KR101098520B1 - Microbial agent promoting composting of organic wastes treatment and manufacturing method thereof, composting method of organic wastes using the same - Google Patents

Abstract

PURPOSE: A microorganism formulation for composting organic waste is provided to ensure high decomposition activity to starch, lipid, protein, and cellulose. CONSTITUTION: A microorganism formulation for composting organic waste contains Aneurinibacillus thermoaerophilus KCCM 41604 and Thermomonospora fusca KCCM 12398. A method for preparing the microorganism formulation comprises: a step of inoculating Aneurinibacillus thermoaerophilus KCCM 41604 into a medium and culturing; a step of mixing the culture liquid and a carrier and drying; and a step of mixing with nutrient.

Description

Microbial agent promoting composting of organic wastes treatment and manufacturing method using aneuribacillus thermoaerophilus strain and its manufacturing method and composting method of the same organic wastes using the same}

The present invention relates to a microbial agent for composting organic waste, a method for preparing the same, and a method for composting organic waste using the same. More specifically, the activity of the degrading enzymes on starch, fat, protein, and cellulose is high and even at high temperatures. The present invention relates to a microbial agent having excellent composting and deodorizing ability, a method for preparing the same, and a method for composting organic waste using the same.

In general, with the increase of population, the acceleration of economic development, and the increase of national income, the standard of living of mankind is improved, while environmental pollution is accelerating. In particular, food waste, sludge from sewage and wastewater treatment plants, waste from marine processing plants, waste from agricultural and livestock processing plants, waste from food and paper mills, Increasing levels of organic waste, such as slaughterhouse slaughterhouses, livestock manure, and other organic livestock waste, are the leading sources of environmental and water pollution.

In Korea, the daily amount of food waste, a kind of organic waste, ranges from 12,000 to 20,000 tons, accounting for 23% of domestic waste. The disposal of food waste can be divided into landfill, incineration, drying, extinction through fermentation, composting and feed conversion.In the incineration process, low calorific value and high moisture content result in incomplete combustion and air pollution. There is a serious problem. The landfill treatment method generates a large amount of leachate, contaminating groundwater and soil, causing bad smell due to decay, and shortening the service life of landfills.

The fermentation annihilation method is a method in which food is completely fermented by adding food for a long time (1 month to 6 months), and moisture in the food is evaporated, and organic matter is decomposed and volatilized into carbon dioxide and water, so that the amount of food remains very small, thereby preventing the release of fermented products. Since it can be reduced, it can be said that the user's satisfaction is high. However, this method should be preceded by the development of accurate control technology for the fermentation and extinction process, and in order to secure aerobic fermentation conditions, a large amount (about 20 times the amount of food) of moisture control agent is required so that the processing equipment is enormously enlarged, resulting in food processing. The cost will be excessively high. In addition, there is a problem in that the time required for the fermentation of food is very long, economical efficiency, pollution of various various germs occurs, and the extinction performance gradually decreases, and bad odor is generated. Among these problems, the low extinction performance is due to the fact that fermentation microorganisms do not produce food degrading enzymes (amylase, protease, cellulase, lipase) properly or the activity of the produced enzymes is very low. It cannot be destroyed in time. The bad odor is because it does not effectively decompose odor-causing components of foods, such as nitrogen compounds, sulfur compounds, and lower fatty acids.

At present, the most appropriate treatment method of food waste that can supplement the above-mentioned problems can be referred to as composting. Composting refers to the formation of semi-dry solid organics in a stable form by the biological action of various aerobic microorganisms under aerobic conditions. Composting has the advantages of high organic removal rate due to the reaction at high temperature, stable processing efficiency for organic load, and no nitrification at high temperature, so that efficient use of supplied oxygen can be obtained and stable treatment can be obtained. .

Composting requires systematic research on the development of fermentation system and operation technology to improve process efficiency, and the separation of microorganisms including microorganisms and major factors necessary for composting and decomposition carriers, and microorganisms with high enzyme activity.

In composting, there is a problem of odor generation, and if the complete decomposition by microorganisms is a fundamental approach, odor can be reduced as much as possible. Most of the odor is caused by incomplete decomposition of proteins and fats in the process of degradation, so it is possible to suppress the occurrence of odor as possible by inducing complete degradation due to the active activity of proteases and lipases.

On the other hand, the composition of food waste in Korea is higher than that of Western food waste due to the high salinity of foods such as doenjang, soy sauce, kimchi, and salted fish, and the accumulation of fat components due to increased consumption of various stew, pan, fried, and meat. Activities can be difficult. The isolation of microbial strains and the formulation of microbial agents according to the characteristics of such food waste are of paramount importance.

As mentioned above, in consideration of the deodorization problem and the characteristics of the Korean food waste, the microorganism itself is important for the composting of food waste, and the potency of amylase, cellulase, protease, lipase, etc. In particular, titers such as proteases and lipases are important for suppressing the occurrence of odors. Conventionally, a large number of microorganisms are known as food fermentation strains, but the prior art has a problem in that composting efficiency is low due to low fermentation annihilation effect and deodorization effect because it does not have even decomposition activity for all starch, fat, protein and cellulose.

Therefore, the inventors of the present invention, while studying the composting of food waste can be grown at high temperature anneauribacillus thermoaerophilus, high activity of degrading enzymes for starch, fat, protein, cellulose, and the extinction rate of organic waste It confirmed that this high and came to complete this invention.

Accordingly, an object of the present invention is to provide a microbial agent having excellent high temperature fermentation and composting, deodorizing ability, and a method for preparing the same, and a method for composting organic waste using an anniuricillus thermooaerophilus microorganism.

Microbial preparation for the composting of the organic waste of the present invention for achieving the above object is characterized in that it comprises aneurinibacillus thermoaerophilus microorganisms.

The microbial agent is thermomonospora fusca ) characterized in that it further comprises a microorganism.

And the method for producing a microbial preparation for the composting of the organic waste of the present invention for achieving the above object is aneurinibacillus thermoaerophilus ( Aneurinibacillus thermoaerophilus ) culturing step of inoculating the microorganisms in the medium; A fixing step of mixing the culture solution obtained in the culturing step with a carrier and drying to obtain a microbial fixture; Characterized in that it comprises a; mixing step of mixing the nutrients that are the food ingredient of the microorganisms to the microbial fixture.

In addition, the organic waste composting method of the present invention for achieving the above object is characterized by composting the organic waste by injecting the microbial agent into the organic waste.

As described above, according to the present invention, the organic waste can be grown at a high temperature by using an anurinibacillus thermoaerophilus as a composting strain, which is capable of growing at high temperatures and has high activity of decomposition enzymes for starch, fat, protein and cellulose. In the stable fermentation can provide a microbial formulation excellent in composting, deodorizing ability.

1 is a graph showing the extinction rate of food waste using five kinds of microbial agents,
2 is a graph showing odor intensity of food waste using five kinds of microbial agents.

Hereinafter, a microbial agent for composting organic waste according to a preferred embodiment of the present invention and a manufacturing method thereof will be described in detail.

In this specification, organic wastes include food waste, sludge from sewage and wastewater treatment plants, wastes from aquatic product processing plants, wastes from agricultural and livestock processing plants, garbage from food and paper mills, slaughterhouses from slaughterhouses, livestock manure and other organic substances. It includes waste to do. Hereinafter, composting will be described as an example of the most representative food waste of organic waste.

The microbial agent of the present invention includes, for example, Aneurinibacillus thermoaerophilus . Preferably an anurinibacillus thermoaerophilus with accession number KCCM 41604 or ATCC 700303.

Other microbial agents of the present invention include Aneurinibacillus thermoaerophilus and Thermomonospora fusca microorganisms as another example. The thermosonosporus fusca is preferably Accession No. KCCM 12398, ATCC 27730.

Aneurinibacillus thermoaerophilus is characterized by high productivity of enzymes such as cellulase, amylase, protease, and lipase. Formation has high heat resistance and excellent growth at high temperatures. Actinomycetes thermomonasporus fusca is a microorganism that acts in the late stage of fermentation and plays an important role in the decomposition of hardly degradable organic substances such as cellulose and lignin.

A manufacturing method for preparing a microbial preparation is, for example, a culture step of inoculating and culturing the aneurinibacillus thermoaerophilus microorganisms in a medium, and the culture solution obtained in the culture step mixed with a carrier and dried to fix the microorganisms Obtaining a fixing step, and mixing step of mixing the nutrients of the food component of the microorganism to the microorganism fixture.

In the culture step, a medium prepared by adding tryptone, yeast extract, yeast extract, glucose, sodium chloride, NaCl, and potassium hydrogen phosphate (K ₂ HPO ₄ ) to water is used. For example, tryptone 3g, yeast extract 3g, glucose 3g, NaCl 5g, K ₂ HPO ₄ 1g is added to 1ℓ of distilled water. After inoculating the aneurinibacillus thermoaerophilus in the medium and incubated for 1 to 3 days at 40 to 60 ℃ to obtain a culture.

In the fixing step, the culture solution is mixed with a carrier and dried to obtain a microbial fixation product. This fixing step is a process of fixing the microorganisms by adsorbing the culture solution to the carrier. As the carrier, any one or more of sawdust, rice hull, and bran may be used. In addition, various known carriers capable of immobilizing microorganisms may be used. An example of the carrier may be a porous mineral, ie, pearlite, vermiculite, volcanic stone, or the like. In the fixing step, the culture medium to the carrier is preferably mixed in a volume ratio of 1 to 5:10.

After the fixing step, the nutrients are mixed in the microbial fixation, and any one or more of glucose, molasses, soy flour, ink, and fish meal may be used as nutrients. At this time, the nutrient to the microorganism fixture is preferably mixed in a volume ratio of 0.2 to 1:10.

The culturing step may further incubate thermomonasporus fusca in addition to the anurinibacillus thermoaerophilus. In this case, each medium is prepared, and then inoculated with aneurinibacillus thermoaerophilus and thermomonasporus fusca, respectively.

On the other hand, in the organic waste composting method, the above-described microbial agent is added to the organic waste to compost the organic waste. For example, organic waste is mixed at a volume ratio of 1 to 10 of the microbial preparation and fermented for 10 to 30 days to compost. At this time, the temperature may be maintained at 40 to 60 ℃, and sufficiently stirred organic waste during the fermentation period.

Hereinafter, the present invention will be described in more detail through experimental examples.

<Experimental Example 1: Searching for Optimal Strains>

As a strain for use in composting of food waste, a composting strain was developed by selecting microorganisms suitable for composting while growing at a high temperature of 40 ° C to 60 ° C.

Bacillus bacteria were selected from Aneurinibacillus thermoaerophilus (KCCM 41604), Geobacillus thermoglucosidasius (KCCM 41418), Bacillus licheniformis (KCCM 11560). It was. And actinomycetes thermomonospora fusca , KCCM 12398), Thermoactinomyces vulgaris , KCCM 41014). The microorganisms can be obtained from Korea Microorganism Conservation Center (KCCM).

Each of the Bacillus bacteria and actinomycetes was finally selected by the enzyme production test, enzyme activity test, temperature-specific culture test, pH-specific growth test, flame resistance test, and the extinction performance and odor of food waste for these strains The intensity of development was tested.

1. Investigation of enzyme activity of microorganism

Since food waste consists of carbohydrates, proteins, fats, and fibrin, it is known that anniuribacillus thermoaerophilus (hereinafter referred to as KCCM 41604) and geo Bacillus thermoglucosidosis (hereinafter referred to as KCCM 41418), Bacillus rickenformis (hereinafter referred to as KCCM 11560), thermomonasporus fusca (hereinafter referred to as KCCM 12398) and thermoactinomyces vulgaris (hereinafter referred to as KCCM 41014) microorganisms were investigated.

Carbohydrate degrading enzyme (amylase), proteolytic enzyme (protease), lipolytic enzyme (lipase), fibrinolytic enzyme (cellulase) method of investigation of the activity is as follows, the results are shown in Table 1 below.

(1) measuring amylase activity

The medium used to confirm the starch resolution of the strain was a 1% soluble starch added to LB agar medium (Tryptone; 10g, Yeast extract; 5g, NaCl; 5g, Agar; 15g, distilled water; 1L) After inoculating the strain and incubating at 50 ° C. for 24 hours, the activity was measured by observing the clear zone size generated by staining with iodine solution (iodine 1g, potassium iodide 2g, distilled water 300ml).

(2) measuring protease activity

The medium used to confirm the protein resolution of the strain was used to add 1% skim milk to LB agar medium, the activity was measured by inoculating the strain and incubated at 50 ℃ for 24 hours and observed the size of the resulting clear zone. .

(3) lipase activity measurement

The medium used to confirm the fat resolution of the strain was used to add 1.0% of tributyrin to LB agar medium, incubated for 24 hours at 50 ℃ to observe the resulting clear zone size Activity was measured.

(4) measuring cellulase activity

The medium used to confirm the cellulolytic resolution of the strain was the addition of 0.5% carboxy methyl cellulose (CMC) to the LB agar medium, inoculated with the strain and incubated at 50 ℃ for 24 hours, 0.2% Congo red (congo red) Activity was measured by observing the clear zone size generated when staining with 30 minutes and washed with 1M NaCl for 15 minutes.

Fungus
Strain name
Enzyme activity (diameter of clear zone, mm) Amylase Protease Lipase Cellulase
Germ
KCCM 41604 15 21 8 14 KCCM 41418 29 18 14 22 KCCM 11560 16 13 11 19 Actinomycetes
KCCM 12398 18 12 5 27 KCCM 41014 13 7 0 13

Referring to Table 1, the enzyme activity shows a lot of difference according to the species. Among the strains, KCCM 41418 was highest for amylase and lipase activity, KCCM 41604 for protease, and KCCM for cellulase. 12398 showed the highest activity.

From the above results, it can be seen that the size of the clear zone for each enzyme varies depending on the type of microorganism. This means that there is a big difference in enzyme production capacity and activity according to the type of microorganism, and eventually there are many differences in food extinction ability according to the species.

2. Investigation of growth of microorganisms by temperature

In order to determine the microbial growth temperature range, the strains were inoculated with LB agar medium and incubated at 20, 30, 40, 50, 60, 70 ° C for 48 hours, and the growth of the bacteria was observed. Cultureability was evaluated by the method of sensory investigation of the size of the strain colony and the results are shown in Table 2 below.

Fungus
Strain name
Growth growth by temperature 20 ℃ 30 ℃ 40 ℃ 50 ℃ 60 ℃ 70 ℃
Germ
KCCM 41604 - + ++ +++ ++ - KCCM 41418 - + +++ +++ ++ - KCCM 11560 ++ ++ +++ + - - Actinomycetes
KCCM 12398 + ++ +++ +++ + - KCCM 41014 + ++ +++ ++ - -

(+++: Excellent, ++: Normal, +: Weak,-: Impossible)

Referring to Table 2, KCCM 41604 and KCCM 41418 showed excellent growth at 40 ~ 60 ℃, KCCM 12398 and KCCM 41014 grew well at 40 ~ 50 ℃. On the other hand, KCCM 11560 was weak growth at 50 ℃, it was well grown at 30 ~ 40 ℃. Therefore, other microorganisms except KCCM 11560 were found to be applicable as a composting microbial agent because of their high growth at high temperature of 40 ~ 60 ℃.

3. Investigate the growth of microorganisms by pH

In order to determine the pH range of microorganisms, strains were inoculated into the LB agar medium, and cultured at 45 ° C. for 48 hours in a range of pH 4 to 10, and the growth of the bacteria was observed. Cultureability was evaluated by the method of sensory investigation of the size of the colony and the results are shown in Table 3 below.

Fungus
Strain name
pH-specific growth pH4 pH5 pH6 pH7 pH8 pH9 pH10
Germ
KCCM 41604 - + ++ +++ +++ + - KCCM 41418 - + ++ +++ ++ ++ - KCCM 11560 - ++ ++ +++ ++ - - Actinomycetes
KCCM 12398 - - ++ +++ ++ - - KCCM 41014 - - - +++ ++ + -

(+++: Excellent, ++: Normal, +: Weak,-: Impossible)

Referring to Table 3, KCCM 11560 exhibited excellent growth at pH 5-8, and grew relatively well in acid, and KCCM 12398 and KCCM 41014 grew well at neutral pH 6-8. On the other hand, KCCM 41604 and KCCM 41418 grew well in a very wide pH range of pH 5-9 and were found to be suitable as composting microbial agents.

4. Flame resistance investigation of microorganisms

To determine the growth of microorganisms according to salinity, 1%, 2%, 4%, 6%, 8% NaCl (w / v) was added to LB agar medium, respectively, and then inoculated with bacteria and incubated at 50 ° C for 24 hours. After the growth of the bacteria was observed. Cultureability was evaluated by the method of sensory investigation of the size of the strain colony and the results are shown in Table 4 below.

Fungus
Strain name
Growth growth by salinity concentration Salinity 1% Salinity 2% Salinity 4% Salinity 6% Salinity 8%
Germ
KCCM 41604 +++ +++ ++ + - KCCM 41418 +++ +++ ++ + - KCCM 11560 +++ +++ ++ - - Actinomycetes
KCCM 12398 +++ +++ + - - KCCM 41014 +++ ++ + - -

(+++: Excellent, ++: Normal, +: Weak,-: Impossible)

From the results of Table 4, KCCM 41604 and KCCM 41418 are able to grow up to 1% to 6% of salt concentration, indicating that they have strong resistance to salts, and thus can be applied as composting microbial preparations of food wastes with high variation in salt concentration. Could. On the other hand, KCCM 11560, KCCM 12398 and KCCM 41014 were able to grow up to 1% to 4% salt concentration.

5. Investigation of culture yield of microorganism

In order to investigate the culture yield of the microorganisms, the culture yield was examined by culturing the microorganisms in LB medium for 24 hours at 50 ° C. and transferring them to LB agar plate medium for 2 days and measuring the number of colonies formed. The results are shown in Table 5 below.

Fungus Strain name Culture yield (× 10 ⁶ CFU / ml)
Germ
KCCM 41604 752 KCCM 41418 831 KCCM 11560 593 Actinomycetes
KCCM 12398 94 KCCM 41014 63

Referring to Table 5, KCCM 41604 and KCCM 41418 showed high culture yields in the bacteria, and KCCM 12398 showed high culture yields in actinomycetes.

6. Investigation of culture in food eluate and solids

Food waste can be divided into elutable portions containing most of the moisture and other solids (including components adsorbed on the solids) in terms of their physical properties. The initial stage of food waste composting is the dissolution of the elutable portion, and the later stage is the decomposition of the solid portion.

In order to evaluate the degradability of the elutable component and the solid component of the microorganism, the test was carried out as follows. In other words, by adding an appropriate amount of water (half of the weight of food waste) to food waste collected in an apartment complex in Gwangju city and adding agar to the elutable portion obtained by gentle stirring, a solid medium for microbial culture was prepared and sterilized. Colonies of the resulting strains were inoculated with each microorganism and incubated at 50 ° C. for 2 days.

On the other hand, in order to evaluate the degradability of the solid part of the food, after dehydration and washing the food solids to high-speed crushing to make a porridge state (slurry state), 5 times distilled water is added in the weight ratio of solids and sterilized by adding agar. Thereafter, each microorganism was inoculated and cultured at 50 ° C. for 2 days, and the number of colonies of the resulting strains was measured. The results are shown in Table 6 below.

Fungus
Strain name
Culture yield from food waste (× 100,000 CFU / ml) Elutable part Solid part
Germ
KCCM 41604 362 107 KCCM 41418 578 216 KCCM 11560 225 78 Actinomycetes
KCCM 12398 82 29 KCCM 41014 47 22

As shown in Table 6, among the bacteria, KCCM 41604 and KCCM 41418 had excellent cultivation in both elution and solids, and in actinomycetes, KCCM 12398 had excellent cultivation in both parts, which could effectively decompose food. It has been shown to have excellent properties.

Experimental Example 2 Evaluation of the Extinction Rate of Food Waste

Finally, five kinds of microbial preparations were prepared using KCCM 41604, KCCM 41418, KCCM 11560, KCCM 12398, and KCCM 41014 to evaluate the extinction rate of food waste.

To prepare the microbial preparation, each strain of 5 liters of nutrient broth medium (Tryptone 3g, yeast extract 3g, glucose 3g, NaCl 5g, K ₂ HPO ₄ 1g, H ₂ O 1L) was inoculated with each strain. Liquid cultures were obtained at 50 ° C. for 48 hours. Sawdust, which is a culture medium to a carrier, was mixed in a volume ratio of 2:10 and dried to form a microorganism. And a microbial preparation for food waste compost was prepared by mixing a nutrient-to-microbial dry matter, in which the glucose, molasses, soy flour, salt and fish meal were mixed in the same ratio in a volume ratio of 0.5: 10.

Five kinds of prepared microbial preparations were fermented by adding them to a food waste composting apparatus with a total capacity of 20 liters equipped with a stirrer, a temperature controller, a deodorizer and a fan. The internal temperature of the apparatus was maintained at 50 ° C. during the 30 day fermentation period, the stirring cycle was 20 minutes per hour, and the stirring speed was 6 revolutions per minute. 2 kg of food waste (food waste discharged from Gwangju downtown apartment complex, average water content of 80%) was investigated every day, and the extinction rate was examined, and the results are shown in FIG. 1.

Referring to FIG. 1, KCCM 41604 exhibited the highest extinction rate with an extinction rate of 81% to 93% and an average extinction rate of 88.6% for 30 days. Next, KCCM 41418 was 87.0%, KCCM 11560 was 80.2%, KCCM 12398 was 79.2%, and KCCM 41014 was 77.5%.

Experimental Example 3 Measurement of Odor Level of Food Waste

In order to determine the degree of odor generated during the composting of food waste by using the microbial agent, the sensory results of five sensory testers with healthy noses during the experiment of the second experimental example were directly quantified by relative indicators 1 to 5. (Odor intensity 1: no smell at all, odor intensity 2: odor slightly south, odor intensity 3: odor can be felt clearly, odor strength 4: unpleasant odor, odor 5: very unpleasant smell Severe) and the average value thereof was investigated and the results are shown in FIG. 2.

Referring to FIG. 2, KCCM 41604 exhibited a odor strength of 2.4 at a odor strength of 1 for 30 days in the composting period, and was found to have the lowest odor. In addition, KCCM 12398> KCCM 41014> KCCM 11560> KCCM 41418 showed strong odor.

As such, the aneurinibacillus thermoaerophilus was confirmed that the food waste cultivation, that is, the food extinction performance is the best and the odor is very good compared to other microorganisms. Therefore, it was confirmed that the microorganism formulation of the present invention, Aniurib Bacillus thermoaerophilus is very excellent in food extinction performance and very low odor inducing strength can effectively compost food without causing odor.

In the first experimental example, the aneurinibcillus thermoaerophilus and geobacillus thermoglucosidus showed relatively high activity in the enzyme activity, grew well at a high temperature of 40 to 60 ℃ and other food waste fractions. It showed higher culture than microorganisms. In particular, the aneurinibacillus thermoaerophilus showed the highest efficiency in the extinction rate of food waste, the lowest in the odor induction intensity, and therefore appeared to be most suitable as a composting microbial preparation of food waste.

While the present invention has been described with reference to exemplary embodiments, it is to be understood that the invention is not limited to the disclosed exemplary embodiments.

Therefore, the true scope of protection of the present invention should be defined only by the appended claims.

Claims (4)

Aneurinibacillus thermoaerophilus KCCM 41604 A microbial agent for the composting of organic waste, characterized in that it comprises a microorganism. The microbial preparation of claim 1, wherein the microbial preparation further comprises Thermomonospora fusca KCCM 12398 microorganisms. Aneurinibacillus thermoaerophilus KCCM 41604 microorganisms are cultured by inoculating the culture medium;
A fixing step of mixing the culture solution obtained in the culturing step with a carrier and drying to obtain a microbial fixture;
Mixing step of mixing the nutrients as a food ingredient of the microorganisms to the microbial fixture; manufacturing method of the microbial preparation for the composting of the organic waste comprising a. A method of composting an organic waste, characterized in that the organic waste is composted by injecting the microbial agent of claim 1 or 2 into the organic waste.

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