 11085BP).

In order to accomplish the above object, the present invention provides a food waste disassembling microorganism;

Amylase, cellulase, protease and lipase enzyme having a degrading activity of the active food waste is excellent Bacillus subtilis DYG-B12 (Bacillus subtilis DYG- B12) strain: and characterized in that (Accession No. KCTC 11086BP).

In order to accomplish the above object, the present invention provides a food waste disassembling microorganism;

Amylase, cellulase, protease and lipase enzyme having a degrading activity of the active food waste is excellent Bacillus subtilis DYG-B14 (Bacillus subtilis DYG- B14) strain: and characterized in that (Accession No. KCTC 11088BP).

In order to accomplish the above object, the present invention provides a food waste disassembling microorganism;

Amylase, cellulase, protease and lipase enzyme having a degrading activity of the active food waste is excellent Lactobacillus sp DYG-B13 (Lactobacillus sp DYG- B13.) Strain: and characterized in that (Accession No. KCTC 11087BP).

According to another aspect of the present invention, there is provided a composition for decomposing and dissolving a food waste decomposition microorganism comprising;

Amylase, cellulase, protease and has a lipase activity food waste decomposing activity is excellent Bacillus subtilis DYG-B11 (Bacillus subtilis DYG-B11) strain (Accession No: KCTC 11085BP), Bacillus subtilis DYG-B12 (Bacillus subtilis DYG-B12) strain (Accession No: KCTC 11086BP), Lactobacillus sp DYG-B13 (Lactobacillus sp DYG-B13) strain (Accession number:. KCTC 11087BP) and Bacillus subtilis DYG-B14 (Bacillus subtilis DYG-B14) Strain (Accession No .: KCTC 11088BP).

According to another aspect of the invention, the composition is characterized in that the bulk holde less Sepharose when Ah (Burkholderia cepacia), further comprising a three Levy MY process jiae (Saccharomyces cerevisiae) a saccharide.

According to another aspect of the present invention, there is provided a method for disposing food waste using a composition for decomposing and dissolving a food waste decomposing microorganism, comprising:

Amylase, cellulase, protease and has a lipase activity food waste decomposing activity is excellent Bacillus subtilis DYG-B11 (Bacillus subtilis DYG-B11) strain (Accession No: KCTC 11085BP), Bacillus subtilis DYG-B12 (Bacillus subtilis DYG-B12) strain (Accession No: KCTC 11086BP), Lactobacillus sp DYG-B13 (Lactobacillus sp DYG-B13) strain (Accession number:. KCTC 11087BP) and Bacillus subtilis DYG-B14 (Bacillus subtilis DYG-B14) And a strain (Accession No .: KCTC 11088BP) is used to decompose and destroy food waste.

The method of the present invention for decomposing and dissolving food waste, the composition for decomposing and dissolving the food waste, and the method for dissolving food waste using the same, is characterized in that the composition is made of starch, fat, protein, The present invention provides a microorganism group suitable for decomposing and extinguishing food garbage having a strong decomposing activity evenly over all of cellulose, and a composition for decomposing and dissolving garbage by adding a component capable of removing harmful substances to these microorganisms The food waste is completely decomposed by the microorganisms so that the odor can be minimized while the food garbage is decomposed and completely destroyed so that the remaining amount of the food waste can be treated so as to remain almost unchanged.

Figure 1 represents the results of the four kinds of microorganism selected and deposited according to the present invention, Figure 1a is a Bacillus subtilis DYG-B11 is for the (Bacillus subtilis DYG-B11), Figure 1b is a Bacillus subtilis DYG -B12 is for the (Bacillus subtilis DYG-B12), Figure 1c is a Lactobacillus sp DYG-B13 (Lactobacillus sp. DYG -B13) are for a, Figure 1d is a Bacillus subtilis DYG-B14 (Bacillus subtilis DYG- B14 ), ≪ / RTI >
FIG. 2 is a graph showing the decomposition ratio of food waste using the composition for decomposing and dissolving food waste according to a preferred embodiment of the present invention, and shows a remaining mass after decomposition and disappearance with time.

Hereinafter, the present invention will be described in more detail with reference to preferred embodiments with reference to the accompanying drawings, but it goes without saying that the scope of the present invention is not limited thereto.

To solve the problems of the prior art described above, in order to select a strain having excellent decomposing ability of food waste according to the present invention, a sample was collected from a composting plant for composting food waste as a main material, and a dominant microorganism was isolated in a laboratory . Each of the isolated microorganisms was tested for their resistance to salinity, acidity and temperature, enzyme activity test for starch, fat, protein and cellulose, and the extinction test for food waste. Excellent microorganisms were selected from compost.

In addition, the present invention has developed and provided a fungus for fermentation of food garbage, which is prepared by adding the nutrients for promoting microbial propagation to the microorganisms selected as described above at an appropriate concentration. The fungus according to the present invention is characterized in that the protease and lipase are positive, according to the characteristics of the food waste excellent activation of the cellulase and amylase, heat-resistance, salt tolerance, acid resistance and novel Bacillus subtilis DYG-B11, which has to my basic (Bacillus subtilis DYG-B11) strain, Bacillus subtilis DYG-B12 ( Bacillus subtilis DYG-B12) strain, Lactobacillus sp DYG-B13 (Lactobacillus sp. DYG -B13) strain and Bacillus subtilis DYG-B14 (Bacillus subtilis DYG- B14) accession number for each, Korea Research Institute of Bioscience and biotechnology as a strain KCTC 11085BP , KCTC 11086BP, KCTC 11087BP and KCTC 11088BP, and the fermentation of the food waste containing the strain The pragmatic destroy microbial agent is provided.

In the present invention, in particular, the composition of food waste in Korea is higher in salinity and acidity due to foods such as soybean paste, soy sauce, and kimchi compared with western food waste, and the content of cellulose is higher than that of meat, As described above, when considering the problem of deodorization in the fermentation extinction of food wastes and the characteristics of Korean food wastes as described above, it is important that the food wastes The fermentation destruction microorganism itself is important. It is based on the fact that the potency of amylase, cellulase, protease, lipase, etc., and the activity of protease, lipase and the like are particularly important for suppressing odor generation.

Among the four functional microorganisms selected according to the present invention, the novel KCTC 11085BP, KCTC 11086BP and KCTC 11088BP are gram-positive bacterium belonging to Bacillus subtilis, which is also called Bacillus subtilis. It is used for the fermentation of Chungkukjang, and many subspecies and micro variants are growing in various places. In addition, these microorganisms have been widely used as a variety of genetically modified microorganisms together with Escherichia coli, and have the ability to survive and adapt in various environments due to the characteristic of forming spores. In particular, the enzymes that degrade proteins are well developed in these bacteria, so the ability to ferment soybean protein was used in the production of chungkukjang. In addition, the species that develop enzymes that decompose various polymers according to subspecies are highly dominant microorganisms in places where various substances such as compost are mixed because they have the ability to effectively decompose fibrin and collagen. The origins of these donated microorganisms were also sampled at a food waste remediation plant. These microorganisms can be produced by subculturing in a culture medium containing a polymer, followed by primary separation and subculture for 3 to 4 times to pick out species having excellent polymer degradation ability and freeze-drying. These microorganisms are safe microorganisms of Bio Safety Lv.1 that can be dealt with without knowledge of microorganisms.

In addition, the strains KCTC 11087BP screening as described above is based Lactobacillus (Lactobacillus sp.) With an anaerobic, gram-positive bacilli are all capable of growth under aerobic conditions. These microorganisms do not form spores but are microorganisms capable of survival for a long time if the temperature conditions are met. This is due to the characteristic of producing weak acids and making the environmental conditions favorable to them. The KCTC 11087BP strain is a microorganism which preferably occupies about 3 to 5% in the composition for digesting food poisoning according to the present invention and is characterized in that the substance which is difficult to decompose Bacillus strain is made to be easily decomposed through weak acid production have. The origin of these bacteria is easy to think of as yogurt, but it is also a soil-derived microorganism such as Bacillus. It can survive in harsh environments unlike yogurt germs and is used as auxiliary bacteria in various fermentation processes. This strain is also a safe microorganism of Bio Safety Lv.1 that can be handled without knowledge of microorganisms.

According to the present invention, all four functional microorganisms selected in the above manner are collectively sampled in a composting plant where compost is made from food waste as the main material, and one gram of the sample is suspended in 10 ml of physiological saline for 3 to 4 hours The supernatant was suspended in 50 ml of Nutrient Broth (NB), and the mixture was stirred at 28 ° C., and the culture was diluted 10-fold to 4-fold to 10-fold to 7-fold The cells were cultured at 28 ° C in a nutrient agar (NA) containing 0.5% starch at a temperature of 28 ° C. Then, seven of the colonies were visually distinguished from each other, And the four kinds of starches having the best decomposition ability of starch were selected. The thus isolated seeds can be stored frozen at -70 ° C in a culture medium containing 50% glycerol and can be taken out and used for experiments.

As described above, the microorganisms selected according to the present invention are excellent in decomposing activity by high activity, have a wide range of survival temperature, survive at high salt concentration, exhibit the most active activity in the optimum pH range of 5.0 to 9.0 and exhibit the activation of protease and lipase And it was evaluated as a resistant strain of acid tolerance, acid resistance, alkali resistance and heat resistance, which is excellent in decomposition ability according to the characteristics of food waste in Korea due to production of cellulase and amylase.

According to another preferred embodiment of the present invention, the composition comprising microorganisms having excellent fermentation ability of food garbage according to the present invention can be used for various microorganisms having excellent protein, starch, fat and cellulose decomposing properties, that is, The present invention relates to a composition of a complex microorganism preparation prepared by mixing three types of Bacillus subtilis according to the present invention, one kind of Lactobacillus species, a bacterium, yeast and the like. Further, in order to exhibit a strong fermentation ability in a device for maintaining vitality in a distribution period and for dissolving food garbage, each microorganism is cultured in a liquid state, and then a solid (for example, zeolite, activated carbon, It is preferable to use a new manufacturing method in which the viable cell count and vitality are enhanced by selecting the lyophilization method unlike the conventional method of culturing.

Concrete using the strain of the composition according to the present invention is Bacillus subtilis (Bacillus subtilis) 3 type, bulk holde less Sepharose cyano (Burkholderia cepacia), three Levy in Saccharomyces My process jiae (Saccharomyces cerevisiae), Lactobacillus (Lactobaillus sp. ).

The microbial culture medium composition and culture conditions according to the present invention are as follows. That is, the microorganisms were inoculated with 1% of seedlings in the medium composition or a slightly modified medium according to the strains shown in Table 1 below, cultured at 30-37 ° C for 18-30 hours while being aerated and stirred and then cultured using a continuous centrifuge The cells are recovered.

Ingredients % Peptone 2 Yeast extract One Glucose 2 Sodium acetate 0.1 Ammonium citrate 0.1 Sodium carbonate 0.05 K2HPO4 0.1 MgSO4 0.01 MnSO4 0.005 ZnSO4 0.001

The recovered cells are mixed with an appropriate amount of a protecting agent (maltodextrin, trehalose, glucose, skim milk), rapidly frozen at -40 ° C or lower for about 24 hours, and then freeze-dried. The lyophilized microorganisms are homogeneously pulverized using a sieve of 100 μl. The above-mentioned freeze-dried microorganism seeds are mixed with an appropriate amount of a moisture regulator (eg, cedar chips) to maintain the number of microorganisms at 10 ^6-9 cfu / g Thereby preparing a food waste decomposing and dissolving composition.

Hereinafter, the processes for production of microorganisms and commercialization thereof according to the present invention will be described in detail.

1. Fermenter check

1-1 Check the cleanliness of the tank, the water used, the steam condition, and the air pressure before feeding the ingredient.

1-2 If the amount of steam or usage is bad, report to the production manager without putting the badge.

2. Fermenter tank cleaning and sanitization

2-1 Clean the inside of the tank (300L or 1,200L capacity) before feeding the medium.

2-2 Air sterilization refers to sterilizing the tank with only a few tens of thousands of tubes without the medium components.

2-3 sterilization temperature should be more than 15 minutes at 121 ℃ and 1.2 atm.

2-4 Air disinfection should be carried out when monitoring is necessary or if the culturing strains for each tank are changed.

CIP treatment of 2-5 tanks should be carried out in the same way as air disinfection.

3. Badge weighing

3-1 Weigh the contents so that the contamination with foreign substances can be completely suppressed.

3-2 Consideration should be given to incorporation of the culture well.

3-3 Weigh according to the amount of culture, weigh it in error, and report it to the production manager when it is added.

Add the media components to the 3-4 culture water in the order of peptone, enzyme extract and glucose, and dissolve at 60 ℃ until completely dissolved.

4. Medium sterilization and cooling

4-1 Microorganism Highly concentrated The most important step in pure culture is to be completely sterilized.

4-2 Sterilization of microorganisms should be at least 15 minutes at 121 ° C and at least 15 minutes. If sterilization is not carried out, the sterilization time should be prolonged and sterilized.

4-3 Make sure that no negative pressure is applied during cooling after sterilization.

4-4 After sterilization, a positive pressure of 0.5 vvm or more should be maintained so that no negative pressure is applied during cooling.

4-5 Amount of water used before sterilization of the medium and amount of water used after sterilization of medium should be the same.

5. Inoculation of microorganisms

5-1 It refers to the process of putting microorganism seeds into the tank.

It is assumed that 5-2 seeds have finished inspection of pollution.

5-3 Use a flame when inoculating the sterilized culture medium and release the pressure in the tank to prevent accidents caused by flame.

5-4 Use a clean flame glove when inoculating, and the inoculator does not cause factors affecting microorganisms such as chatter.

5-5 Check the culture conditions such as pH, temperature and rpm before inoculation. For example, in the case of Bacillus subtilis, the conditions of pH 7.0, 37 ° C and 120 rpm are fixed.

6. Culture

6-1 Cultivation refers to culturing a culture according to culture conditions (pH, temperature, rpm, etc.).

6-2 Aerobic strains should be cultured at optimal temperature (25-37 ℃) or optimum culture time (18-30 hours) for each strain between pH 6.0-7.0 while maintaining a positive pressure of 0.5 vvm or more.

6-3 Record changes in pH, temperature, rpm, etc. during incubation on the record.

7. Pollution Inspection

7-1 The contamination test is to identify the contamination of the final culture. Firstly, it is examined under a microscope and the presence of contamination is determined by pouring method through dilution of the culture.

7-2 It is not allowed to carry out the fermentation products that have not been inspected.

8. Centrifugation

8-1 When the culture is completed, the culture is centrifuged using a continuous centrifuge for the purpose of recovering the cells.

8-2 Centrifugal bowl is sterilized with steam to prevent contamination of other bacteria.

8-3 Before using the centrifuge, check the electric and motor conditions.

8-4 After inserting the ball, centrifuge about 100L per hour by high-speed rotation of 16,500rpm.

8-5 If an abnormality is detected during centrifugation, report it immediately to the production manager after action.

9. Protective Mixture

9-1 Protective agents are used to protect microorganisms under rapid freezing conditions of microorganisms.

9-2 Protective agents (maltodextrin, trehalose, glucose, skim milk) must be sterilized.

9-3 Care should be taken to prevent foreign matter from entering when mixing protective agent.

9-4 The protective agent mixing procedure is carried out until thoroughly mixed with the microorganism cells obtained above.

10. Freeze-drying and grinding

10-1 Frozen microbial cells mixed with the preservative are subjected to rapid freezing at -40 ° C or lower for 24 hours and the temperature of the deep freezer, which is a freezing device during freezing, is always checked.

10-2 Gloves should be used as there is a danger of frostbite when hand is frozen.

10-3 After 24 hours, the frozen cells are placed in a freeze dryer and lyophilized for 3-4 days. The lyophilization is performed at a shelf temperature of 30 ° C, a cold trap condenser of -70 ° C or lower, and a vacuum of 3-4 days while maintaining a vacuum of 15 mTorr or less.

10-4 Check the temperature and vacuum condition of the freeze dryer using the recording paper of the dryer.

10-5 After the dried microorganism is pulverized, it is homogeneously pulverized using a sieve of 100 μl.

10-6 Be careful not to mix the foreign materials in the crushing process.

10-7 The microbial origin is vacuum packed and stored at 4 ℃.

11. Moisture Control Mixing and Manufacturing

11-1 Increasing the fermentation speed and efficiency by increasing the contact area with food by using oak or cedar chip (size, 3-5mm) as moisture regulator to prepare composition for fermentation of microorganism fermentation.

11-2 In order to prepare the above-described composition, a scale was prepared according to a predetermined mixing ratio (10 ^6-7 cfu / g of microbial origin, 0.1% KH ₂ PO ₄ , 2% glucose and the remainder wood chips) To prepare the raw materials.

11-3 Mix in a mixer (screw mixer) and mix thoroughly for 30-60 minutes.

11-4 Weigh 5kg, 10kg, or 20kg each, put it in the wrapping paper and pack it.

11-5 Packaged products should be kept in a cool place so that they can be picked up before delivery.

As described above, the composition for fermentation annihilation containing the microorganisms manufactured according to the present invention is composed of a system for decomposing and discharging organic matter of food waste by controlling an appropriate moisture content under aerobic conditions.

The microorganisms of the composition according to the present invention are mainly composed of microorganisms of the Bacillus family. Such microorganisms are excellent in stability and able to adapt to various environmental changes such as surviving spores when the environment is poor. It is a microorganism that shows good adaptability even in strong foods and spices. Also, the microorganism of this line is a microorganism that has traditionally been used in fermented foods such as meju, and is not harmful to the general public.

The microorganism according to the present invention exhibits active activity of protease and lipase, and it is possible to fundamentally suppress the generation of malodor. In other words, the main cause of the malodor is incomplete fermentation decomposition in the decomposition process of protein and fat, and most of the odor is generated. Therefore, it can induce complete decomposition due to the active activity of protease and lipase, In addition, complete deodorization can be achieved with the deodorizing device in the extinction device.

In addition, the composition comprising the novel microorganisms according to the preferred embodiment of the present invention maintains the aerobic characteristics of the constituting microorganisms to the maximum to induce optimum decomposition and extinguishing by the organic interaction of the influencing factor and the decomposition media In order to prevent the cooling of the internal temperature, it is necessary to increase the efficiency of oxygen supply, to prevent microbial activity and decrease of air permeability, to maintain the proper moisture content in order to suppress the occurrence of anaerobic decomposition characteristics, / N ratio of 25 to 30: 1.

According to the present invention, the new microorganisms selected as having excellent fermentation extinction ability and deodorizing ability of food waste were classified into Bacillus subtilis and Lactobacillus species as described above, and their microbiological properties and nucleotide sequence It was identified as a novel strain.

Hereinafter, the present invention will be described in more detail with reference to examples and comparative examples, but it goes without saying that the scope of the present invention is not limited to these examples.

Example 1. Microbial screening

In order to select microorganisms which are excellent for fermentation of food waste, microorganisms collected from composting facilities were repeatedly cultivated several times to find 5 to 6 dominant species. These microorganisms are re-cultured in a dedicated degradation medium to select the species that exhibits the most ability to decompose organic matter. When the microorganisms are grown in a medium containing common organic matter decomposition and starch, fat, etc., the higher the decomposition ability, the more transparent decomposition of a wide range, and the desired microorganisms can be selected.

After the above-mentioned process, the microorganism which was proved to be an excellent microorganism was purified again and seeded.

Example 2. Microbial Preservation

The seeds isolated in Example 1 were largely managed in two ways. First, pure seeds were diluted in a liquid medium containing 50% of glycerol and divided into 1 ml. Twenty seedlings were stored in a refrigerated warehouse at -70 ° C. These prepared microorganisms are safely preserved for three years.

The seeds for the product are prepared by coagulating the PCA or Nutrient medium, coating the microorganisms thereon, culturing them at 30 ° C for 48 hours, storing them in a refrigerator for 2 to 3 months, and using them at any time.

When the microorganisms spiked on the PCA medium are consumed or after the preservation period has passed, the microorganisms stored at -70 ° C can be taken out and used again in about 30 pieces. When the number of microorganisms stored at -70 DEG C is reduced to 5, it is preferable to cultivate the microorganism again to maintain the microorganism.

Example 3: Production of microorganisms

Cultivate 5 liters in sterile condition. The cultured seeds are cultured in 100-liter units for 24 hours, and finally cultivated in a large-scale culture tank of 1 ton.

The produced microorganisms are remixed according to the amount of the product, and are passed through a large centrifuge to the freeze-drying device. The lyophilized microorganisms can be stored for 2 years or more in a non-humid environment, which helps to maintain the quality of the product.

In addition, the lyophilized microorganisms can be preserved in a sealed container until the product is manufactured, and can be easily used in the production of a large amount of products because the microorganism is dried after the compounding is completed.

Example 4:

1. To prepare the BHI (Brain Heart Infusion) agar medium shown in Table 2, 37 g / L of BHI medium was added to distilled water and mixed thoroughly. Add about 1.5% agar, shake occasionally with heat, and if necessary, heat completely to dissolve the media components. It is then sterilized at 121 ° C for 15 minutes.

(g / L) Calf brains 7.7  Beef heart 9.8  Proteose peptone 10  Dextrose 2  Sodium chloride 5  Disodium phosphate 2.5

2. After sterilization, when the medium temperature reaches 55 ° C, pour 20 ml of the medium into a plastic Petri dish in a clean bench, and allow to stand at room temperature for about 30 minutes. After the culture medium is completely hardened, the culture medium lid is turned upside down, and the culture medium is put in a 30 ° C incubator and dried for a day. Four new microorganisms according to the present invention are each plated on BHI (Difco) agar medium prepared in the above process, and cultured at 30 ° C for about 18-20 hours.

4. Separate one of the colonies grown in the medium, inoculate 10 ml of BHI liquid medium, and incubate at 37 ° C for 18-20 hours. In this way, each of the four strains is subjected to a pure separation culture (the same as in the case of the primary culture medium, except that the agar for preparing the BHI liquid medium is not added).

5. Inoculate 1% (0.1 ml) of the culture solution in the above culture medium for 1 day into 10 ml of BHI liquid medium, and incubate at 30 ° C for 18-20 hours. At this time, the above process is performed twice.

6. Prepare a medium according to the medium composition below 1 L in a 2 L beaker, and sterilize at 121 ° C for 15 minutes.

 (g / L) Soy peptide 5  Yeast extract 5  Glucose 10 KH ₂ PO ₄ 0.15 K ₂ HPO ₄ 0.15

7. 1% (10 ml) of the culture medium cultured at 5 times in the medium prepared in Table 3 is inoculated in the 1 L liquid medium and then cultured at 30 ° C for 18-20 hours. The culture medium cultured here is used as a seed culture for large-scale culture. In order to maintain the vitality of the above four strains, the strains used in the culture of BHI liquid are kept in the BHI liquid medium at least twice to maintain the viability of the seedlings. I never do that. Alternatively, when the strain is not cultured for a long time, 1 ml of the above culture medium is added to a 2 ml cryovial tube (containing 0.5 ml glycerol and 0.5 ml skim milk (10%)), thoroughly mixed, .

Experimental Example 1

Stability of new microorganisms

The safety of the novel microorganisms obtained according to the above-mentioned examples and the fermentation decaying composition containing them was confirmed without any particular problem in the experiments and researches over many years, and the stability was confirmed. The risk to humans and animals is usually the world's most reliable level 1 class of microorganisms accessible to the public without specific equipment as a result of commissioning a genetic analysis to universities using the universally common standard of bio-safety . Level 1 is a harmless microorganism that can be handled by non-specialists, and almost all of the microorganisms used in food are included.

Experimental Example 2

Evaluation of decomposition ability of food waste

To the the microorganism of 4 of the classification according to the embodiment of production species, bulk holde less Sepharose when Ah (Burkholderia cepacia), and Saccharomyces My process three Levy jiae (Saccharomyces cerevisiae) with the microorganism using the zeolite after the liquid culture, the adsorbent The present invention provides a composition for fermentation of food garbage which is prepared by a freeze-drying method which enhances viable cell count and vitality by using a method of fermentation and extinction of food waste, 2. As can be seen from the figure, 2.4% of the mass remained after about 24 hours from fermentation and decomposition of the food waste by the composition according to the present invention, showing excellent decomposition and extinguishing ability.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it will be understood by those skilled in the art that various changes and modifications may be made without departing from the spirit and scope of the invention as defined by the appended claims. Of course.

Korea Biotechnology Research Institute KCTC11085BP 20070314 Korea Biotechnology Research Institute KCTC11086BP 20070314 Korea Biotechnology Research Institute KCTC11087BP 20070314 Korea Biotechnology Research Institute KCTC11088BP 20070314

Claims (1)

Amylase, cellulase, protease and lipase enzyme having an activity of food waste decomposing activity is excellent Bacillus subtilis DYG-B14 (Bacillus subtilis DYG-B14) strain (Accession No: KCTC 11088BP) and Lactobacillus sp DYG-B13 (Lactobacillus sp. DYG-B13) strain (Accession number: featuring KCTC 11087BP) to and, characterized by a bulk holde less Sepharose cyano (Burkholderia cepacia), and Saccharomyces further arranged to further include a My three Levy process jiae (Saccharomyces cerevisiae) strain as Composition for dissolving and decomposing food waste.

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