Strain for degradation of organic waste and method for treating organic waste using same

Abstract

The present disclosure relates to a strain for degradation of organic waste and a method for treating organic waste using same. The strain according to embodiments of the present application exhibits excellent enzymatic activity for degrading starch, protein, and fat even in high-temperature environments and in the presence of high concentrations of salt, and can effectively degrade limit dextrin and dextran.

Description

A strain for decomposing organic waste and a method for treating organic waste using the strain

[0001] This invention relates to a strain for decomposing organic waste and a method for treating organic waste using the strain.

[0002] Large quantities of food waste are typically generated from households and restaurants. Despite being recyclable organic waste, most food waste is disposed of through incineration or landfill. Due to its high moisture content, incineration requires significant fuel consumption, leading to high disposal costs. Furthermore, the release of carbon dioxide and dioxins can negatively impact the atmospheric environment. Landfilling food waste also presents challenges, such as the foul odor generated during the decomposition process. Furthermore, the limited landfill space required for the disposal limits the volume that can be processed.

[0003] Due to these issues, research is being conducted on ways to efficiently utilize organic waste, such as food waste, without incineration or landfilling. In particular, active research is being conducted on utilizing microorganisms to compost organic waste, which could be utilized in fields such as agriculture and reduce environmental pollution, achieving the dual benefit of reducing environmental pollution. In particular, for Korea, a resource-poor country, policies that safely recycle organic waste into resources without negatively impacting the environment are crucial.

[0004] Food waste, a representative type of organic waste, is recycled into feed, fertilizer (compost), and bioenergy. Most of it is animal feed or agricultural fertilizer (compost). Food waste is primarily generated from group cafeterias in homes, restaurants, schools, military bases, and businesses, and consists of leftover food (30%), waste from food ingredient distribution and cooking (57%), food ingredients discarded in storage (9%), and uneaten food (4%). The amount, nature, degree of decay, and presence of foreign substances in sanitary management conditions vary depending on the source. However, because these wastes are mixed and recycled without being separated, concerns about the spread of livestock infectious diseases through feed and fertilizer, as well as soil and water contamination, many people are reluctant to use them.

[0005] [Prior Art Literature]

[0006] [Patent Document]

[0007] Chinese Patent Publication No. 107537842.

[0008] The present invention aims to provide a strain for decomposing organic waste and a method for treating organic waste using the strain.

[0009] However, the problems that the present invention seeks to solve are not limited to the problems mentioned above, and other problems not mentioned will be clearly understood by those skilled in the art from the description below.

[0010] The first aspect of the present invention provides a Bacillus atrophaeus YBK-W4 (KCTC 15572BP) strain characterized by decomposing organic waste.

[0011] The second aspect of the present invention provides a method for treating organic waste, comprising: culturing a strain according to the first aspect to obtain a culture solution; and mixing the culture solution with organic waste to convert the organic waste into water.

[0012] The third aspect of the present invention provides an organic waste treatment system using a strain according to the first aspect.

[0013] The strain according to the embodiments of the present invention can decompose organic waste into water.

[0014] The strain according to the embodiments of the present invention not only has excellent enzyme activity for decomposing starch, protein, and fat even in a high-temperature environment and in the presence of high concentrations of salt, but can also decompose limit dextrin and dextran well.

[0015] The method for treating organic waste according to the embodiments of the present invention can treat organic waste within about 24 hours.

[0016] The organic waste decomposition rate of the organic waste treatment method according to the embodiments of the present invention may be about 90% or more, about 95% or more, or about 98% or more.

[0017] The method for treating organic waste according to the embodiments of the present invention can be an environmentally friendly process that emits almost no heavy metals.

[0018] The method for treating organic waste according to the embodiments of the present invention may be an environmentally friendly process by converting organic waste into water.

[0019] The method for treating organic waste according to the embodiments of the present invention may be an economical process.

[0020] Figure 1 is a growth curve showing changes in pH, dissolved oxygen (DO), and optical density (OD) at 660 nm over time when the YBK-W4 strain was cultured at 47°C in one embodiment of the present invention.

[0021] Figure 2 is an image of a clear zone for analyzing the starch decomposition ability of the YBK-W4 strain in one embodiment of the present invention.

[0022] Figure 3 is an image of a transparent ring for analyzing the protein decomposition ability of the YBK-W4 strain in one embodiment of the present invention.

[0023] Figure 4 is an image of a transparent ring for analyzing the fat decomposition ability of the YBK-W4 strain in one embodiment of the present invention.

[0024] FIG. 5 is an image of a transparent ring for analyzing the dextran decomposition ability of the YBK-W4 strain in one embodiment of the present invention.

[0025] Figure 6 is an image of a transparent ring for analyzing the limit dextrin decomposition ability of the YBK-W4 strain in one embodiment of the present invention.

[0026] Figure 7 shows the 16s rRNA base sequence of the YBK-W4 strain in one embodiment of the present invention.

[0027] Hereinafter, with reference to the attached drawings, implementation examples and embodiments of the present invention will be described in detail so that those skilled in the art can easily practice the present invention. However, the present invention may be implemented in various different forms and is not limited to the implementation examples and embodiments described herein. In addition, in the drawings, parts irrelevant to the description have been omitted to clearly explain the present invention, and similar parts have been designated with similar drawing reference numerals throughout the specification.

[0028] Throughout this specification, when a part is said to be "connected" to another part, this includes not only cases where it is "directly connected" but also cases where it is "electrically connected" with another element in between.

[0029] Throughout this specification, when it is said that an element is "on" another element, this includes not only cases where the element is in contact with the other element, but also cases where another element exists between the two elements.

[0030] Throughout this specification, whenever a part is said to "include" a component, this does not mean that it excludes other components, but rather that it may include other components, unless otherwise specifically stated.

[0031] The terms "about," "substantially," and the like used in this specification are used in a meaning that is at or close to the numerical value when manufacturing and material tolerances inherent in the meanings mentioned are presented, and are used to prevent unscrupulous infringers from unfairly exploiting the disclosure in which exact or absolute values ​​are mentioned to aid understanding of the present application.

[0032] The terms “step of ~” or “step of ~” as used throughout this specification do not mean “step for ~.”

[0033] Throughout this specification, the term "combination(s) thereof" included in the expressions in the Makushi format means one or more mixtures or combinations selected from the group consisting of the components described in the expressions in the Makushi format, and means including one or more selected from the group consisting of said components.

[0034] Throughout this specification, references to “A and / or B” mean “A or B, or A and B.”

[0035] Below, the implementation examples of the present invention are described in detail, but the present invention may not be limited thereto.

[0036] The first aspect of the present invention provides a Bacillus atrophaeus YBK-W4 (KCTC 15572BP) strain characterized by decomposing organic waste.

[0037] The second aspect of the present invention provides a method for treating organic waste, comprising: culturing a strain according to the first aspect to obtain a culture solution; and mixing the culture solution with organic waste to convert the organic waste into water.

[0038] Detailed descriptions of parts that overlap with the first aspect of the present application have been omitted, but the contents described in the first aspect of the present application may be equally applied even if the description is omitted in the second aspect of the present application.

[0039] In one embodiment of the present invention, the culturing may be performed at a temperature range of about 20°C to about 80°C, but may not be limited thereto. In one embodiment of the present invention, the culturing may be performed at a temperature range of about 20°C to about 80°C, about 20°C to about 70°C, about 20°C to about 60°C, about 30°C to about 80°C, about 30°C to about 70°C, about 30°C to about 60°C, about 40°C to about 80°C, about 40°C to about 70°C, or about 40°C to about 60°C, but may not be limited thereto. In one embodiment of the present invention, the culturing may be performed at about 50°C.

[0040] In one embodiment of the present invention, the culturing may be performed for about 24 hours to about 72 hours, but may not be limited thereto. In one embodiment of the present invention, the culturing may be performed for about 24 hours to about 72 hours, about 24 hours to about 66 hours, about 24 hours to about 60 hours, about 24 hours to about 54 hours, about 30 hours to about 72 hours, about 30 hours to about 66 hours, about 30 hours to about 60 hours, about 30 hours to about 54 hours, about 36 hours to about 72 hours, about 36 hours to about 66 hours, about 36 hours to about 60 hours, about 36 hours to about 54 hours, about 42 hours to about 72 hours, about 42 hours to about 66 hours, about 42 hours to about 60 hours, or about 42 hours to about 54 hours, but may not be limited thereto. In one embodiment of the present invention, the culturing may be performed for about 48 hours.

[0041] In one embodiment of the present invention, the culture solution may be used by being adsorbed to a carrier.

[0042] In one embodiment of the present invention, the carrier may include sawdust and bamboo pieces.

[0043] In one embodiment of the present invention, the sawdust may be included in an amount of about 10% to about 80% by volume of the carrier, but may not be limited thereto. In one embodiment of the present invention, the sawdust may be included in an amount of about 10% to about 80% by volume, about 10% to about 75% by volume, about 10% to about 70% by volume, about 20% to about 80% by volume, about 20% to about 75% by volume, about 20% to about 70% by volume, about 30% to about 80% by volume, about 30% to about 75% by volume, about 30% to about 70% by volume, about 40% to about 80% by volume, about 40% to about 75% by volume, about 40% to about 70% by volume, about 50% to about 80% by volume, about 50% to about 75% by volume, or about 50% to about 70% by volume of the carrier, but may not be limited thereto. In one embodiment of the present invention, the sawdust may be included in about 60% by volume of the carrier.

[0044] In one embodiment of the present invention, the bamboo pieces may be included in about 20% to about 70% by volume of the carrier, but may not be limited thereto. In one embodiment of the present invention, the bamboo pieces may be included in about 20% to about 70% by volume of the carrier, about 20% to about 65% by volume, about 20% to about 60% by volume, about 20% to about 55% by volume, about 20% to about 50% by volume, about 30% to about 70% by volume, about 30% to about 65% by volume, about 30% to about 60% by volume, about 30% to about 55% by volume, or about 30% to about 50% by volume of the carrier, but may not be limited thereto. In one embodiment of the present invention, the bamboo pieces may be included in about 40% by volume of the carrier.

[0045] In one embodiment of the present invention, the size of the bamboo piece may be about 1 cm to about 50 cm in width and about 5 cm to about 40 cm in length.

[0046] In one embodiment of the present invention, the adsorption may be performed at a temperature range of about 20°C to about 80°C, but may not be limited thereto. In one embodiment of the present invention, the adsorption may be performed at a temperature range of about 20°C to about 80°C, about 20°C to about 70°C, about 20°C to about 60°C, about 30°C to about 80°C, about 30°C to about 70°C, about 30°C to about 60°C, about 40°C to about 80°C, about 40°C to about 70°C, or about 40°C to about 60°C, but may not be limited thereto. In one embodiment of the present invention, the adsorption may be performed at about 50°C.

[0047] In one embodiment of the present invention, the adsorption may be performed for about 12 hours to about 72 hours, but may not be limited thereto. In one embodiment of the present invention, the adsorption is performed for about 12 hours to about 72 hours, about 12 hours to about 66 hours, about 12 hours to about 60 hours, about 12 hours to about 54 hours, about 12 hours to about 48 hours, about 24 hours to about 72 hours, about 24 hours to about 66 hours, about 24 hours to about 60 hours, about 24 hours to about 54 hours, about 30 hours to about 72 hours, about 30 hours to about 66 hours, about 30 hours to about 60 hours, about 30 hours to about 54 hours, about 36 hours to about 72 hours, about 36 hours to about 66 hours, about 36 hours to about 60 hours, about 36 hours to about 54 hours, about 42 hours to about 72 hours, about 42 hours to about 66 hours, about 42 The adsorption may be performed for, but is not limited to, about 60 hours, or about 42 hours to about 54 hours. In one embodiment of the present invention, the adsorption may be performed for about 48 hours.

[0048] In one embodiment of the present invention, converting the organic waste into water may be performed at a temperature range of about 20°C to about 80°C, but may not be limited thereto. In one embodiment of the present invention, converting the organic waste into water may be performed at a temperature range of about 20°C to about 80°C, about 20°C to about 70°C, about 20°C to about 60°C, about 30°C to about 80°C, about 30°C to about 70°C, about 30°C to about 60°C, about 40°C to about 80°C, about 40°C to about 70°C, or about 40°C to about 60°C, but may not be limited thereto. In one embodiment of the present invention, converting the organic waste into water may be performed at about 50°C.

[0049] In one embodiment of the present invention, the method for treating organic waste may include the following processes 1) to 4):

[0050] 1) Bacillus atrophaeus YBK-W4 (KCTC 15572BP) strain is cultured in liquid form in an incubator at an internal temperature of 50℃ for 48 hours;

[0051] 2) Add the above culture solution to the carrier and stir at 50°C for 48 hours;

[0052] 3) The above strain and the carrier are placed in a reaction vessel and stirred at 50°C with an internal stirrer. At this time, air is supplied to the interior of the reaction vessel, water vapor inside the reaction vessel is discharged, water is discharged through a condenser, and the remaining air is discharged through a deodorizer.

[0053] 4) Organic waste is decomposed into water within 3 to 4 hours in the reaction vessel, the decomposed water is discharged as steam within 24 hours, and the residue is discharged every 1 to 3 months.

[0054] In one embodiment of the present invention, the decomposition principle of the organic waste may include the following 1) to 4):

[0055] 1) After the secured original bacteria are anaerobically cultured for 48 hours to create a culture solution, the culture solution and carrier are placed in an organic waste treatment device and stirred at 50°C for 48 hours to evenly adsorb the culture solution onto the carrier;

[0056] 2) Organic waste is fed into the organic waste treatment device, and the appropriate temperature and appropriate oxygen are provided to convert the organic waste into water (H2O) through enzymes produced through the metabolic action of microorganisms;

[0057] 3) When the high-pressure oxygen newly supplied to the organic waste treatment device collides with the low-pressure internal water vapor, the oil vapor and water vapor are separated, and only the water vapor is discharged;

[0058] 4) The discharged water vapor is discharged as clean water through the condenser, and the remaining dry air can be passed through a microbial deodorizer to prevent bad odor.

[0059] The third aspect of the present invention provides an organic waste treatment system using a strain according to the first aspect.

[0060] Detailed descriptions of overlapping parts with the first and second aspects of the present application have been omitted, but the contents described for the first and second aspects of the present application may be equally applied even if the description is omitted in the third aspect of the present application.

[0061] In one embodiment of the present invention, the organic waste treatment system may include a culture medium and a reaction vessel, wherein the strain is cultured in a liquid in the culture medium to obtain a culture solution, and the culture solution and organic waste may be mixed in the reaction vessel and the organic waste may be converted into water.

[0062] In one embodiment of the present invention, the reaction vessel includes a stirring blade, and the stirring blade may be used to mix the culture solution and the organic waste.

[0063] In one embodiment of the present invention, the organic waste treatment system may further include a deodorizing system.

[0064] In one embodiment of the present invention, the deodorizing system may include spraying the strain according to claim 1 into the steam from which the organic waste is converted and evaporated.

[0065] Hereinafter, the present invention will be described in more detail using examples. However, the following examples are provided only to help understand the present invention, and the contents of the present invention are not limited to the following examples.

[0066] <Strain Deposit>

[0067] Bacillus atrophaeus YBK-W4 (KCTC 15572BP) strain was deposited at the Korean Collection for Type Cultures (KCTC) of the Korea Research Institute of Bioscience and Biotechnology on August 29, 2023, and was assigned the accession number KCTC 15572BP.

[0068] <Evaluation of the food decomposition ability of the strain>

[0069] In order to isolate a strain that can grow at high temperatures and decompose various nutrients, dextran, and limit dextrin, 100 g of the lumps present in the applicant's large-scale food waste disposer were suspended in 1 L of sterile saline solution and serially diluted. The resulting mass was spread on a selective agar medium and cultured at 50°C. The grown colonies were toothpicked onto a high-salinity medium to select colonies that grew well in the first round, and strains with excellent dextran decomposition ability were selected in the second round. Among the selected strains, the strain with the best organic matter decomposition ability was selected in the third round. Among the strains selected in the third round, strains YBK-W1, YBK-W2, YBK-W3, YBK-W4, and YBK-W5 that grew well even at 50°C were selected. The activities of various decomposition enzymes of the above strains are as shown in Table 1 below:

[0070] Strain Amylase Protease Lipase Cellulase Dextranase Limit dextrin B. subtilis YBK-1KCTC 1530+++++++++ B. licheniformis W-2+++++++ B. stearothermophilis W-3++ B. atrophaeus W-4++++++++++++++++++++++++++++ B. amyloliquefaciens W-5++++++++++++

[0071] +++++: Very high ++++: Excellent +++: Average ++: Weak + : Very weak

[0072] Referring to Table 1 above, all five strains produced six types of enzymes, but the YBK-W4 strain was found to be the best.

[0073] 1. Salt resistance evaluation

[0074] After adding 0% to 12% sodium chloride (NaCl) to LB medium, the culture was incubated at 37°C for 24 hours, and the grown colonies were selected. The results of the salt tolerance evaluation are shown in Table 2 below:

[0075] NaCl concentrationYBK-1YBK-W2YBK-W3YBK-W4YBK-W50++++++++++++++++++3++++++++++++++6+++++++9+++12++

[0076] +++++: Very high ++++: Excellent +++: Average ++: Weak + : Very weak

[0077] Referring to Table 2 above, the YBK-W4 strain grew very vigorously in a liquid medium containing 12% NaCl in LB medium. Among the strains, YBK-W3 had the weakest salt tolerance.

[0078] 2. Growth curve (Figure 1)

[0079] When the YBK-W4 strain was cultured at 47°C in modified M9 minimal medium, changes in pH, DO, and OD values ​​over time were observed, and the YBK-W4 strain was cultured at 45°C to obtain a growth curve.

[0080] Time (minutes)pHDOOD016.42100.00.6926.41100.00.6936.47100.00.6946.47100.01.1356.4398.02.2866.4772.03.5076.4758.85.1186.6545.56.5496.6530.07.68106.6812.08.83 116.513.69.48126.40-10.13136.460.610.54146.420.810.89156.6625.111.01166.6066.811.01176.7886.911.01187.1893.811.09197.52100.011.09207.75100.011.09

[0081] Referring to Fig. 1 and Table 3 above, the YBK-W4 strain showed vigorous growth, showing maximum absorbance at 15 hours, similar to 37°C, even at a high temperature of 45°C.

[0082] 3. Resolution Evaluation

[0083] 3-1. Starch decomposition ability (Fig. 2)

[0084] The strain isolated was toothpicked onto an agar medium containing 1% starch in LB medium, cultured at 37°C for 24 hours, and then stained with a 0.1 N iodine solution to measure the size of the clear zone.

[0085] 3-2. Protein degradation (Figure 3)

[0086] Protease secretion ability was measured by toothpicking on skim milk agar medium and measuring the size of the transparent ring formed after culturing at 37°C for 24 hours.

[0087] 3-3. Fat decomposition ability (Fig. 4)

[0088] Toothpicking was performed on a lipase isolation agar medium supplemented with 1% tributyrin for the lipase isolation assay, and the size of the formed transparent ring was measured after culturing at 37°C for 24 hours.

[0089] 3-4. Dextran resolution (Figure 5)

[0090] Dextran degradation experiments were performed by toothpicking on M9 minimal agar medium supplemented with 1% dextran as the sole carbon source, and culturing at 37°C for 24 hours, and measuring the size of the formed pellicle.

[0091] 3-5. Limit dextrin resolution (Fig. 6)

[0092] Limit dextrin resolution experiments were conducted by toothpicking on M9 minimal agar medium supplemented with 1% limit dextrin as the sole carbon source, and measuring the size of the formed transparent ring after incubation at 37°C for 24 hours.

[0093] 4. Preparation of enzyme solution

[0094] Crude enzyme solution for measuring amylase enzyme activity was prepared by culturing the culture medium for 48 hours at 37℃ with 0.2% soluble starch as the sole carbon source in M9 minimal medium, centrifuging the supernatant. The crude enzyme solution for dextranase and limit dextrinase activities was prepared by using the supernatant cultured in the same manner as the amylase crude enzyme solution, with 0.2% dextran and limit dextrin added as carbon sources. The supernatant cultured in M9 medium with 0.5% olive oil added was used for measuring lipase activity. The supernatant cultured in minimal medium with 1% gelatin as the sole nitrogen source for protease activity was used. Protein content was quantified using a Bradford assay kit.

[0095] 4-1. Amylase activity

[0096] Amylase enzyme activity was analyzed using the total carbohydrate assay kit (The total carbohydrate assay kit; Sigma, MAK104-1KT).

[0097] Temperature (℃) Amylase activity (U / mL) Relative activity of amylase (%) 322.1858373.5293423.78100473.5393503.3789553.1182602.5166651.5240

[0098] Referring to Table 4 above, the optimum temperature for amylase enzyme activity of the YBK-W4 strain was 42°C, and it was confirmed that it had very strong thermal stability, as it exhibited 66% activity even at 60°C.

[0099] pH Amylase Activity (U / mL)Relative Amylase Activity (%)3.02.18584.03.11825.03.48926.03.53937.03.781008.03.21849.02.5166

[0100] Referring to Table 5 above, the optimal pH of amylase of the YBK-W4 strain was 7.0, and it showed 66% activity even at pH 9.0.

[0101] Salinity of NaCl (%) Activity of Amylase (U / mL) Relative Activity of Amylase (%) 03.78 100 3.03.76 99 6.03.33 88 9.02.29 60 12.00.90 24

[0102] Referring to Table 6 above, the amylase enzyme of the YBK-W4 strain showed 60% enzyme activity at 9% salinity and 24% enzyme activity at 12% salinity.

[0103] 4-2. Protease activity

[0104] Protease activity was assayed using the Pierce Fluorescent Protease Assay Kit (23266, ThermoFisher Scientific).

[0105] Temperature (℃)Protease activity (U / mL)Relative protease activity (%)323.6274374.89100424.5893474.2186503.3368552.9961602.5151652.1243

[0106] Referring to Table 7 above, the optimal temperature for protease enzyme activity of the YBK-W4 strain was 37°C, and 43% activity was observed even at 65°C.

[0107] pHProtease activity (U / mL)Relative activity of protease (%)2.02.08433.03.61744.03.98815.04.13846.04.57937.04.891008.02.51669.02.0141

[0108] Referring to Table 8 above, the optimal pH of the protease of the YBK-W4 strain was 7.0, and it showed 41% activity even at pH 9.0.

[0109] 4-3. Lipase activity

[0110] Lipase activity was measured using a lipase activity assay kit (CAS number MAK046, Sigma-Aldrich). The combined enzyme reaction was measured colorimetrically (at 570 nm), which was proportional to the enzyme activity present in the sample. Lipase enzyme activity was measured as the increase in 570 nm light in real time over the reaction time.

[0111] Temperature (℃) Activity of Lyphase (U / mL) Relative Activity of Lyphase (%) 323.6889374.11100423.9897473.5386503.3782553.1376602.5161651.5237

[0112] Referring to Table 9 above, the optimal temperature for the life phase of the YBK-W4 strain was 37°C, and it showed 37% activity even at 65°C.

[0113] Activity of pH-Lyphace (U / mL)Relative activity of Lyphace (%)3.01.11274.02.27555.03.18776.03.54867.04.111008.03.14769.02.5161

[0114] Referring to Table 10 above, the optimal pH of the YBK-W4 strain was 7.0, and it showed 41% activity even at pH 9.0.

[0115] 4-4. Dextranase activity

[0116] Dextranase activity was analyzed by adding 0.1 mL of crude enzyme solution to 0.5 mL of dextran (T5,000-40,000) solution, 0.1 M citrate, 0.2 M Na2HPO4, and 0.4 mL of buffer solution, reacting at 37°C, and then quantifying reducing sugar using the DNS method.

[0117] Temperature (℃) Activity of dextranase (U / mL) Relative activity of dextranase (%) 321.75 60 372.29 79 422.89 100 472.57 89 50 2.02 70 55 1.77 6 16 0 1.10 38 6 5 0.81 28

[0118] Referring to Table 11 above, the optimal temperature for the YBK-W4 strain was 42°C, and it showed 28% activity even at 65°C.

[0119] pH Activity of dextranase (U / mL) Relative activity of dextranase (%) 3.0 0.9 3 3 2 4.01 14 39 5.02 02 7 06.02 55 887.02 89 1008.02 41 839.00 91 31

[0120] Referring to Table 12 above, the optimal pH of the YBK-W4 strain was 7.0, and it showed 31% activity even at pH 9.0.

[0121] Salinity of NaCl (%) Activity of dextranase (U / mL) Relative activity of dextranase (%) 02.89 1003.02.8 1976.02.18 759.01.19 4112.00.10 19

[0122] Referring to Table 13 above, the dextranase enzyme of the YBK-W4 strain showed 41% activity even at 9% salinity.

[0123] 4-5. Limit dextrin enzyme activity

[0124] Limit dextrin enzyme activity was measured using the Megazyme K-PULLG6 Pullulanase / Limit-Dextrinase Assay kit.

[0125] Temperature (℃) Activity of limit dextrinase (U / mL) Relative activity of limit dextrinase (%) 324.2480 375.31 100 425.0 194474.7990 503.7270 553.155960 2.274 3651.1020

[0126] Referring to Table 14 above, the optimum temperature for the limit dextrinase enzyme of the YBK-W4 strain was 37°C, and it showed 20% activity even at 65°C.

[0127] pH limit dextrinase activity (U / mL)Relative limit dextrinase activity (%)3.02.12404.02.40455.03.78716.04.52857.05.311008.04.73899.01.1321

[0128] Referring to Table 15 above, the optimal pH of the limit dextrinase enzyme of the YBK-W4 strain was 7.0, and it showed 89% activity even at pH 8.0.

[0129] Salinity of NaCl (%) Activity of limiting dextrinase (U / mL) Relative activity of limiting dextrinase (%) 05.311003.04.78906.03.78719.01.192212.00.407

[0130] Referring to Table 16 above, the limit dextrinase enzyme of the YBK-W4 strain showed 41% enzyme activity even at a salinity of 9%.

[0131] 5. Dextran lump reduction rate experiment

[0132] Dextran lump formation experiments of the W-4 strain were conducted by mixing and culturing 800 g of food waste, 200 g of molasses, and 50 mL of the inoculum in a 1 kg food waste disposer. Large-scale experiments were conducted by mixing and culturing 800 kg of livestock waste, 200 L of molasses, and 50 L of the inoculum in a 1 ton waste organic waste disposer. The mixture was stirred at 50 rpm for 48 hours at 50°C to examine the presence of clumps and lumps.

[0133] Processor CapacityUsed MicroorganismsClump production (kg)Reduction rate (%)1 KgConventional microorganisms0.110W-40.02901 TonConventional microorganisms5.150W-40.5390

[0134] Referring to Table 17 above, the dextran lump reduction rate of the YBK-W4 strain was 90% in the 1 kg treatment unit and the 1 ton treatment unit.

[0135] 6. Base sequence

[0136] Nucleotide sequence analysis was performed by Solgent on the 16s rRNA sequence. Homology comparison using the NCBI BLAST program revealed 99.6% homology with Bacillus atrophaeus strain CNY01. The 16s rRNA sequence is shown in Figure 7.

[0137] <Experiment on organic waste treatment using strains>

[0138] Bacillus atrophaeus YBK-W4 (KCTC 15572BP) strain was cultured in an incubator at an internal temperature of 50°C for 48 hours. The carrier contained 60% by volume of sawdust and 40% by volume of bamboo based on the total volume. 10% of the strain was adsorbed onto the carrier. 50% by volume of water based on the total volume was additionally mixed into the carrier and the culture medium, and the adsorption was performed while stirring and culturing for 48 hours at an internal temperature of 50°C. An organic waste treatment experiment was performed with the strain at a volume ratio of 1:1 between organic waste and the carrier on which the strain was adsorbed (organic waste: carrier on which the strain was adsorbed).

[0139] When the stirrer rotated inside the reaction vessel, a number of bamboo pieces were attached to the stirring blades of the stirrer and rotated together to help with stirring and quickly secure air gaps. The sizes of the bamboo pieces were 5 to 10 cm in width and 10 to 15 cm in length. Air was supplied into the inside of the reaction vessel and water vapor inside the reaction vessel was discharged. The dried waste air was discharged through a deodorizer.

[0140] [Experimental Example]

[0141] An organic waste treatment device using the above strain was installed at the Bukyung Pig Farming Cooperative's Bukyung Livestock Products Auction Center (Address: 23-100, Seobu-ro 1403beongil, Juchon-myeon, Gimhae-si, Gyeongsangnam-do; hereinafter referred to as "Bukyung Pig Farming") and the following experiments were conducted.

[0142] Experimental Example 1. Treatment of 5 tons of sludge

[0143] Five tons of sludge (waste) collected from the Bukyung pig slaughterhouse were fed into an organic waste treatment device using the above strain, and after 22 hours, clear condensate was discharged as a result of sludge treatment.

[0144] Experimental Example 2. Treatment of 2,500 kg of sludge cake and 2,500 kg of sludge solution.

[0145] 2,500 kg of sludge cake and 2,500 kg of sludge collected from the Bukyung Pig Slaughterhouse were fed into an organic waste treatment device using the above strain, and after 23 hours, clear condensate was discharged as a result of sludge and sludge treatment.

[0146] Experimental Example 3. Processing of 5 tons of slaughter by-products

[0147] Five tons of slaughterhouse by-products collected from the Bukyung Pig Slaughterhouse were put into an organic waste treatment device using the above strain. After 22 hours, condensate was discharged as a result of the slaughterhouse by-product treatment.

[0148] Experimental Example 4. Treatment of 5 tons of sludge

[0149] Five tons of sludge collected from the Bukyung Pig Slaughterhouse were fed into an organic waste treatment device using the above strain, and after 23 hours and 30 minutes, condensate was discharged as a result of sludge treatment.

[0150] Experimental Example 5. Treatment of 2,500 kg of slaughter by-products + 2,500 kg of sludge

[0151] 2,500 kg of slaughter by-products and 2,500 kg of sludge collected from the Bukyung Pig Slaughterhouse were fed into an organic waste treatment device using the above strain, and after 24 hours, clear condensate was discharged as a result of the slaughter by-products and sludge treatment.

[0152] The above description of the present invention is for illustrative purposes only, and those skilled in the art will readily appreciate that the present invention can be readily modified into other specific forms without altering the technical spirit or essential characteristics of the present invention. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive. For example, each component described as a single entity may be implemented in a distributed manner, and similarly, components described as distributed may be implemented in a combined manner.

[0153] The scope of the present invention is indicated by the claims described below rather than the detailed description above, and all changes or modifications derived from the meaning and scope of the claims and their equivalent concepts should be interpreted as being included in the scope of the present invention.

[0154] [Accession number]

[0155] Name of depositor: Korea Research Institute of Bioscience and Biotechnology, Biological Resource Center (KCTC)

[0156] Accession number: KCTC15572BP

[0157] Date of acceptance: 20230829

[0158]

Claims

1. Characterized by decomposing organic waste, Bacillus atrophaeus YBK-W4 (KCTC 15572BP) strain.

2. Obtaining a culture solution by culturing the strain according to paragraph 1; and A method for treating organic waste, comprising mixing the above-mentioned culture solution and organic waste to convert the organic waste into water.

3. In paragraph 2, A method for treating organic waste, wherein the above culturing is performed at a temperature range of 20°C to 80°C.

4. In paragraph 2, A method for treating organic waste, wherein the above culturing is performed for 24 to 72 hours.

5. In paragraph 2, A method for treating organic waste, wherein the above culture solution is used by being absorbed into a carrier.

6. In paragraph 5, A method for treating organic waste, wherein the carrier comprises sawdust and bamboo pieces.

7. In paragraph 6, A method for treating organic waste, wherein the sawdust is included in an amount of 10% to 80% by volume of the carrier.

8. In paragraph 6, A method for treating organic waste, wherein the bamboo pieces are included in an amount of 20% to 70% by volume of the carrier.

9. In paragraph 6, A method for treating organic waste, wherein the size of the above bamboo pieces is 1 cm to 50 cm in width and 5 cm to 40 cm in length.

10. In paragraph 5, A method for treating organic waste, wherein the above adsorption is performed at a temperature range of 20°C to 80°C.

11. In paragraph 5, A method for treating organic waste, wherein the above adsorption is performed for 12 to 72 hours.

12. In paragraph 2, A method for treating organic waste, wherein converting the organic waste into water is performed at a temperature range of 20°C to 80°C.

13. Using the strain according to Article 1, Organic waste treatment system.

14. In paragraph 13, The above organic waste treatment system includes an incubator and a reaction vessel, The strain is cultured in a liquid incubator to obtain a culture solution, An organic waste treatment system in which the culture medium and organic waste are mixed in the above reaction vessel and the organic waste is converted into water.

15. In paragraph 14, The above reaction vessel includes a stirring blade, Mixing the culture solution and the organic waste with the above stirring blade, Organic waste treatment system.

16. In paragraph 13, An organic waste treatment system, comprising an additional deodorizing system.

17. In paragraph 16, An organic waste treatment system, wherein the deodorizing system comprises spraying a strain according to claim 1 into the vaporized water from which the organic waste is converted.

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