KR20240015358A - Novel Bacillus licheniformis CD01 strain, composition for promotion using the same and manufacturing method thereof - Google Patents

Abstract

The present invention relates to a novel Bacillus licheniformis CD01 strain, a composition for promoting decay using the same, and a method for producing the same. Specifically, it relates to a novel Bacillus licheniformis CD01 strain (Accession number: KFCC) having decay promoting activity. 11936P), a composition for promoting decay containing the strain or its culture as an active ingredient, a decay accelerator for livestock manure or compost containing the composition as an active ingredient, and the step of spraying or mixing the composition into livestock manure or compost. It relates to a method of promoting the decay of livestock manure or compost and a method of producing a composition for promoting decay comprising the step of inoculating the strain or its culture into an organic fermentation source and fermenting it.

Description

Novel Bacillus licheniformis CD01 strain, composition for promoting decay using the same and manufacturing method thereof {Novel Bacillus licheniformis CD01 strain, composition for promotion using the same and manufacturing method thereof}

The present invention relates to a novel Bacillus licheniformis CD01 strain having cellulase and xylanase activities, a composition for promoting decay using the same, and a method for producing the same.

In 1970, with national economic development, meat consumption increased significantly due to the increase in national income level, and later, the number of livestock farms increased due to the Ministry of Environment's policy to encourage livestock farming, and as a result, the problem of handling livestock waste was also growing. According to the Ministry of Environment, livestock waste discharged from livestock raised in 150,000 livestock farms across the country amounts to 134,100 tons per day. Livestock manure that has not been properly composted and composted has a very high pollutant load (BOD, T-N, T-P) even when a small amount is spilled, and has a significant negative impact on groundwater and soil.

As a result, the Ministry of Environment implemented 'mandatory compost ripeness inspection' from March 25, 2020 in accordance with Article 15 of the Enforcement Rules of the Livestock Manure Act in order to improve problems such as bad odor and environmental pollution caused when livestock manure is spread on farmland. The guidance period was one year, and mandatory inspection took effect from March 25, 2021.

When livestock manure is composted by microorganisms, organic matter is converted into inorganic substances such as water, carbon dioxide, and ammonia, and other intermediate products and proliferated bacteria remain and are converted into humus. If you use a composting accelerator with appropriate aerobic microorganisms to effectively induce this composting process and achieve complete composting, you can reduce compost fermentation time and increase composting efficiency, thereby smoothing the operation of the composter, preventing water pollution, and using uncomposted compost. It can effectively reduce harmful gases such as ammonia gas that can affect the health of farmers.

Typical enzyme activities of microorganisms that can accelerate compost decomposition include xylanase and cellulase, and these enzyme activities include xylan and xylan, the main components of hemicellulose in livestock manure. It has the function of decomposing cellulose and promoting compost composting.

Meanwhile, the majority of compost composting agents currently on the market have both functions as supplementary feed and composting promotion, but their effectiveness is minimal. Accordingly, research has recently been conducted on the development of decay accelerators using microorganisms, but research on strains that have been identified as having excellent decay accelerator effects and have xylanase and cellulase activities, which are known to be effective in accelerating decay, is insufficient. This is the situation.

Accordingly, the present inventors isolated and identified a novel Bacillus strain with excellent xylanase and cellulase activity and decay-promoting activity from livestock manure and compost. When the identified strain was used, it effectively produced livestock manure and compost. The present invention was completed by confirming that it was possible to induce decay.

1. Republic of Korea registered patent 10-2179646

Therefore, the purpose of the present invention is to provide a Bacillus licheniformis CD01 strain (accession number: KFCC 11936P) having decay promoting activity.

Another object of the present invention is to provide a composition for promoting decay, comprising the Bacillus licheniformis CD01 strain or a culture thereof of the present invention as an active ingredient.

Another object of the present invention is to provide a decay accelerator for livestock manure or compost, comprising the composition of the present invention as an active ingredient.

Another object of the present invention is to provide a method for promoting the decay of livestock manure or compost, comprising the step of spraying or mixing the composition of the present invention into livestock manure or compost.

Another object of the present invention is to provide a method for producing a composition for promoting rot, comprising the step of inoculating an organic fermentation source with Bacillus licheniformis CD01 strain or a culture thereof and fermenting it.

In order to achieve the object of the present invention, the present invention provides a Bacillus licheniformis CD01 strain (accession number: KFCC 11936P) having decay promoting activity.

In one embodiment of the present invention, the strain may have both xylanase and cellulase activities.

In one embodiment of the present invention, the strain may promote the decay of livestock manure or compost.

In addition, the present invention provides a composition for promoting decay, comprising the Bacillus licheniformis CD01 strain or a culture thereof of the present invention as an active ingredient.

In one embodiment of the present invention, the Bacillus licheniformis CD01 strain or its culture may be included in an amount of 0.01 to 100% by weight based on the total weight of the composition.

In one embodiment of the present invention, the composition may be prepared by inoculating a culture of Bacillus licheniformis CD01 strain into a fermentation source containing rice bran and then fermenting it.

In one embodiment of the present invention, the culture of the rice bran and Bacillus licheniformis CD01 strain may be inoculated into the rice bran at a weight ratio of 10:1 to 9:2.

In one embodiment of the present invention, the fermentation may be performed at a temperature of 30 to 40 ° C. for 1 to 3 days.

In one embodiment of the present invention, the composition may maintain the Bacillus licheniformis CD01 strain at 2.0x10 ⁹ cfu/g or more.

Additionally, the present invention provides a decay accelerator for livestock manure or compost, comprising the composition of the present invention as an active ingredient.

The present invention also provides a method of promoting the decay of livestock manure or compost, comprising the step of spraying or mixing the composition of the present invention into livestock manure or compost.

Furthermore, the present invention provides a method for producing a composition for promoting rot, comprising the step of inoculating the Bacillus licheniformis CD01 strain or its culture of the present invention into an organic fermentation source and fermenting it.

In one embodiment of the present invention, the organic fermentation source may be rice bran.

In one embodiment of the present invention, the fermentation may be performed at a temperature of 30 to 40 ° C. for 1 to 3 days.

The Bacillus licheniformis CD01 strain according to the present invention is a strain with excellent cellulase and xylanase enzyme activities that promote compost composting. The composition for promoting composting containing the strain of the present invention effectively promotes the composting of livestock manure or compost. By enabling the production of high-quality compost, it can not only reduce the burden on livestock farms from mandatory compost ripeness tests, etc., but also shorten the composting time, which has the effect of helping livestock farms run their compost farms smoothly.

Figure 1 shows the results of streaking to pure isolate microorganisms from samples collected from livestock farms and compost in each region in order to discover microorganisms with decay-promoting activity in one embodiment of the present invention.
Figure 2 shows the results of congo red staining on solid medium supplemented with CMC in order to search for strains with both xylanase and cellulase activities from a single, purely isolated colony (A), and xylan The results of congo red staining on the added solid medium are shown (B), and the results of analyzing the xylanase and cellulase activities of the Bacillus licheniformis CD01 strain, which was selected as the most active strain, are shown. (C).
Figure 3 shows the degree of growth of the strain by measuring the absorbance for each culture time after inoculating the CD01 strain on different media to explore the optimal type of medium for culturing the Bacillus licheniformis CD01 strain identified in the present invention. It shows the results.
Figure 4 shows the results of analyzing the growth degree of the strain by measuring absorbance while culturing the CD01 strain under different pH and temperature conditions in order to establish the optimal culture conditions for the Bacillus licheniformis CD01 strain identified in the present invention. It is shown.
Figure 5 shows the process of inoculating the Bacillus licheniformis CD01 strain of the present invention into sterilized rice bran, which is the source of fermentation, and producing the spoilage accelerator of the present invention through fermentation and drying processes.
Figure 6 shows the results of confirming the number of viable bacteria before inoculating the Bacillus licheniformis CD01 strain into the fermentation source during the manufacturing process of the decay accelerator of the present invention and the results of confirming the number of viable bacteria contained in the manufactured decay accelerator.
Figure 7 shows the results of confirming the number of viable bacteria by requesting the rot accelerator of the present invention to the Korea Testing & Research Institute (KTR).
Figure 8 shows the results of visually confirming the degree of decay for the group treated with compost and the group not treated with the compost accelerator of the present invention.

The present invention relates to a novel Bacillus licheniformis CD01 strain and its use. Specifically, the present invention relates to a Bacillus licheniformis CD01 strain (accession number: KFCC 11936P) having decay-promoting activity. It has a distinctive feature in providing it.

In order to discover new microorganisms that can promote compost decomposition, the present inventors collected samples from livestock sheds and compost from farms in each region and identified microorganisms with compost-promoting activity from these. In particular, the present inventors screened microorganisms with both xylanase and cellulase activities as microorganisms with decay-promoting activity.

As a result, a strain with both xylanase and cellulase activities was isolated and identified, and as a result of 16s RNA base sequence analysis, it was confirmed that it was a new strain that had not been previously studied, and Bacillus licheniformis It was found that it was a strain belonging to the genus licheniformis .

Accordingly, the present inventors named the isolated strain as Bacillus licheniformis CD01 strain, deposited it with the Korea Microorganism Conservation Center on July 20, 2022, and received accession number KFCC 11936P.

The present inventors found that the Bacillus licheniformis CD01 strain identified in the present invention has both excellent xylanase and cellulase activities and can be used to promote rotting, especially in livestock manure and It was found that it can be a useful biological resource for composting.

Xylanase and cellulase, which are exocrine enzymes secreted from microorganisms, induce the activity of promoting compost decay by decomposing non-decomposable substances and using them as carbon sources.

In particular, cellulase, a non-degradable carbohydrate, is also known to contribute to the promotion of composting of spring manure, and xylan, a major component of hemicellulose, can be decomposed into D-xylose by xylanase, so xylanase increases the usability of agricultural by-products. It is known to be an enzyme necessary for growth.

In this regard, the Bacillus licheniformis CD01 strain identified in the present invention has both xylanase and cellulase activities and can be usefully used to promote the decay of livestock manure or compost.

Therefore, the present invention can provide a composition for promoting decay, comprising the Bacillus licheniformis CD01 strain or its culture of the present invention as an active ingredient.

The term ‘decomposition’ refers to the degree to which raw materials for compost and liquid fertilizer exhibit a stable response to plants and soil through the compost and liquid fertilizer process, and is a barometer that indicates the quality of organic fertilizer. If poorly composted compost is spread on the soil, ammonia generated during the decomposition process will cause a foul odor to the surrounding area and cause damage such as inhibition of crop growth due to ammonia gas.

The composition for promoting decay according to the present invention is a composition that has the activity of promoting the decay reaction so that this decay process can proceed quickly and completely. The active ingredient of the composition is Bacillus licheniformis CD01 of the present invention. It may include the strain itself or a culture or fermentation product of the strain.

The culture of the above strain can be obtained by culturing in large quantities by a typical culture method of microorganisms of the genus Bacillus licheniformis. As a culture medium, a medium containing a carbon source, a nitrogen source, vitamins and minerals can be used, and the carbon source Starch, sucrose, citrate, maltose, glucose, mannitol, glycerol, or xylose can be used.

In one embodiment of the present invention, in order to establish the optimal culture conditions for the Bacillus licheniformis CD01 strain, the growth degree of the strain was analyzed using various culture media, including molasses medium, soybean flour medium, and enzyme extraction medium. Compared to the group using TSB medium, the group using TSB medium showed the highest growth curve.

In addition, the optimal culture conditions were confirmed by culturing under different pH and temperature conditions. As a result, the strain was found to grow actively under conditions of pH 5 to 7, and in particular, the highest growth curve was observed at pH 6. , the culture temperature was found to be 30-40°C. Preferably, the optimal culture conditions for the strain use TSB medium, and can be cultured at pH 6 and a temperature of 30°C.

The culture can be used in the form of the culture itself containing the strain of the present invention, or the culture medium in the culture medium can be removed and only the concentrated cells can be recovered and used, and the concentrated cells can be frozen according to a conventional method ( It can be frozen or freeze-dried (lyophilized) to preserve its activity. Alternatively, the culture supernatant of the culture of the strain can also be used.

In one embodiment of the present invention, it was used in the form of the culture itself containing the strain.

Additionally, the Bacillus licheniformis CD01 strain or its culture may be included in an amount of 0.01 to 100% by weight based on the total weight of the composition.

In addition, the composition for promoting decay according to the present invention can be obtained through the step of inoculating the Bacillus licheniformis CD01 strain of the present invention into an organic fermentation source and fermenting it.

The strain itself or a culture of the strain may be used.

When using the fermentation source and the culture of the Bacillus licheniformis CD01 strain, the fermentation source and the culture of the strain can be mixed and fermented at a weight ratio of 10:1 to 9:2. In one embodiment of the present invention, Sterilized rice bran was used as a fermentation source, and sterilized rice bran and Bacillus licheniformis CD01 strain culture were inoculated at a weight ratio of 10:1, mixed, and fermented.

The fermentation can be performed at a temperature of 30 to 40°C for 1 to 3 days, and in one embodiment of the present invention, solid fermentation was performed at 37°C for 48 hours.

Furthermore, the present inventors measured the number of viable bacteria for the Bacillus licheniformis CD01 strain of the present invention contained in the composition for promoting decay of the present invention, and as a result, a high number of viable cells of 2.0x10 ⁹ cfu/g or more was contained in the composition for promoting decay. It appeared that it was done.

Through this, the composition for promoting rot (corrosion accelerator) manufactured using the strain of the present invention contains a large number of bacteria while maintaining the activity of the strain of the present invention, and the strain survives for a long time even during the storage period, showing that it has excellent preservability. could know that

Therefore, the present invention can provide a decay accelerator for livestock manure or compost, comprising the composition for promoting decay of the present invention.

The decay accelerator may be a microbial preparation containing the Bacillus licheniformis CD01 strain of the present invention, and may be in the form of a solid or liquid.

In the case of the solid form of the preparation, the microorganisms can be pulverized and commercialized in powder form, and in the case of using the microbial preparation in solid form, the manufacturing process is simple, the product production cost is low, and the preservability is good.

In one embodiment of the present invention, a rot accelerator in solid form was prepared.

In addition, when the decay accelerator of the present invention is used, the decay of livestock manure or compost can be further accelerated, and this acceleration of decay can also reduce odor generated from livestock manure.

The odor may be generated from odor gas derived from livestock manure, for example, from odor gas caused by anaerobic fermentation during the treatment of livestock manure.

Furthermore, the present invention can provide a method of promoting the decay of livestock manure or compost, comprising the step of spraying or mixing the composition for promoting decay of the present invention into livestock manure or compost.

In one embodiment of the present invention, the degree of composting was observed in the group that was not treated with the compost accelerator of the present invention and the group that was treated with the compost accelerator of the present invention. In the group treated with the compost accelerator of the present invention, the compost rotted quickly and well. It was found that progress was being made and that almost no foul odor was generated due to excrement. On the other hand, in the group that was not treated with the composting accelerator of the present invention, composting did not progress smoothly, so there was little change in the color or shape of the compost, and the foul odor of manure still appeared.

Through these results, the present inventors found that Bacillus licheniformis CD01 strain, a new strain isolated and identified in the present invention, can be usefully used as a microbial agent to promote the decay of livestock manure or compost.

Hereinafter, the components and technical features of the present invention will be described in more detail through the following examples. However, the following examples only illustrate the content of the present invention and the scope of the invention is not limited by the examples.

<Example 1>

Isolation and identification of strains with decay-promoting activity

<1-1> Sample collection and microorganism isolation from livestock and compost

To discover new microorganisms with decay-promoting activity, we visited Korean beef farms, pig farms, and chicken farms in each region, collected samples from livestock farms and fermenting compost, and performed a smear process for microbial screening. For this purpose, YM, TSA, PCA, PDA, and NA media, which are known in the art, were used, and the sampled samples were diluted with 1xPBS to prepare dilutions (10 ^-4 , 10 ^-5 , 10 ^-6 ). , the above diluted solution (100ul) was spread on each medium, and then cultured at 55°C to initially isolate microorganisms.

<1-2> Secondary pure separation

For secondary pure separation from the microorganisms isolated in <1-1>, colonies with different phenotypes are visually selected from the plate on which each microorganism is smeared, and streaking is performed to pure isolate the microorganisms from the colonies. was performed, and through streaking analysis, a purely isolated strain in the form of a single colony was isolated (see Figure 1).

<1-3> Isolation of strains with xylanase and cellulase activities

In order to isolate strains with both xylanase and cellulase activities from the single colony isolated in <1-2> above, it was confirmed on a medium to which CMC (carboxy methyl cellulose) or xylan were added as substrates, respectively. Cellulase active strains were isolated by inoculating the isolated strains on solid media supplemented with CMC and then measuring the size of the clear zone formed around the colonies. At this time, staining with 0.1% congo red solution and destaining with 1M NaCl were performed. Clear zone was checked. The congo red solution was prepared by adding 0.5 g of congo red powder to 500 ml of distilled water, mixing it, and then filtering it through a 0.22 um filter. For the destaining solution, distilled water was added to 45.66 g of NaCl, so that the total volume was 1 L. After being allowed to become , the solution filtered through a 0.22um filter was used. 10 ml of 0.1% congo red solution was injected into the plate where each strain was cultured and stained for 1 hour. Then, 1 M NaCl was injected at 10 ml each and destaining was repeated twice for 40 minutes. Afterwards, the clear zone (cellulase decomposition ring) formed on each plate was confirmed and a strain with cellulase activity was selected (Figure 2A). In addition, for the isolation of strains with xylanase activity, the strains were inoculated on a solid medium supplemented with 0.5% xylan as a substrate, and then, after culturing, the clear zone (xylan) formed on each plate was dyed and destained in the same manner as above. A strain with xylanase activity was selected by checking the size of the lanase decomposition ring (Figure 2B).

<1-4> Molecular biological identification of isolated strains

Through the process of <1-3>, the strain (strain 4-1) with the best cellulase and xylanase activities was isolated, and DNA was isolated from the strain to investigate the molecular biological characteristics of the isolated strain, followed by 16s The base sequence analysis of RNA sequencing was performed by requesting Cosmogenetech.

As a result, the identified strain was found to be a new strain belonging to Bacillus licheniformis , and the 16s RNA base sequence of the identified strain is as follows.

Bacillus licheniformis CD01 16s RNA base sequence:

AGGGTGATCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCGCAATGGACGAAAGTCTGACGGAGCAACGCCGCGTGAGTGATGAAGGTTTTCGGATCGTAAAACTCTGTTGTTAGGGAAGAACATGTACCGTTCGAATAGGGCGGTACCTTGACGGTACCTAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTG TCCGGAATTATTGGGCGTAAAGCGCGCGCAGGCGGTTCTTTAAGTCTGATGTGAAAGCCCCCGGCTCAACCGGGGAGGGTCATTGGAAACTGGGGAACTTGAGTGCAGAAGAGGAGAGTGGAATTCCACGTGTAGCGGTGAAATGCGTAGAGATGTGGAGGAACACCAGTGGCGAAGGCGACTCTCTGGTCTGTAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGAT GAGTGCTAAGTGTTAGAGGGTTTCCGCCCTTTAGTGCTGCAGCAAACGCATTAAGCACTCCGCCTGGGGAGTACGGTCGCAAGACTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCCTCTGACAACCCTAGAGATAGGGCTTCCCCTTCGGGGGCAGAGTGACAGGTGTGTGCATGGTTGTCGTCAGCTCGTGTCGTGA GATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGATCTTAGTTGCCAGCATTCAGTTGGGCACTCTAAGGTGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCC

In addition, the present inventors deposited the above strain with the Korea Microorganism Conservation Center on July 20, 2022, received accession number KFCC 11936P, and named the strain as ‘Bacillus licheniformis CD01 (Bacillus licheniformis CD01).’

<Example 2>

Establishment of optimal culture conditions

<2-1> Select badge type

In order to establish the optimal culture conditions for cultivating the Bacillus licheniformis CD01 strain of the present invention isolated through Example 1, an experiment was first performed to select the optimal type of medium. For this purpose, 1/2TSB medium, 1% molasses medium, soybean flour medium (MMS medium; soybean flour 3.6g + glucose2,25g + NaCl 1.13g + K2HPO4 1.13g + MgSO4 0.45g/450mL) and Yeast Extract medium (Dextrose 6.75g + K2HPO4) 1.35g + KH2PO4 0.9g + YeastExt 1.35g) were used respectively, each medium contained 0.1% CMC, and the strain was cultured at 37°C with stirring. Afterwards, 1 ml of strain culture was collected according to the incubation time, and the absorbance was measured at 600 nm to analyze the growth degree of the strain.

As a result, as shown in Table 1 and Figure 3, the group cultured in 1/2TSB medium showed significantly superior growth of the Bacillus licheniformis CD01 strain compared to the group cultured in other types of media. These results showed that using 1/2TSB medium is most suitable for cultivating Bacillus licheniformis CD01 strain.

<2-2> Selection of culture temperature and pH conditions

Next, to confirm the optimal culture temperature and pH conditions for culturing Bacillus licheniformis CD01 strain, prepare 1/2TSB medium with different pH conditions (pH 3 to 9) and inoculate the medium with CD01 strain. , culture was stirred at a temperature of 37°C, samples were collected over time, and absorbance was measured at 600 nm. At this time, 5.0N NaOH and 1N Citric acid were used to adjust pH.

Additionally, to confirm the optimal culture temperature, the CD01 strain was inoculated into 1/2TSB medium and cultured under different temperature conditions (20, 25, 30, 35, 40, 45, 50°C), and samples were collected over time. Absorbance was measured at 600 nm.

As a result, as shown in Figure 4, the growth of the Bacillus licheniformis CD01 strain was found to be most active under conditions of pH 5 to 7, and in particular, the highest absorbance value was shown at pH 6. Additionally, the optimal culture temperature conditions were found to be in the range of 30 to 40°C.

Through this, the present inventors found that for optimal cultivation of the Bacillus licheniformis CD01 strain isolated and identified in the present invention, 1/2TSB medium was used and cultured at pH 6 and a temperature of 30°C.

<Example 3>

Preparation of a composition for promoting decay using the strain of the present invention

The present inventors prepared a composition for promoting decay using the Bacillus licheniformis CD01 strain of the present invention by the following method. First, 1 L of the Bacillus licheniformis CD01 strain culture cultured under the optimal culture conditions identified in Example <2-1> was inoculated into 10 kg of sterilized (121°C, sterilized for 15 minutes) rice bran and incubated at 37°C for 48 hours. During solid fermentation. Afterwards, it was dried and then homogenized to prepare a decay accelerator using the Bacillus licheniformis CD01 strain of the present invention. A schematic diagram of the manufacturing process and product of the decay accelerator of the present invention described above is shown in Figure 5.

<Example 4>

Measurement of the number of bacteria contained in the composition for promoting decay of the present invention

In the process of manufacturing the composition for promoting rotting of Example 3, the sterilized rice bran powder of the present invention was added. The number of viable bacteria before inoculation with Bacillus licheniformis CD01 strain was analyzed, and the number of viable bacteria contained in the rot accelerator prepared through fermentation and drying process after inoculation was analyzed. In addition, the number of viable bacteria contained in the decay accelerator prepared in the present invention was analyzed by requesting the Korea Testing and Research Institute (KTR).

As a result, as shown in Figure 6, before inoculation, the viable cell count of the Bacillus licheniformis CD01 strain was found to be 4.8x10 ⁸ cfu/ml, but after inoculation, the viable cell count in the rot accelerator prepared through fermentation and drying process was It was found to be 2.2x10 ⁹ cfu/ml, and the KTR test results showed that it was 2.6x10 ⁹ cfu/ml (FIG. 7).

Through this, when the present inventors manufacture a composition for promoting rotting (corrosion accelerator) using the Bacillus licheniformis CD01 strain of the present invention, it is possible to manufacture a rotting accelerator that maintains a higher number of bacteria without reducing the number of bacteria during the manufacturing process. It was found that by maintaining a high bacterial count, improved decay-promoting activity can be shown, and preservability is also very high.

<Example 5>

Confirmation of the effect of promoting decay by the composition for promoting decay of the present invention

An experiment was performed to confirm the decay-promoting effect of the decay-promoting agent of the present invention prepared in Example 4 using the Bacillus licheniformis CD01 strain of the present invention. The degree of spoilage was visually confirmed for the group that was not treated with the compost accelerator of the present invention and the group that was treated with the compost collected from a Korean beef farm.

As a result, as shown in Figure 8, in the case of the group treated with the rotting accelerator of the present invention, compost composting was found to proceed quickly and well, and the odor of livestock manure was almost eliminated, showing that no foul odor was generated. On the other hand, in the group that was not treated with the composting accelerator of the present invention, composting did not proceed smoothly and there was little change in the color or shape of the compost, and the foul odor of livestock manure still appeared.

Through the above results, the present inventors have shown that the new strain identified in the present invention, Bacillus licheniformis CD01 strain, has both xylanase and cellulase activities and that the number of viable bacteria is increased without a decrease in the manufactured rot accelerator. It was found that the composting activity of compost or manure can be very effectively improved by indicating the number of viable bacteria.

So far, the present invention has been examined focusing on its preferred embodiments. A person skilled in the art to which the present invention pertains will understand that the present invention may be implemented in a modified form without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments should be considered from an illustrative rather than a restrictive perspective. The scope of the present invention is indicated in the claims rather than the foregoing description, and all differences within the equivalent scope should be construed as being included in the present invention.

Korea Center for Microbial Conservation (KCCM) KFCC11936P 20220720

Claims (14)

Bacillus licheniformis CD01 strain (Accession number: KFCC 11936P) with decay promoting activity. According to paragraph 1,
The strain is characterized by having both xylanase and cellulase activities. According to paragraph 1,
The strain is a strain characterized in that it promotes the decay of livestock manure or compost. A composition for promoting rot, comprising the Bacillus licheniformis CD01 strain of claim 1 or a culture thereof as an active ingredient. According to clause 4,
A composition for promoting decay, characterized in that the Bacillus licheniformis CD01 strain or culture thereof is contained in an amount of 0.01 to 100% by weight based on the total weight of the composition. According to clause 5,
The composition is a composition for promoting rotting, characterized in that it is manufactured by inoculating a culture of Bacillus licheniformis CD01 strain into a fermentation source containing rice bran and then fermenting it. According to clause 6,
A composition for promoting rotting, characterized in that the culture of the rice bran and the Bacillus licheniformis CD01 strain is inoculated into the rice bran at a weight ratio of 10:1 to 9:2. According to clause 6,
A composition for promoting rotting, characterized in that the fermentation is performed for 1 to 3 days at a temperature of 30 to 40 ° C. According to clause 4,
The composition is a composition for promoting rot, characterized in that the Bacillus licheniformis CD01 strain is maintained at 2.0x10 ⁹ cfu/g or more. A decay accelerator for livestock manure or compost, comprising the composition of claim 4 as an active ingredient. A method of promoting the decomposition of livestock manure or compost, comprising the step of spraying or mixing the composition of claim 4 into livestock manure or compost. A method for producing a composition for promoting rotting, comprising the step of inoculating the Bacillus licheniformis CD01 strain or a culture thereof of claim 1 into an organic fermentation source and fermenting it. According to clause 12,
A method for producing a composition for promoting rotting, wherein the organic fermentation source is rice bran. According to clause 12,
A method for producing a composition for promoting rotting, characterized in that the fermentation is performed at a temperature of 30 to 40 ° C. for 1 to 3 days.