WO2021112349A1 - Microbial preparation for decomposing organic wastes comprising novel geobacillus sp. strain and excipient - Google Patents

Microbial preparation for decomposing organic wastes comprising novel geobacillus sp. strain and excipient Download PDF

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WO2021112349A1
WO2021112349A1 PCT/KR2020/006370 KR2020006370W WO2021112349A1 WO 2021112349 A1 WO2021112349 A1 WO 2021112349A1 KR 2020006370 W KR2020006370 W KR 2020006370W WO 2021112349 A1 WO2021112349 A1 WO 2021112349A1
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strain
organic waste
geobacillus
present
waste
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PCT/KR2020/006370
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French (fr)
Korean (ko)
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전숭종
이주희
정유정
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동의대학교 산학협력단
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Priority claimed from KR1020190161850A external-priority patent/KR102175728B1/en
Priority claimed from KR1020190161856A external-priority patent/KR102283665B1/en
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Publication of WO2021112349A1 publication Critical patent/WO2021112349A1/en

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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
    • B09B—DISPOSAL OF SOLID WASTE NOT OTHERWISE PROVIDED FOR
    • B09B3/00—Destroying solid waste or transforming solid waste into something useful or harmless
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
    • B09B—DISPOSAL OF SOLID WASTE NOT OTHERWISE PROVIDED FOR
    • B09B5/00—Operations not covered by a single other subclass or by a single other group in this subclass
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
    • C05F17/20—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation using specific microorganisms or substances, e.g. enzymes, for activating or stimulating the treatment
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
    • Y02W30/00—Technologies for solid waste management
    • Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse

Definitions

  • the present invention relates to a microorganism preparation for decomposing organic waste comprising a novel strain of the genus Geobacillus and an excipient. More specifically, a method of decomposing organic waste using the decomposition activity of organic wastes of a novel Geobacillus sp. strain isolated from soil, etc., or providing a microbial agent capable of composting organic waste, decomposing or composting organic waste is to provide
  • Food waste (hereafter referred to as food waste) generated daily in Korea is 14,000 tons (Ministry of Environment, as of 2010), accounting for 28.7% of the total amount of waste generated every year. There is a trend of increasing by about 3% (Ministry of Environment, 2014). Food waste is causing enormous economic loss. It not only causes 800 billion won in treatment costs every year, 20 trillion won in economic loss (food resource value) per year, but also greenhouse gas emissions (8.85 million tons of CO2e per year), odor, and food. It is causing major environmental pollution such as water pollution due to wastewater (5.8% of the weight of food waste).
  • Microorganisms currently used for the treatment of organic waste such as food waste in Korea include Actinomycetes, Pseudomonas, Penicillium, Rhizopus, and Pichia.
  • the material decomposition ability is not excellent, so there is still a limit that treatment efficiency is reduced and odors may be generated.
  • a method of using mixed microorganisms including various microorganisms has been proposed as a method to complement the disadvantages of each microorganism, but this is because the conditions under which each microorganism is cultured, for example, pH and temperature, are different from each other.
  • the present invention provides a novel microorganism different from those of prior arts 1 and 2, and has a difference in the effect provided and its activity according to the novel microorganism.
  • Another object of the present invention is to provide a method for treating organic waste using the microbial agent.
  • Another object of the present invention is to provide a method for composting organic waste using the microbial agent.
  • the microorganism preparation for decomposing organic waste is to include a Geobacillus genus JH-8 (Geobacillus sp. JH-8) strain (accession number KCTC13853BP) and an excipient.
  • a Geobacillus genus JH-8 (Geobacillus sp. JH-8) strain (accession number KCTC13853BP) and an excipient.
  • the strain may have at least one enzyme activity selected from the group consisting of amylase, protease, and lipase.
  • the strain may be salt-tolerant.
  • the strain may have an optimum growth temperature of 30 to 80° C. and an optimum growth pH of 5 to 9.
  • the organic waste may be food waste or manure.
  • the manure is defined as including human or animal excrement.
  • the microorganism preparation for composting of organic waste is to include a Geobacillus genus JH-8 (Geobacillus sp. JH-8) strain (Accession No. KCTC13853BP), an excipient and a plant mixture.
  • a Geobacillus genus JH-8 Geobacillus sp. JH-8 strain (Accession No. KCTC13853BP)
  • an excipient and a plant mixture.
  • the organic waste may be food waste or manure.
  • the method for composting organic waste according to another embodiment of the present invention may include treating the microbial agent to the organic waste.
  • the method of treating organic waste according to another embodiment of the present invention may include treating the microbial agent to the organic waste.
  • the microorganism preparation for decomposing organic waste is to include a Geobacillus genus JH-8 (Geobacillus sp. JH-8) strain (accession number KCTC13853BP), an excipient and a plant mixture.
  • a Geobacillus genus JH-8 (Geobacillus sp. JH-8) strain (accession number KCTC13853BP)
  • an excipient accession number KCTC13853BP
  • the strain of the present invention is a novel strain of the genus Geobacillus JH-8, accession number KCTC13853BP.
  • the Geobacillus genus JH-8 (Geobacillus sp. JH-8) strain (Accession No.: KCTC13853BP) of the present invention may include 16s rRNA represented by SEQ ID NO: 1.
  • the novel strain of the present invention can grow well even at a high temperature of 50 ° C or higher among samples collected from soil around Gyeongsangnam-do, and has an enzyme capable of decomposing all of proteins, carbohydrates and fats, and at the same time in the process of decomposing organic waste.
  • a Gram-positive bacillus having the characteristic that it does not contain a malodor generating enzyme that may be generated, it was identified as a Geobacillus sp. strain through phylogenetic analysis using the nucleotide sequence of 16s rRNA represented by SEQ ID NO: 1.
  • the decomposition of organic waste of the present invention is a pretreatment that not only completely decomposes organic waste itself, but also facilitates incineration or landfill by hydrolyzing the binding of specific components of organic waste, for example, protein, fat and carbohydrate This means that all processes are included.
  • the organic waste of the present invention is, for example, food waste, sludge from sewage wastewater treatment plant, waste from aquatic products processing plant, waste from agricultural and livestock processing plant, waste from food paper mill, slaughter waste from slaughterhouse, livestock manure and other organic materials If it is a waste that includes, it may be included without being limited thereto.
  • the organic waste of the present invention may be food waste or manure, but is not limited thereto.
  • the manure of the present invention may be human or livestock manure, but is not limited thereto.
  • the microorganism preparation of the present invention may be in the form of a solid or liquid.
  • the moisture content is dried to 0.1 to 10% by weight and commercialized in the form of beads, or it can be pulverized and commercialized in powder form.
  • the Geobacillus genus JH-8 strain (accession number KCTC13853BP) is inoculated into the medium and cultured, and the obtained culture and the carrier are mixed and then dried through the process of drying the obtained microorganism fixation and microorganism It can be manufactured through the process of mixing nutrients, which are food ingredients of
  • any medium can be used as long as it is a medium capable of growth of the strain, and microbial nutrients commonly included in water, for example, yeast extract, glucose, sodium chloride, bovine extract, etc. may be used. .
  • the culture time of the strain of the present invention may be 12 hours to 48 hours, but is not limited thereto.
  • any known carrier for preparing a microbial agent may be used, for example, a vegetable carrier such as sawdust, rice husk, bran or powdery clay, activated carbon, coke, volcanic ash and combustion ash. At least one selected from the group may be used, and as the clay, at least one selected from the group consisting of zeolite, vermiculite, diatomaceous earth, kaolin, porcelain, feldspar, chard and talc may be used, but is not limited thereto. .
  • the adsorption property of the microbial agent can be increased, and it has the advantage that it can have a buffering ability for pH and can exhibit an adsorption effect on odor substances.
  • Various nutrients such as glucose, molasses, soy flour, sesame cake, fish meal, etc. may be used as the food component nutrients used in the microbial preparation of the present invention.
  • the nutrients may be mixed with the microbial fixation in a volume ratio of 0.2 to 1:10, but the present invention is not limited thereto.
  • the Geobacillus genus JH-8 strain (Accession No. KCTC13853BP) of the present invention has the advantage that it is not limited in nutrients because it can grow by decomposing various nutrients.
  • the microbial preparation of the present invention may further include an excipient.
  • the excipient of the present invention may be at least one selected from the group consisting of illite, activated carbon and elvan, but is not limited thereto. More preferably, the excipient may be included in a weight ratio of 0.1 to 5 times with respect to the microbial fixation, but is not limited thereto.
  • the microbial preparation of the present invention may further include a cryoprotectant.
  • the cryoprotectant may be glycerol, skim milk, honey, or the like, but is not limited thereto.
  • the microorganism preparation of the present invention is liquid, it may be used by mixing the culture of the microorganism and mixing glucose or glycerin to stabilize the microorganism, but is not limited thereto.
  • the microbial preparation of the present invention further comprises a plant mixture preparation, and the plant mixture preparation is Schisandra extract; calamus extract; eggplant leaf extract; dandelion extract; It may include any one selected from the group consisting of juniper extract and mixtures thereof.
  • the activity of the microorganisms is enhanced, so that the decomposition activity for organic wastes such as food waste and manure including human and animal excrement is increased, and the effect of removing odors can be increased.
  • the plant mixture includes Schisandra extract, 1 to 10 parts by weight of iris extract, 1 to 10 parts by weight of eggplant leaf extract, 0.5 to 5 parts by weight of dandelion extract, and 0.1 parts by weight of juniper extract based on 100 parts by weight of Schisandra extract. to 2 parts by weight.
  • the effect of promoting the activity of the microorganism is the most excellent, and thus the decomposition activity of organic waste and the effect of rapidly removing the odor may be the best.
  • the strain may have at least one enzymatic activity selected from the group consisting of amylase, protease and lipase, preferably amylase, protease and lipase. , but is not limited thereto.
  • the amylase of the present invention is a generic term for enzymes that hydrolyze starch, and is widely distributed in nature from higher plants to microorganisms, and hydrolyzes ⁇ -1,4-linkage or ⁇ -1,6-linkage of starch. has the function to Specifically, the amylase of the present invention is ⁇ -amylase ( ⁇ -1,4-glucan-4-glucano hydrolase), ⁇ -amylase ( ⁇ -1,4-glucan maltohydrolase) and ⁇ -amylase (amyllo glucosidase) may be at least one selected from the group consisting of, but is not limited thereto.
  • protease of the present invention is a generic term for enzymes that hydrolyze peptide bonds of proteins or peptides, and is also referred to as proteolytic enzymes.
  • the lipase of the present invention hydrolyzes triglycerides in a suspension of lipids and water to form monoglycerides, glycerol, and free fatty acids, as well as ester synthesis or exchange reaction from alcohols and fatty acids also catalyzes Since the lipase was first discovered in animal pancreas, it was found to be present in animal organs and plant seeds, and it was also found to be derived from microorganisms such as fungi and bacteria, confirming that it is widely distributed throughout the biological world. Of these, exocrine lipases produced by microorganisms have more heat stability than lipases derived from animals and plants, and thus can be applied for industrial purposes.
  • the strain may be salt-tolerant.
  • the strain of the present invention has the advantage of being able to decompose all proteins, carbohydrates, and fats while at the same time being resistant to salt, so that it can grow very effectively even in an environment in which food waste and manure containing a large amount of salt exist. .
  • the strain may have an optimum growth temperature of 30 to 80° C. and an optimum growth pH of 5 to 9.
  • the optimal growth temperature is 50 to 75 °C
  • the optimal growth pH may be 5.5 to 7
  • more preferably the optimal growth temperature is 70 °C
  • the optimal growth pH may be 6, but is not limited thereto. If the optimum growth temperature is less than the minimum value, the strain may not grow sufficiently, and if the optimum growth temperature exceeds the maximum value, the strain may be destroyed at high temperature. In addition, when it exceeds the minimum and maximum values of the optimal growth pH, the strain may be destroyed.
  • the strain of the present invention is composed of many organic substances such as carbohydrates, proteins, and fats, and the food waste, which accounts for most of the organic waste in Korea, has a pH of 4.0 to pH 6.5, so it very effectively decomposes the organic matter in the food waste. It can be applied very efficiently to the industrial fields of fermentation and extinction.
  • the present invention provides a culture of Geobacillus genus JH-8 (Geobacillus sp. JH-8) strain (Accession No. KCTC13853BP).
  • culture used in the present invention means a product obtained by culturing a microorganism in a known liquid medium or solid medium, and is a concept including microorganisms.
  • the culture is a product obtained after culturing the strain of the present invention in a medium
  • the medium may be selected from a known liquid medium or solid medium used for culturing the strain of the genus Geobacillus.
  • the present invention provides a culture solution from which the strain is removed from the culture of the genus Geobacillus JH-8 (Geobacillus sp. JH-8) (Accession No. KCTC13853BP).
  • culture solution refers to a product obtained by culturing microorganisms in a known liquid medium or solid medium, and is a concept that does not include microorganisms.
  • the culture is a product obtained after culturing the strain of the present invention in a medium
  • the medium may be selected from a known liquid medium or solid medium used for culturing the strain of the genus Geobacillus.
  • the organic waste may be food waste or manure.
  • the microorganism preparation for composting of organic waste is to include a Geobacillus genus JH-8 (Geobacillus sp. JH-8) strain (Accession No. KCTC13853BP), an excipient and a plant mixture.
  • a Geobacillus genus JH-8 Geobacillus sp. JH-8 strain (Accession No. KCTC13853BP)
  • an excipient and a plant mixture.
  • Composting of organic waste means composting which can fertilize the soil by hydrolyzing the bonds of certain components of the organic waste, such as proteins, fats and carbohydrates.
  • the organic waste may be food waste or manure.
  • the method for composting organic waste according to another embodiment of the present invention may include treating the microbial agent to the organic waste.
  • the information regarding the Geobacillus sp. JH-8 strain is the same as described in the strain and microbial preparation, so excessive complexity of the specification is reduced omitted to avoid
  • the method of treating organic waste according to another embodiment of the present invention may include treating the microbial agent to the organic waste.
  • the content regarding the Geobacillus sp. JH-8 strain (Accession No. KCTC13853BP) is the same as described in the strain and microbial preparation, and excessive complexity of the specification is reduced omitted to avoid
  • the novel Geobacillus genus JH-8 (Geobacillus sp. JH-8) strain of the present invention (accession number: KCTC13853BP) is capable of decomposing all proteins, carbohydrates and fats while at the same time has resistance to salt, so it contains a large amount of salt. It can grow very effectively even in food waste and manure that contain Furthermore, such a strain can reduce costs because the culture process is relatively simple, and there is an advantage of being environmentally friendly compared to the conventional chemical decomposition method.
  • 1 is a graph showing the optimum growth pH range of a novel strain according to an embodiment of the present invention.
  • Figure 2 is a graph showing the optimal growth temperature range of the novel strain according to an embodiment of the present invention.
  • FIG. 3 shows the results of genotyping using 16s rRNA of a novel strain according to an embodiment of the present invention.
  • the present invention relates to a microbial preparation for decomposing organic waste, including a Geobacillus genus JH-8 (Geobacillus sp. JH-8) strain (Accession No. KCTC13853BP), an excipient and a plant mixture.
  • a Geobacillus genus JH-8 Geobacillus sp. JH-8 strain (Accession No. KCTC13853BP)
  • an excipient and a plant mixture.
  • Strains that do not lose their activity even at 50° C. or higher were selected through the following procedure. Specifically, 1 g of various fungal sources samples collected from common soil, humus, and compost around Gyeongsangnam-do were suspended in 10 ml of 0.85% NaCl solution. Then, the suspension was diluted 10-fold, 100-fold, and 1000-fold to contain a nutrient medium (beef extract 3%, yeast extract 5%, agar 1.8%, pH 7.0). Plates were plated and incubated at 50 °C for 24 hours.
  • a nutrient medium beef extract 3%, yeast extract 5%, agar 1.8%, pH 7.0
  • Example 1 In order to confirm the growth of the strain, the strain of Example 1 was smeared on a plate containing a nutrient broth, incubated at 50 ° C. for 12 hours, and the growth rate according to the degree of colony formation was confirmed. The results are shown in Table 1 below.
  • the strain selected in Example 1 was inoculated on a plate containing a nutrient plate medium and cultured overnight at 50 °C. Then, iodine was treated on the plate in which the cultured strain is present, and the generation of a clear zone was checked, and the results are shown in Table 1 below.
  • 1% soluble starch in the case of amylase, 1% skim milk in the case of protease, and 1% tributyrin in the case of lipase were added to the nutritional plate medium.
  • Example 2 API kit (Biomerieux, USA) and morphology analysis were performed on the form of A-3 having all three enzyme activities, and the results are shown in Table 2 below.
  • the pH in the medium of Example 1 was adjusted to a range of 5.0 to 9.0, and the temperature was adjusted to a range of 30 to 80 ° C., and A-3 was cultured. Subsequently, the cell mass was measured, and the results are shown in FIGS. 2 and 3 .
  • A-3 was aerobic, and it was confirmed that the colony shape on the solid medium was round, the color was white, and the surface was in contact and smooth.
  • the A-3 is a gram-positive bacillus, has no flagella, formed endospores, did not produce H2S, urease, tryptophane deaminase, etc., and glucose, mannitol ( Various sugars such as mannitol) and sucrose were available.
  • the viable pH of A-3 was 5.0 to 9.0, and the temperature was 30 to 75 °C.
  • A-3 according to the present invention does not have an enzyme that can cause odor when decomposing organic matter, and can maintain growth by various nutrient sources, as well as organic waste, especially food waste. It can be seen that the organic waste treatment efficiency can be remarkably increased because it can be effectively propagated even when it contains a very high level of salt.
  • Chromosomes were extracted from the A-3 strain using the GENExTM genomic kit (General Biosystem, Korea). After mixing the extracted chromosome with Taq DNA polymerase (Takara shuzo, Japan) and the primers of SEQ ID NOs: 2 and 3, and performing Polymerase Chain Reaction to amplify 16s rRNA of strain A-3, , The nucleotide sequence was analyzed using an ABI PRISM 310 genetic analyzer (Perkin-Elmer Applied Biosystems, USA). In addition, in the case of homology analysis, the NCBI BLAST program was used, and the results are shown in FIG. 1 .
  • strain A-3 has the 16s rRNA sequence of SEQ ID NO: 1, and exhibits the highest 99% sequence homology with Geobacillus zalihae NBRC 101842.
  • A-3 was identified as a novel Geobacillus genus JH-8 (Geobacillus sp. JH-8) strain belonging to the genus Geobacillus. and deposited it with the Korea Research Institute of Bioscience and Biotechnology (Accession No.: KCTC13853BP).
  • a microbial preparation (JH8) for decomposing organic waste was prepared using the Geobacillus genus JH-8 (Geobacillus sp. JH-8) strain.
  • the time it takes for the microbial preparation to be treated with the following livestock manure (A) and food waste (B) and the odor is removed (decomposition rate 1, E1) and the time it takes for the weight to be reduced by half ( The degradation rate 2, E2) was evaluated.
  • Plant extracts as shown in [Table 4] below were prepared in order to evaluate the degree of enhancement of decomposition activity for organic wastes by mixing plant mixtures with respect to the microbial preparations.
  • Schisandra extract (S), iris extract (T), eggplant leaf extract (U), dandelion extract (X), and juniper extract (Z) were prepared by hot water extraction and filtration of each plant material.
  • the plant extract was mixed as shown in Table 4 below and mixed with the microbial preparation to prepare a microbial preparation for decomposing organic waste.
  • M1 M2 M3 M4 M5 M6 M7 M8 M9 S 100 100 100 100 100 100 T 100 0.1 One 5 10 15 U 100 0.1 One 5 10 15 X 100 0.1 0.5 3 5 7 Z 0.01 0.1 One 2 3
  • the prepared microbial preparation was treated in the manure (A), and the degradation rate 1 (E1) and degradation rate 2 (E2) were evaluated in the same manner as in the above-described method.
  • the index was fixed to 10 as a control when only a microbial preparation was used without using a plant mixture preparation, Comparative evaluation of examples in which the plant mixture preparation was mixed was evaluated as an index The results are shown. In the index, the lower the number, the better the effect.
  • the decomposition activity of microorganisms can be increased when the microorganisms are mixed with a plant extract of a certain range with respect to the preparation and used as a plant mixture preparation.
  • the present invention relates to a microorganism preparation for decomposing organic waste comprising a novel strain of the genus Geobacillus and an excipient. More specifically, a method of decomposing organic waste using the decomposition activity of organic wastes of a novel Geobacillus sp. strain isolated from soil, etc., or providing a microbial agent capable of composting organic waste, decomposing or composting organic waste is to provide

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Abstract

The present invention relates to a microbial preparation for decomposing organic wastes, comprising a novel Geobacillus sp. strain and an excipient and, more specifically, to a microbial preparation for decomposing organic wastes, comprising a novel Geobacillus sp. strain having a very high activity of decomposing organic wastes, a method for decomposing organic wastes using same, and a method for composting organic wastes. The strain can decompose all proteins, carbohydrates, and fats and, at the same time, is resistant to salt, and thus can very effectively grow even in food waste and excreta, which contain a large amount of salt. Moreover, the strain does not include an enzyme that generates bad odors when decomposing organic matter, and thus can be very suitable for use in decomposing organic wastes.

Description

신규한 지오바실러스속 균주 및 부형제를 포함하는 유기성 폐기물 분해용 미생물 제제Microbial preparation for decomposing organic waste containing novel Geobacillus strains and excipients
본 발명은 신규한 지오바실러스속 균주 및 부형제를 포함하는 유기성 폐기물 분해용 미생물 제제에 관한 것이다. 보다 상세하게는 토양 등에서 분리한 신규한 지오바실러스속 균주의 유기성 폐기물의 분해 활성을 이용하여 유기성 폐기물을 분해하거나, 유기성 폐기물을 퇴비화할 수 있는 미생물 제제를 제공하고, 유기성 폐기물을 분해하거나 퇴비화시키는 방법을 제공하기 위한 것이다.The present invention relates to a microorganism preparation for decomposing organic waste comprising a novel strain of the genus Geobacillus and an excipient. More specifically, a method of decomposing organic waste using the decomposition activity of organic wastes of a novel Geobacillus sp. strain isolated from soil, etc., or providing a microbial agent capable of composting organic waste, decomposing or composting organic waste is to provide
우리나라에서 매일 발생되는 음식물류 폐기물(이하 음폐물이라 한다)은 14,000여 톤(환경부, 2010년 기준)으로 전체 쓰레기 발생량의 28.7%를 차지하고 있으며, 음식문화, 생활수준의 향상, 식생활의 고급화 등으로 매년 3% 가량 증가하는 추세에 있다(환경부, 2014). 음폐물은 막대한 경제적 손실을 가져오고 있는데, 매년 처리비용 8천억 원, 연간 20조원 이상의 경제적 손실 (식량자원 가치)을 초래하고 있을 뿐 아니라 온실가스 배출(연간 885만 톤 CO2e), 악취 발생, 음식물폐수 (음폐물 중량의 5.8%)로 인한 수질오염 등 주요한 환경오염을 야기시키고있다.Food waste (hereafter referred to as food waste) generated daily in Korea is 14,000 tons (Ministry of Environment, as of 2010), accounting for 28.7% of the total amount of waste generated every year. There is a trend of increasing by about 3% (Ministry of Environment, 2014). Food waste is causing enormous economic loss. It not only causes 800 billion won in treatment costs every year, 20 trillion won in economic loss (food resource value) per year, but also greenhouse gas emissions (8.85 million tons of CO2e per year), odor, and food. It is causing major environmental pollution such as water pollution due to wastewater (5.8% of the weight of food waste).
현재 분리 수거되는 음폐물은 자원화(사료화 47.4%, 퇴비화 38.1%), 소각(소각물의 13.2%), 매립(매립물의 9.63%)으로 처리되고 있으나, 종량제의 합리적 운영을 위한 정책적 지원 및 관리체계의 부족, 사회·환경적 문제, 종량제 실시에 따른 각 배출원의 경제적 부담, 처리방법의 편리성에 대한 필요성으로 음폐물을 처리할 수Currently, separated and collected food waste is treated as resources (47.4% forage, 38.1% for compost), incineration (13.2% from incineration), and landfill (9.63% from landfill). Due to shortage, social/environmental problems, the economic burden of each emission source due to the implementation of the volume-rate system, and the necessity for the convenience of treatment methods,
있는 다양한 기술 개발 및 그 기술의 실생활 적용에 대한 수요가 점점 높아져 가고 있는 실정이다.There is a growing demand for the development of various technologies and the application of the technologies to real life.
이에 다양한 업체에서 가정용 음폐물 처리기를 개발하여 출시하고 있으며, 크게 열풍건조식, 분쇄건조식, 냉동보관방식, 가정용 음식물분쇄기 (디스포저) 방식, 미생물을 이용한 처리방식으로 분류된다. 그러나 미생물을 이용한 처리방식을 제외한 타 처리기술은 그 처리산물을 수거하여 2차 처리나 매립을 해야 하기 때문에 정부의 자원순환형 폐기물관리정책에 부응하지 못한다는 문제점을 갖는다. 따라서 음폐물 처리의 최우선 순위로 미생물을 이용한 처리방식에 의한 음폐물의 분해 또는 퇴비화하는 방안이 각광을 받고 있다.Accordingly, various companies have developed and released household waste treatment machines, and they are largely classified into hot air drying type, pulverizing drying type, freezing storage method, household food shredder (disposer) method, and treatment method using microorganisms. However, other treatment technologies, except for the treatment method using microorganisms, have a problem in that they cannot meet the government's resource circulation type waste management policy because the treated products must be collected and subjected to secondary treatment or landfill. Therefore, a method of decomposing or composting waste by a treatment method using microorganisms is in the spotlight as a top priority for waste treatment.
현재 국내에서 음식물 쓰레기와 같은 유기성 폐기물의 처리를 위해 이용되는 미생물로는 방선균류(Antinomycetes), 슈도모나스류(Pseudomonas), 페니실리움(Penicillium), 리조푸스(Rhizopus), 피치아(Pichia) 등이 있으나, 이들 미생물의 경우 물질 분해 능력이 우수하지 않아 처리 효율이 감소하고 악취가 발생될 수 있다는 한계점이 여전히 존재하는 실정이다. 또한, 미생물 각각의 단점을 상호보완하기 위한 하나의 방안으로 다양한 미생물을 포함하는 혼합 미생물 사용하는 방법이 제안되고 있으나, 이는 각 미생물이 배양되는 조건, 예를 들면 pH 및 온도 등이 서로 상이하기 때문에 미생물이 갖는 물질 분해 능력이 충분히 발휘될 수 없다는 한계점이 존재한다. 이런 한계점 극복을 위해, 단일 미생물을 사용한 경우에도 유기물 분해 효율이 충분히 발휘될 수 있는 새로운 균주의 활용이 필요한 실정이다. Microorganisms currently used for the treatment of organic waste such as food waste in Korea include Actinomycetes, Pseudomonas, Penicillium, Rhizopus, and Pichia. However, in the case of these microorganisms, the material decomposition ability is not excellent, so there is still a limit that treatment efficiency is reduced and odors may be generated. In addition, a method of using mixed microorganisms including various microorganisms has been proposed as a method to complement the disadvantages of each microorganism, but this is because the conditions under which each microorganism is cultured, for example, pH and temperature, are different from each other. There is a limit in that the ability of microorganisms to decompose substances cannot be sufficiently exhibited. In order to overcome this limitation, even when a single microorganism is used, it is necessary to utilize a new strain in which the decomposition efficiency of organic matter can be sufficiently exhibited.
관련하여, 선행기술 1(KR10-0424419 B1 (2004.03.12.)에는 '유기물 분해능이 우수한 신규 바실러스 에스피 균주 및 이를 이용한 음식물 쓰레기 처리방법'이 알려져 있으며, 선행기술 2(KR10-0443267 B1 (2004.07.23.)에는 폐유기물 분해활성이 우수한 신규 균주로서 바실러스리케니포미스 H3가 개시되어 있다.In relation to this, in the prior art 1 (KR10-0424419 B1 (2004.03.12.), 'a novel Bacillus sp. strain excellent in organic matter degradability and a food waste treatment method using the same' is known, and the prior art 2 (KR10-0443267 B1 (2004.07. 23.) discloses Bacillus licheniformis H3 as a novel strain with excellent decomposition activity of waste organic matter.
그러나 본 발명에서는 선행기술 1 및 2와 다른 신규한 미생물을 제공하는 것이며, 상기 신규한 미생물에 따라 제공되는 효과 및 그 활성에 차이를 가진다.However, the present invention provides a novel microorganism different from those of prior arts 1 and 2, and has a difference in the effect provided and its activity according to the novel microorganism.
본 발명의 목적은 지오바실러스속 JH-8(Geobacillus sp. JH-8) 균주(수탁번호 KCTC13853BP) 및 부형제를 포함하는 유기성 폐기물 분해용 및 유기성 폐기물의 퇴비화를 위한 미생물 제제를 제공하는 것이다.It is an object of the present invention to provide a microbial preparation for decomposing organic waste and composting of organic waste, including a Geobacillus sp. JH-8 strain (Accession No. KCTC13853BP) and excipients.
본 발명의 다른 목적은 상기 미생물 제제를 이용하여 유기성 폐기물을 처리하는 방법을 제공하는 것이다.Another object of the present invention is to provide a method for treating organic waste using the microbial agent.
본 발명의 다른 목적은 상기 미생물 제제를 이용하여 유기성 폐기물을 퇴비화하는 방법을 제공하는 제공하는 것이다.Another object of the present invention is to provide a method for composting organic waste using the microbial agent.
상기 목적을 달성하기 위하여, 본 발명의 일 실시예에 따른 유기성 폐기물 분해용 미생물 제제는 지오바실러스속 JH-8(Geobacillus sp. JH-8) 균주(수탁번호 KCTC13853BP) 및 부형제를 포함하는 것이다.In order to achieve the above object, the microorganism preparation for decomposing organic waste according to an embodiment of the present invention is to include a Geobacillus genus JH-8 (Geobacillus sp. JH-8) strain (accession number KCTC13853BP) and an excipient.
상기 균주는 아밀라제(amylase), 프로테아제(protease) 및 리파아제(lipase)로 구성된 군으로부터 선택되는 1종 이상의 효소 활성이 있는 것일 수 있다.The strain may have at least one enzyme activity selected from the group consisting of amylase, protease, and lipase.
상기 균주는 염분 내성이 있는 것일 수 있다.The strain may be salt-tolerant.
상기 균주는 최적 생장 온도가 30 내지 80 ℃이고, 최적 생장 pH가 5 내지 9 인 것일 수 있다.The strain may have an optimum growth temperature of 30 to 80° C. and an optimum growth pH of 5 to 9.
상기 유기성 폐기물 분해용 미생물 제제에 있어서, 유기성 폐기물은 음식물 쓰레기 또는 분뇨인 것일 수 있다. 본 발명에서 상기 분뇨는 인간 또는 동물의 배설물을 포함하는 것으로 정의한다.In the microbial preparation for decomposing organic waste, the organic waste may be food waste or manure. In the present invention, the manure is defined as including human or animal excrement.
본 발명의 다른 일 실시예에 따른 유기성 폐기물의 퇴비화를 위한 미생물 제제는 지오바실러스속 JH-8(Geobacillus sp. JH-8) 균주(수탁번호 KCTC13853BP), 부형제 및 식물 혼합제제를 포함하는 것이다.The microorganism preparation for composting of organic waste according to another embodiment of the present invention is to include a Geobacillus genus JH-8 (Geobacillus sp. JH-8) strain (Accession No. KCTC13853BP), an excipient and a plant mixture.
상기 유기성 폐기물의 퇴비화를 위한 미생물 제제에 있어서, 유기성 폐기물은 음식물 쓰레기 또는 분뇨인 것일 수 있다.In the microbial preparation for composting the organic waste, the organic waste may be food waste or manure.
본 발명의 다른 일 실시예에 따른 유기성 폐기물을 퇴비화하는 방법은 상기 미생물 제제를 유기성 폐기물에 처리하는 단계를 포함할 수 있다.The method for composting organic waste according to another embodiment of the present invention may include treating the microbial agent to the organic waste.
본 발명의 다른 일 실시예에 따른 유기성 폐기물을 처리하는 방법은 상기 미생물 제제를 유기성 폐기물에 처리하는 단계를 포함할 수 있다.The method of treating organic waste according to another embodiment of the present invention may include treating the microbial agent to the organic waste.
이하, 본 발명을 더욱 상세하게 설명한다.Hereinafter, the present invention will be described in more detail.
본 발명의 일 실시예에 따른 유기성 폐기물 분해용 미생물 제제는 지오바실러스속 JH-8(Geobacillus sp. JH-8) 균주(수탁번호 KCTC13853BP), 부형제 및 식물 혼합제제를 포함하는 것이다.The microorganism preparation for decomposing organic waste according to an embodiment of the present invention is to include a Geobacillus genus JH-8 (Geobacillus sp. JH-8) strain (accession number KCTC13853BP), an excipient and a plant mixture.
본 발명의 상기 균주는 지오바실러스 속 JH-8로서, 수탁번호 KCTC13853BP인 신규한 균주이다.The strain of the present invention is a novel strain of the genus Geobacillus JH-8, accession number KCTC13853BP.
본 발명의 상기 지오바실러스속 JH-8(Geobacillus sp. JH-8) 균주 (수탁번호: KCTC13853BP)는 서열번호 1로 표시되는 16s rRNA를 포함할 수 있다.The Geobacillus genus JH-8 (Geobacillus sp. JH-8) strain (Accession No.: KCTC13853BP) of the present invention may include 16s rRNA represented by SEQ ID NO: 1.
본 발명의 상기 신규한 균주는 경상남도 일대의 토양에서 채취한 시료 중에서, 50 ℃ 이상의 고온에서도 잘 성장할 수 있으며, 단백질, 탄수화물 및 지방 모두를 분해할 수 있는 효소를 가지고, 동시에 유기성 폐기물의 분해 과정에서 발생될 수 있는 악취 생성 효소를 포함하지 않는 특징을 가지는 그람 양성 간균으로서, 상기 서열번호 1로 표시되는 16s rRNA의 염기 서열을 이용한 계통 분석을 통해 지오바실러스 속 균주로 동정하였다.The novel strain of the present invention can grow well even at a high temperature of 50 ° C or higher among samples collected from soil around Gyeongsangnam-do, and has an enzyme capable of decomposing all of proteins, carbohydrates and fats, and at the same time in the process of decomposing organic waste. As a Gram-positive bacillus having the characteristic that it does not contain a malodor generating enzyme that may be generated, it was identified as a Geobacillus sp. strain through phylogenetic analysis using the nucleotide sequence of 16s rRNA represented by SEQ ID NO: 1.
본 발명의 유기성 폐기물의 분해란 유기성 폐기물의 특정 성분, 예를 들면 단백질, 지방 및 탄수화물의 결합을 가수분해 시킴으로써, 그 자체로써 유기성 폐기물을 완전히 분해하는 것 뿐만 아니라, 소각 또는 매립이 용이하도록 하는 전처리 과정을 모두 포함하는 것을 의미한다.The decomposition of organic waste of the present invention is a pretreatment that not only completely decomposes organic waste itself, but also facilitates incineration or landfill by hydrolyzing the binding of specific components of organic waste, for example, protein, fat and carbohydrate This means that all processes are included.
본 발명의 상기 유기성 폐기물은 예를 들면, 음식물 쓰레기, 하수도 폐수처리장의 슬러지, 수산물 가공공장의 폐기물, 농축산물 가공공장의 폐기물, 식품제지 공장의 찌꺼기, 도축장의 도축 폐기물, 축산분뇨 및 기타 유기성 물질을 포함하는 폐기물이라면 이에 제한되지 않고 모두 포함될 수 있다. 바람직하게 본 발명의 상기 유기성 폐기물은 음식물 쓰레기 또는 분뇨일 수 있으나, 이에 제한되는 것은 아니다.The organic waste of the present invention is, for example, food waste, sludge from sewage wastewater treatment plant, waste from aquatic products processing plant, waste from agricultural and livestock processing plant, waste from food paper mill, slaughter waste from slaughterhouse, livestock manure and other organic materials If it is a waste that includes, it may be included without being limited thereto. Preferably, the organic waste of the present invention may be food waste or manure, but is not limited thereto.
본 발명의 상기 분뇨는 인간 또는 가축의 분뇨일 수 있으나, 이에 제한되는 것은 아니다.The manure of the present invention may be human or livestock manure, but is not limited thereto.
본 발명의 상기 미생물 제제는 고체 또는 액상의 형태일 수 있다. 상기 고체 형태의 미생물제제의 경우는 상기 본 발명에 따른 상기 균주를 담체에 부착시킨 후 수분 함량이 0.1 내지 10 중량%가 되도록 건조시키고 비드 형태 제품화하거나, 분쇄하여 분말 형태로 제품화할 수 있다.The microorganism preparation of the present invention may be in the form of a solid or liquid. In the case of the microbial preparation in the solid form, after attaching the strain according to the present invention to the carrier, the moisture content is dried to 0.1 to 10% by weight and commercialized in the form of beads, or it can be pulverized and commercialized in powder form.
상기 미생물 제제가 고체 형태인 경우, 상기 지오바실러스속 JH-8 균주(수탁번호 KCTC13853BP)를 배지에 접종하여 배양하고, 수득된 배양물과 담체를 혼합한 뒤 건조하는 과정을 통해 얻어진 미생물 고정물과 미생물의 먹이 성분인 영양소를 혼합하는 과정을 통해 제조될 수 있다.When the microbial preparation is in a solid form, the Geobacillus genus JH-8 strain (accession number KCTC13853BP) is inoculated into the medium and cultured, and the obtained culture and the carrier are mixed and then dried through the process of drying the obtained microorganism fixation and microorganism It can be manufactured through the process of mixing nutrients, which are food ingredients of
본 발명의 상기 배지는 상기 균주의 생장이 가능한 배지라면 어떠한 것이라도 사용될 수 있고, 물에 통상적으로 포함되는 미생물 영양분, 예를 들면 효모 추출물, 글루코스, 염화나트륨, 소 추출물 등이 첨가된 것을 사용할 수 있다.As the medium of the present invention, any medium can be used as long as it is a medium capable of growth of the strain, and microbial nutrients commonly included in water, for example, yeast extract, glucose, sodium chloride, bovine extract, etc. may be used. .
본 발명의 상기 균주의 배양 시간은 12시간 내지 48시간일 수 있으나, 이에 제한되는 것은 아니다.The culture time of the strain of the present invention may be 12 hours to 48 hours, but is not limited thereto.
본 발명의 상기 담체는 미생물 제제를 제조하기 위한 공지의 담체를 모두 사용할 수 있고, 예를 들면 톱밥, 왕겨, 밀기울과 같은 식물성 담체 또는 분 말상의 점토류, 활성탄, 코크스, 화산재 및 연소재로 구성된 군으로부터 선택되는 적어도 하나가 사용될 수 있으며, 상기 점토류로는 제올라이트, 질석, 규조토, 고령토, 옹기토, 장석, 차지토 및 활석으로 구성된 군으로부터 선택되는 적어도 하나를 사용할 수 있으나 이에 제한되는 것은 아니다. 이와 같이 무기 담체를 이용하는 경우에는 미생물 제제의 흡착성을 높일 수 있을 뿐만 아니라, pH에 대한 완충 능력을 가질 수 있고 악취물질에 대한 흡착 효과를 발휘할 수 있다는 장점을 갖는다.As the carrier of the present invention, any known carrier for preparing a microbial agent may be used, for example, a vegetable carrier such as sawdust, rice husk, bran or powdery clay, activated carbon, coke, volcanic ash and combustion ash. At least one selected from the group may be used, and as the clay, at least one selected from the group consisting of zeolite, vermiculite, diatomaceous earth, kaolin, porcelain, feldspar, chard and talc may be used, but is not limited thereto. . In the case of using the inorganic carrier as described above, the adsorption property of the microbial agent can be increased, and it has the advantage that it can have a buffering ability for pH and can exhibit an adsorption effect on odor substances.
본 발명의 상기 미생물 제제에 사용되는 먹이 성분 영양소는 포도당, 당밀, 콩가루, 깻묵, 어분 등과 같은 다양한 영양소가 사용될 수 있다. 바람직하게 상기 영양소는 상기 미생물 고정물과 0.2~1:10의 부피비로 혼합될 수 있으나, 이에 제한되는 것은 아니다. 본 발명의 상기 지오바실러스속 JH-8 균주(수탁번호 KCTC13853BP)는 다양한 영양소를 분해하여 성장할 수 있기 때문에 영양소에 제한을 받지 않는다는 장점이 존재한다.Various nutrients such as glucose, molasses, soy flour, sesame cake, fish meal, etc. may be used as the food component nutrients used in the microbial preparation of the present invention. Preferably, the nutrients may be mixed with the microbial fixation in a volume ratio of 0.2 to 1:10, but the present invention is not limited thereto. The Geobacillus genus JH-8 strain (Accession No. KCTC13853BP) of the present invention has the advantage that it is not limited in nutrients because it can grow by decomposing various nutrients.
본 발명의 상기 미생물 제제는 부형제를 더 포함할 수 있다. 바람직하게 본 발명의 상기 부형제는 일라이트, 활성탄 및 맥반석으로 이루어진 군으로부터 선택되는 적어도 하나일 수 있으나, 이에 제한되는 것은 아니다. 더욱 바람직하게 상기 부형제는 상기 미생물 고정물에 대하여 0.1 내지 5배의 중량비로 포함될 수 있으나, 이에 제한되는 것은 아니다.The microbial preparation of the present invention may further include an excipient. Preferably, the excipient of the present invention may be at least one selected from the group consisting of illite, activated carbon and elvan, but is not limited thereto. More preferably, the excipient may be included in a weight ratio of 0.1 to 5 times with respect to the microbial fixation, but is not limited thereto.
본 발명의 상기 미생물 제제는 동결 보호제를 더 포함할 수 있다. 바람직하게 상기 동결 보호제는 글리세롤, 탈지유, 꿀 등이 사용될 수 있으나, 이에 제한되는 것은 아니다.The microbial preparation of the present invention may further include a cryoprotectant. Preferably, the cryoprotectant may be glycerol, skim milk, honey, or the like, but is not limited thereto.
본 발명의 상기 미생물 제제가 액상인 경우, 미생물의 배양물을 혼합하고 미생물을 안정화시키기 위해 포도당이나 글리세린을 혼합하여 사용할 수 있으나, 이에 제한되는 것은 아니다.When the microorganism preparation of the present invention is liquid, it may be used by mixing the culture of the microorganism and mixing glucose or glycerin to stabilize the microorganism, but is not limited thereto.
바람직하게 본 발명의 상기 미생물 제제는 식물 혼합제제를 더 포함하는 것이고, 상기 식물 혼합 제제는 오미자 추출물; 창포 추출물; 가지 잎 추출물; 민들레 추출물; 노간주나무 추출물 및 이들의 혼합물로 이루어진 군에서 선택된 어느 하나를 포함하는 것일 수 있다.Preferably, the microbial preparation of the present invention further comprises a plant mixture preparation, and the plant mixture preparation is Schisandra extract; calamus extract; eggplant leaf extract; dandelion extract; It may include any one selected from the group consisting of juniper extract and mixtures thereof.
상기 식물 혼합제제가 포함되는 경우 상기 미생물의 활성이 증진되어 식품 폐기물, 인간 및 동물의 배설물을 포함하는 분뇨 등 유기성 폐기물에 대한 분해 활성이 높아지고, 악취를 제거하는 효과가 높아질 수 있다.When the plant mixture is included, the activity of the microorganisms is enhanced, so that the decomposition activity for organic wastes such as food waste and manure including human and animal excrement is increased, and the effect of removing odors can be increased.
더 바람직하게 상기 식물 혼합제제는 오미자 추출물을 포함하고 상기 오미자 추출물 100 중량부에 대하여 창포 추출물 1 내지 10 중량부, 가지 잎 추출물 1 내지 10 중량부, 민들레 추출물 0.5 내지 5 중량부 및 노간주나무 추출물 0.1 내지 2 중량부를 포함하는 것일 수 있다.More preferably, the plant mixture includes Schisandra extract, 1 to 10 parts by weight of iris extract, 1 to 10 parts by weight of eggplant leaf extract, 0.5 to 5 parts by weight of dandelion extract, and 0.1 parts by weight of juniper extract based on 100 parts by weight of Schisandra extract. to 2 parts by weight.
상기 범위에 의하는 경우 상기 미생물의 활성을 증진하는 효과가 가장 우수하여 유기성 폐기물의 분해 활성 및 악취를 빠르게 제거하는 효과가 가장 우수할 수 있다.According to the above range, the effect of promoting the activity of the microorganism is the most excellent, and thus the decomposition activity of organic waste and the effect of rapidly removing the odor may be the best.
상기 균주는 아밀라제(amylase), 프로테아제(protease) 및 리파아제(lipase)로 구성된 군으로부터 선택되는 1종 이상의 효소 활성이 있는 것일 수 있고, 바람직하게는 아밀라제, 프로테아제 및 리파아제 효소 활성을 모두 가지는 것일 수 있으나, 이에 제한되는 것은 아니다.The strain may have at least one enzymatic activity selected from the group consisting of amylase, protease and lipase, preferably amylase, protease and lipase. , but is not limited thereto.
본 발명의 상기 아밀라제는 전분을 가수분해하는 효소의 총칭으로서, 고등식물부터 미생물에까지 자연계에 널리 분포하며, 녹말의 α-1,4-연결(linkage) 또는α-1,6-연결을 가수분해하는 기능을 갖는다. 구체적으로 본 발명의 상기 아밀라제는 α-아밀라제(α-1,4-글루칸-4-글루카노 하이드로라제), β-아밀라제(α-1,4-글루칸 말토 하이드로라제) 및 γ-아밀라제(아밀로글루코시다제)로 구성된 군으로부터 선택되는 적어도 하나일 수 있으나, 이에 제한되는 것은 아니다.The amylase of the present invention is a generic term for enzymes that hydrolyze starch, and is widely distributed in nature from higher plants to microorganisms, and hydrolyzes α-1,4-linkage or α-1,6-linkage of starch. has the function to Specifically, the amylase of the present invention is α-amylase (α-1,4-glucan-4-glucano hydrolase), β-amylase (α-1,4-glucan maltohydrolase) and γ-amylase (amyllo glucosidase) may be at least one selected from the group consisting of, but is not limited thereto.
본 발명의 상기 프로테아제는 단백질이나 펩티드의 펩타이드 결합을 가수분해하는 효소를 총칭하는 것으로서, 단백질 분해 효소라고도 일컫는다.The protease of the present invention is a generic term for enzymes that hydrolyze peptide bonds of proteins or peptides, and is also referred to as proteolytic enzymes.
본 발명의 상기 리파아제는 지질과 물의 현탁액 상태에서 트리글리세리드(triglycerides)를 가수분해하여 모노글리세리드(monoglycerides), 글리세롤(glycerol), 유리 지방산을 형성할 뿐만 아니라 알코올과 지방산으로부터 에스테르(ester) 합성 또는 교환 반응을 촉매하기도 한다. 상기 리파아제는 동물의 췌액에서 처음 발견된 이래, 동물의 기관, 식물의 종자 내에 존재함이 밝혀졌으며, 곰팡이, 세균 등의 미생물로부터 유래된 것도 발견되어 생물계 전반에 널리 분포되어 있음이 확인되었다. 이 중, 미생물이 생산하는 외분비형 리파아제는 동물과 식물 유래의 리파아제 보다 열에 더 안정한 특징이 존재하여 산업용으로 적용될 수 있다.The lipase of the present invention hydrolyzes triglycerides in a suspension of lipids and water to form monoglycerides, glycerol, and free fatty acids, as well as ester synthesis or exchange reaction from alcohols and fatty acids also catalyzes Since the lipase was first discovered in animal pancreas, it was found to be present in animal organs and plant seeds, and it was also found to be derived from microorganisms such as fungi and bacteria, confirming that it is widely distributed throughout the biological world. Of these, exocrine lipases produced by microorganisms have more heat stability than lipases derived from animals and plants, and thus can be applied for industrial purposes.
상기 균주는 염분 내성이 있는 것일 수 있다.The strain may be salt-tolerant.
염분 내성을 갖는 경우 매우 높은 염분을 갖는 음식물 쓰레기와 같은 유기성 폐기물이 존재하는 환경에서 효과적으로 생육될 수 있다.If it has salt tolerance, it can be effectively grown in an environment in which organic waste such as food waste having a very high salinity exists.
본 발명의 균주는 단백질, 탄수화물 및 지방 모두를 분해할 수 있으면서 동시에 염분에 대한 내성이 존재하기 때문에 염분이 다량으로 포함되어 있는 음식물 쓰레기 및 분뇨가 존재하는 환경에서도 매우 효과적으로 성장할 수 있다는 장점이 존재한다.The strain of the present invention has the advantage of being able to decompose all proteins, carbohydrates, and fats while at the same time being resistant to salt, so that it can grow very effectively even in an environment in which food waste and manure containing a large amount of salt exist. .
상기 균주는 최적 생장 온도가 30 내지 80 ℃이고, 최적 생장 pH가 5 내지 9 인 것일 수 있다.The strain may have an optimum growth temperature of 30 to 80° C. and an optimum growth pH of 5 to 9.
바람직하게는 최적 생장 온도가 50 내지 75 ℃이고, 최적 생장 pH가 5.5 내지 7일 수 있으며, 더욱 바람직하게 최적 생장 온도가 70 ℃이고, 최적 생장 pH가 6일 수 있으나, 이에 제한되는 것은 아니다. 상기 최적 생장 온도 최저 값 미만인 경우에는 균주가 충분히 성장할 수 없으며, 최적 성장 온도 최대 값을 초과하는 경우에는 고온에 균주가 파괴될 수 있다. 또한, 상기 최적 생장 pH의 최저값 미만 및 최대값을 초과하는 경우에는 균주가 파괴될 수 있다.Preferably, the optimal growth temperature is 50 to 75 °C, the optimal growth pH may be 5.5 to 7, more preferably the optimal growth temperature is 70 °C, and the optimal growth pH may be 6, but is not limited thereto. If the optimum growth temperature is less than the minimum value, the strain may not grow sufficiently, and if the optimum growth temperature exceeds the maximum value, the strain may be destroyed at high temperature. In addition, when it exceeds the minimum and maximum values of the optimal growth pH, the strain may be destroyed.
본 발명의 상기 균주는 대한민국의 유기성 폐기물의 대부분을 차지하는 음식물쓰레기는 탄수화물, 단백질, 지방 등 많은 유기물로 이루어져 있으며, pH가 pH 4.0 ~ pH 6.5에 해당하기 때문에, 음식물쓰레기 내의 유기물을 매우 효과적으로 분해하여 발효, 소멸화 하는 산업 분야에 매우 효율적으로 적용될 수 있다.The strain of the present invention is composed of many organic substances such as carbohydrates, proteins, and fats, and the food waste, which accounts for most of the organic waste in Korea, has a pH of 4.0 to pH 6.5, so it very effectively decomposes the organic matter in the food waste. It can be applied very efficiently to the industrial fields of fermentation and extinction.
본 발명은 지오바실러스속 JH-8(Geobacillus sp. JH-8) 균주(수탁번호 KCTC13853BP)의 배양물을 제공한다.The present invention provides a culture of Geobacillus genus JH-8 (Geobacillus sp. JH-8) strain (Accession No. KCTC13853BP).
본 발명에서 사용되는 용어 배양물이란 미생물을 공지의 액체 배지 또는 고체 배지에서 배양시켜 수득한 산물을 의미하여, 미생물이 포함되는 개념이다.The term culture used in the present invention means a product obtained by culturing a microorganism in a known liquid medium or solid medium, and is a concept including microorganisms.
본 발명에서 배양물은 본 발명의 균주를 배지에서 배양시킨 후 얻어진 산물로서, 상기 배지는 지오바실러스속 균주 배양에 사용되는 공지의 액체 배지 또는 고체 배지에서 선택될 수 있다.In the present invention, the culture is a product obtained after culturing the strain of the present invention in a medium, and the medium may be selected from a known liquid medium or solid medium used for culturing the strain of the genus Geobacillus.
또한, 본 발명은 지오바실러스속 JH-8(Geobacillus sp. JH-8) 균주(수탁번호 KCTC13853BP)의 배양물로부터 상기 균주가 제거된 배양액을 제공한다.In addition, the present invention provides a culture solution from which the strain is removed from the culture of the genus Geobacillus JH-8 (Geobacillus sp. JH-8) (Accession No. KCTC13853BP).
본 발명에서 사용되는 용어 "배양액"이란 미생물을 공지의 액체 배지 또는 고체 배지에서 배양시켜 수득한 산물을 의미하여, 미생물이 포함되지 않는 개념이다.As used herein, the term "culture solution" refers to a product obtained by culturing microorganisms in a known liquid medium or solid medium, and is a concept that does not include microorganisms.
본 발명에서 배양물은 본 발명의 균주를 배지에서 배양시킨 후 얻어진 산물로서, 상기 배지는 지오바실러스속 균주 배양에 사용되는 공지의 액체 배지 또는 고체 배지에서 선택될 수 있다.In the present invention, the culture is a product obtained after culturing the strain of the present invention in a medium, and the medium may be selected from a known liquid medium or solid medium used for culturing the strain of the genus Geobacillus.
상기 유기성 폐기물 분해용 미생물 제제에 있어서, 유기성 폐기물은 음식물 쓰레기 또는 분뇨인 것일 수 있다.In the microbial preparation for decomposing organic waste, the organic waste may be food waste or manure.
본 발명의 다른 일 실시예에 따른 유기성 폐기물의 퇴비화를 위한 미생물 제제는 지오바실러스속 JH-8(Geobacillus sp. JH-8) 균주(수탁번호 KCTC13853BP), 부형제 및 식물 혼합제제를 포함하는 것이다.The microorganism preparation for composting of organic waste according to another embodiment of the present invention is to include a Geobacillus genus JH-8 (Geobacillus sp. JH-8) strain (Accession No. KCTC13853BP), an excipient and a plant mixture.
유기성 폐기물의 퇴비화란 유기성 폐기물의 특정 성분, 예를 들면 단백질, 지방 및 탄수화물의 결합을 가수분해 시킴으로써 토양을 비옥하게 할 수 있는 퇴비화하는 것을 의미한다.Composting of organic waste means composting which can fertilize the soil by hydrolyzing the bonds of certain components of the organic waste, such as proteins, fats and carbohydrates.
상기 유기성 폐기물의 퇴비화를 위한 미생물 제제에 있어서, 유기성 폐기물은 음식물 쓰레기 또는 분뇨인 것일 수 있다.In the microbial preparation for composting the organic waste, the organic waste may be food waste or manure.
본 발명의 다른 일 실시예에 따른 유기성 폐기물을 퇴비화하는 방법은 상기 미생물 제제를 유기성 폐기물에 처리하는 단계를 포함할 수 있다.The method for composting organic waste according to another embodiment of the present invention may include treating the microbial agent to the organic waste.
본 발명의 처리하는 방법에서, 상기 지오바실러스속 JH-8(Geobacillus sp. JH-8) 균주(수탁번호 KCTC13853BP)에 관한 내용은 상기 균주 및 미생물 제제에서 기재한 바와 동일하여, 명세서의 과도한 복잡성을 피하기 위하여 생략한다.In the treatment method of the present invention, the information regarding the Geobacillus sp. JH-8 strain (Accession No. KCTC13853BP) is the same as described in the strain and microbial preparation, so excessive complexity of the specification is reduced omitted to avoid
본 발명의 다른 일 실시예에 따른 유기성 폐기물을 처리하는 방법은 상기 미생물 제제를 유기성 폐기물에 처리하는 단계를 포함할 수 있다.The method of treating organic waste according to another embodiment of the present invention may include treating the microbial agent to the organic waste.
본 발명의 퇴비화하는 방법에서, 상기 지오바실러스속 JH-8(Geobacillus sp. JH-8) 균주(수탁번호 KCTC13853BP)에 관한 내용은 상기 균주 및 미생물 제제에서 기재한 바와 동일하여, 명세서의 과도한 복잡성을 피하기 위하여 생략한다.In the composting method of the present invention, the content regarding the Geobacillus sp. JH-8 strain (Accession No. KCTC13853BP) is the same as described in the strain and microbial preparation, and excessive complexity of the specification is reduced omitted to avoid
본 발명의 신규한 지오바실러스속 JH-8(Geobacillus sp. JH-8) 균주 (수탁번호: KCTC13853BP)는 단백질, 탄수화물 및 지방 모두를 분해할 수 있으면서 동시에 염분에 대한 내성이 존재하기 때문에 염분이 다량으로 포함되어 있는 음식물 쓰레기 및 분뇨에서도 매우 효과적으로 성장할 수 있고, 나아가 유기물을 분해할 때 악취를 발생시키는 효소가 존재하지 않아 유기성 폐기물 분해를 위해 매우 적합하게 사용될 수 있다. 나아가, 이와 같은 균주는 배양 과정 등이 비교적 간단하여 비용을 절감시킬 수 있고, 기존에 사용되는 화학적 분해 방법에 비해 친환경 적이라는 장점이 존재한다.The novel Geobacillus genus JH-8 (Geobacillus sp. JH-8) strain of the present invention (accession number: KCTC13853BP) is capable of decomposing all proteins, carbohydrates and fats while at the same time has resistance to salt, so it contains a large amount of salt. It can grow very effectively even in food waste and manure that contain Furthermore, such a strain can reduce costs because the culture process is relatively simple, and there is an advantage of being environmentally friendly compared to the conventional chemical decomposition method.
도 1은 본 발명의 일 실시예에 따른 신규한 균주의 최적 생육 pH 범위를 그래프로 나타낸 것이다.1 is a graph showing the optimum growth pH range of a novel strain according to an embodiment of the present invention.
도 2는 본 발명의 일 실시예에 따른 신규한 균주의 최적 생육 온도 범위를 그래프로 나타낸 것이다.Figure 2 is a graph showing the optimal growth temperature range of the novel strain according to an embodiment of the present invention.
도 3은 본 발명의 일 실시예에 따른 신규한 균주의 16s rRNA를 이용한 유전형 분석 결과를 나타낸 것이다.3 shows the results of genotyping using 16s rRNA of a novel strain according to an embodiment of the present invention.
본 발명은 지오바실러스속 JH-8(Geobacillus sp. JH-8) 균주(수탁번호 KCTC13853BP), 부형제 및 식물 혼합제제를 포함하는 유기성 폐기물 분해용 미생물 제제에 관한 것이다.The present invention relates to a microbial preparation for decomposing organic waste, including a Geobacillus genus JH-8 (Geobacillus sp. JH-8) strain (Accession No. KCTC13853BP), an excipient and a plant mixture.
이하, 본 발명이 속하는 기술 분야에서 통상의 지식을 가진 자가 용이하게 실시할 수 있도록 본 발명의 실시예에 대하여 상세히 설명한다. 그러나 본 발명은 여러 가지 상이한 형태로 구현될 수 있으며 여기에서 설명하는 실시예에 한정되지 않는다.Hereinafter, embodiments of the present invention will be described in detail so that those of ordinary skill in the art can easily carry out the present invention. However, the present invention may be embodied in many different forms and is not limited to the embodiments described herein.
실시예 1Example 1
[실시예 1-1] 유기물 분해능이 우수한 균주의 선별[Example 1-1] Selection of strains with excellent organic matter degradability
50 ℃ 이상에서도 활성을 잃지 않는 균주를 아래와 같은 과정을 통해 선별하였다. 구체적으로, 경상남도 일대의 일반토양, 부엽토 및 퇴비 등에서 채취한 각종 균원 시료 1 g을 10 ml의 0.85 % NaCl 용액에 현탁하였다. 그런 다음, 상기 현탁액을 10배, 100배 및 1000배로 희석하여 영양배지(소 추출물(Beef extract) 3%, 효모 추출물(yeast extract) 5%, 아가(Agar) 1.8%, pH 7.0)가 포함된 플레이트에 도말하고, 50 ℃에서 24시간동안 배양하였다. 그런 다음, 상기 플레이트에 생성된 콜로니의 형태를 관찰한 뒤, 육안으로 보았을 때 형태가 서로 다른 콜로니를 구분하여 최종적으로 5종의 균주(이하, 'A-3', 'B-2', 'B-9', 'C-3', 'C-11'라고 함)를 선별하였다.Strains that do not lose their activity even at 50° C. or higher were selected through the following procedure. Specifically, 1 g of various fungal sources samples collected from common soil, humus, and compost around Gyeongsangnam-do were suspended in 10 ml of 0.85% NaCl solution. Then, the suspension was diluted 10-fold, 100-fold, and 1000-fold to contain a nutrient medium (beef extract 3%, yeast extract 5%, agar 1.8%, pH 7.0). Plates were plated and incubated at 50 °C for 24 hours. Then, after observing the shape of the colonies generated on the plate, colonies having different shapes when viewed with the naked eye are distinguished and finally 5 strains (hereinafter, 'A-3', 'B-2', ' B-9', 'C-3', and 'C-11') were selected.
[실시예 1-2] 선별된 균주의 성장 및 기질 용이도 확인[Example 1-2] Confirmation of growth and substrate ease of the selected strain
상기 실시예 1에서 선별된 균주의 기질에 따른 적용 가능성을 확인하기 위하여, 균주의 성장 및 효소(아밀라제(amylase), 프로테아제(protease) 및 리파아제(lipase))의 활성도를 측정하였다.In order to confirm the applicability according to the substrate of the strain selected in Example 1, the growth of the strain and the activity of enzymes (amylase, protease, and lipase) were measured.
상기 균주의 성장을 확인하기 위해, 상기 실시예 1의 균주를 영양액(nutrient broth)이 포함되어 있는 플레이트에 도말한 뒤, 50 ℃에서 12시간 배양하고, 콜로니 생성 정도에 따른 성장 속도를 확인하여 그 결과를 하기 표 1에 나타내었다. 또한, 영양 평판배지가 포함되어 있는 플레이트에 상기 실시예 1에서 선별된 균주를 접종하고 50 ℃에서 밤새도록 배양하였다. 그런 다음, 상기 배양된 균주가 존재하는 플레이트에 아이오딘(iodine)을 처리하고 투명환(clear zone)의 생성여부를 확인하여, 그 결과를 하기 표 1에 나타내었다. 여기서, 상기 영양 평판배지에는 아밀라아제의 경우 1 % 가용성 녹말(soluble starch), 프로테아제의 경우 1 %스킴 밀크(skim milk), 리파아제의 경우 1 % 트리부티린(tributyrin)가 첨가되도록 하였다.In order to confirm the growth of the strain, the strain of Example 1 was smeared on a plate containing a nutrient broth, incubated at 50 ° C. for 12 hours, and the growth rate according to the degree of colony formation was confirmed. The results are shown in Table 1 below. In addition, the strain selected in Example 1 was inoculated on a plate containing a nutrient plate medium and cultured overnight at 50 °C. Then, iodine was treated on the plate in which the cultured strain is present, and the generation of a clear zone was checked, and the results are shown in Table 1 below. Here, 1% soluble starch in the case of amylase, 1% skim milk in the case of protease, and 1% tributyrin in the case of lipase were added to the nutritional plate medium.
균주strain 성장 속도growth rate 효소 활성도Enzyme activity
아밀라제amylase 프로테아제protease 리파아제lipase
A-3A-3 ++++++ ++++++ ++++ ++++
B-2B-2 ++++++ -- -- --
B-9B-9 ++++++ ++ ++++ --
C-3C-3 ++++++ ++++ -- --
C-11C-11 ++++++ ++++ -- ++
+++ 매우 잘 자람++ 잘 자람+ 잘 자라지 않음- 없음+++ Grows very well++ Grows well + Does not grow well- None
상기 [표 1]에서 보는 바와 같이, 상기 실시예 1에서 선별된 5종의 균주는 모두 50 ℃에서 매우 잘 자랐다. 그러나, 효소 활성도의 경우에는 5종의 균주 중에서 A-3만이 아밀라제, 프로테아제 및 리파아제 효소의 활성도를 모두 가지고 있는 것을 확인하였다. 상기 결과를 통해, 본 발명에 따른 신규한 균주인 A-3의 경우에는 고온인 50 ℃에서 성장률이 매우 좋을 뿐만 아니라, 탄수화물, 단백질 및 지방을 분해할 수 있는 효소의 활성을 모두 가지고 있음을 알 수 있고, 이에 따라 상기 A-3은 유기성 폐기물을 분해에 적용할 경우 그 효율이 매우 높을 수 있음을 알 수 있다.As shown in [Table 1], all of the five strains selected in Example 1 grew very well at 50 °C. However, in the case of enzyme activity, it was confirmed that only A-3 among the five strains had all of the activities of amylase, protease and lipase enzymes. From the above results, it can be seen that in the case of A-3, a novel strain according to the present invention, not only the growth rate is very good at a high temperature of 50 °C, but also has all of the enzyme activities capable of decomposing carbohydrates, proteins and fats. and, accordingly, it can be seen that the efficiency of A-3 can be very high when the organic waste is applied to decomposition.
[실시예 1-3] 선별된 균주의 동정[Example 1-3] Identification of the selected strain
[1-3-1] 균주의 형태, 생리 및 생화학적 특성 확인[1-3-1] Confirmation of strain morphology, physiology and biochemical characteristics
상기 실시예 2에서 3종의 효소 활성을 모두 가지고 있는 A-3의 형태 등에 대해 API 키트(Biomerieux, 미국) 및 형태 분석을 수행하여, 그 결과를 하기 표 2에 나타내었다. 또한, A-3가 성장할 수 있는 최적의 pH를 확인하기 위해 상기 실시예 1의 배지에서 pH는 5.0 내지 9.0의 범위로, 온도는 30 내지 80 ℃의 범위로 조정하고, A-3를 배양한 뒤에 균체량을 측정하여, 그 결과를 도 2 및 3에 나타내었다.In Example 2, API kit (Biomerieux, USA) and morphology analysis were performed on the form of A-3 having all three enzyme activities, and the results are shown in Table 2 below. In addition, in order to confirm the optimal pH at which A-3 can grow, the pH in the medium of Example 1 was adjusted to a range of 5.0 to 9.0, and the temperature was adjusted to a range of 30 to 80 ° C., and A-3 was cultured. Subsequently, the cell mass was measured, and the results are shown in FIGS. 2 and 3 .
A-3의 식염에 대한 내성을 확인하기 위하여 상기 실시예 1과 동일한 방법으로 배양하되, 7 %의 NaCl을 포함되도록 하여 A-3의 증식 여부를 확인하였다.In order to confirm the resistance of A-3 to salt, it was cultured in the same manner as in Example 1, but it was confirmed whether or not A-3 was proliferated by including 7% NaCl.
특징Characteristic 결과 result
최적 온도(Optimum temperature)Optimum temperature 70 ℃70 ℃
최적 pH(Optimum pH)Optimum pH pH 6.5pH 6.5
색상(Pigmentation)Color (Pigmentation) 흰색White
성장(Growth) at/inGrowth at/in
β-갈락토시다제(Beta-galactosidase)β-galactosidase (Beta-galactosidase) ++
아르기닌 디하이드롤라제(Arginine dihydrolase)Arginine dihydrolase --
리신 디카르복시라제(Lysine decarboxylase)Lysine decarboxylase --
오르니틴 디카르복시라제(Ornithine decarboxylase)Ornithine decarboxylase --
시트레이트 사용 여부(Citrate utilization)Citrate utilization --
H 2S 생성H 2 S production --
우레아제(Urease)Urease --
트립토판 디아미나제(Tryptophan deaminase)Tryptophan deaminase --
인돌 생성(Indole production)Indole production --
아세토인 샌성(Acetoin production)Acetoin production ++
젤라티나제(gelatinase)Gelatinase ++
글루코스(Glucose)Glucose ++
만니톨(Mannitol)Mannitol ++
인시톨(Inositol)Inositol ++
소르비톨(Sorbitol)Sorbitol ++
람노스(Rhamnose)Rhamnose ++
수크로스(Sucrose)Sucrose ++
메리비오스(Melibiose)Melibiose ++
아미그달린(Amygdalin)Amygdalin ++
아라비노스(Arabinose)Arabinose ++
- 음성+ 양성- negative + positive
상기 [표 2]에서 보는 바와 같이, A-3는 호기성으로서, 고체배지 상에서 콜로니 형태가 원형에 가깝고 색깔은 하얀색에 해당하였으며 표면은 접적하고 매끄러운 상태임을 확인하였다. 또한, 상기 A-3는 그람 양성 간균이고, 편모가 없으며, 내생 포자를 형성하였으며, H2S, 우레아제(Urease) 및 트립토판 디아미나제(tryptophane deaminase) 등을 생산하지 않았고, 글루코스(glucose), 만니톨(mannitol) 및 수크로스(sucrose) 등과 같은 다양한 당을 이용할 수 있었다. 또한, 도 2 및 3에서 보는 바와 같이, A-3의 생육 가능 pH는 5.0 내지 9.0이었으며, 온도는 30 내지 75 ℃이었다.As shown in [Table 2], A-3 was aerobic, and it was confirmed that the colony shape on the solid medium was round, the color was white, and the surface was in contact and smooth. In addition, the A-3 is a gram-positive bacillus, has no flagella, formed endospores, did not produce H2S, urease, tryptophane deaminase, etc., and glucose, mannitol ( Various sugars such as mannitol) and sucrose were available. In addition, as shown in FIGS. 2 and 3, the viable pH of A-3 was 5.0 to 9.0, and the temperature was 30 to 75 °C.
분석 결과, A-3는 매우 높은 수준에 해당하는 7 %의 식염이 포함되어 있는 배지에서도 A-3는 매우 효과적으로 증식하였다.As a result of the analysis, A-3 proliferated very effectively even in a medium containing 7% saline, which is a very high level of A-3.
상기 결과를 통해 본 발명에 따른 A-3는 유기물을 분해할 때, 악취를 유발할 수 있는 효소를 가지고 있지 않으면서, 다양한 영양원에 의해서도 성장을 유지할 수 있을 뿐만 아니라, 유기성 폐기물 중에서 특히 음식물 쓰레기와 같이 매우 높은 수준의 식염을 포함하는 경우에도 효과적으로 증식될 수 있기 때문에, 유기성 폐기물 처리 효율을 현저하게 높일 수 있음을 알 수 있다.According to the above results, A-3 according to the present invention does not have an enzyme that can cause odor when decomposing organic matter, and can maintain growth by various nutrient sources, as well as organic waste, especially food waste. It can be seen that the organic waste treatment efficiency can be remarkably increased because it can be effectively propagated even when it contains a very high level of salt.
[1-3-2] 균주의 유전형 확인[1-3-2] Genotype confirmation of strain
GENExTM 게노믹 키트(General Biosystem, 한국)를 사용하여, 상기 A-3 균주로부터 염색체를 추출하였다. 상기 추출된 염색체와 Taq DNA 폴리머라제(Takara shuzo, 일본) 및 서열번호 2 및 3의 프라이머를 혼합하고 유전자중합효소 연쇄반응(Polymerase Chain Reaction)을 수행하여 A-3 균주의 16s rRNA를 증폭한 뒤, ABI PRISM 310 genetic analyzer(Perkin-Elmer Applied Biosystems, 미국)을 사용하여 염기서열을 분석하였다. 또한, 상동성 분석의 경우에는 NCBI BLAST 프로그램을 이용하여, 그 결과를 도 1에 나타내었다.Chromosomes were extracted from the A-3 strain using the GENEx™ genomic kit (General Biosystem, Korea). After mixing the extracted chromosome with Taq DNA polymerase (Takara shuzo, Japan) and the primers of SEQ ID NOs: 2 and 3, and performing Polymerase Chain Reaction to amplify 16s rRNA of strain A-3, , The nucleotide sequence was analyzed using an ABI PRISM 310 genetic analyzer (Perkin-Elmer Applied Biosystems, USA). In addition, in the case of homology analysis, the NCBI BLAST program was used, and the results are shown in FIG. 1 .
하기의 도 1에서 보는 바와 같이, A-3 균주는 서열번호 1의 16s rRNA 서열을 가지며, 지오바실러스 잘리헤(Geobacillus zalihae) NBRC 101842와 가장 높은 99 %의 서열 상동성을 나타냄을 확인하였다.As shown in FIG. 1 below, it was confirmed that strain A-3 has the 16s rRNA sequence of SEQ ID NO: 1, and exhibits the highest 99% sequence homology with Geobacillus zalihae NBRC 101842.
상기 [1-3-1] 및 [1-3-2]의 결과를 종합한 결과, A-3는 지오바실러스 속에 속하는 신규한 지오바실러스 속 JH-8(Geobacillus sp. JH-8) 균주로 동정하고, 이를 한국생명공학연구원에 기탁하였다(기탁번호: KCTC13853BP).As a result of synthesizing the results of [1-3-1] and [1-3-2], A-3 was identified as a novel Geobacillus genus JH-8 (Geobacillus sp. JH-8) strain belonging to the genus Geobacillus. and deposited it with the Korea Research Institute of Bioscience and Biotechnology (Accession No.: KCTC13853BP).
실험예 2Experimental Example 2
[실험예 2-1] 미생물 제제의 유기성 폐기물에 대한 분해 활성 평가 1[Experimental Example 2-1] Evaluation of decomposition activity on organic waste of microbial preparations 1
상기 지오바실러스속 JH-8(Geobacillus sp. JH-8) 균주를 이용하여 유기성 폐기물을 분해하기 위한 미생물 제제(JH8)를 제조하였다. 그에 따른 활성을 평가하기 위하여 하기의 축산분뇨(A), 음식폐기물(B)에 상기 미생물 제제를 처리하고 악취가 제거되는데 소용되는 시간(분해율 1, E1) 및 중량이 절반으로 감소하는데 걸리는 시간(분해율 2, E2)을 평가하였다.A microbial preparation (JH8) for decomposing organic waste was prepared using the Geobacillus genus JH-8 (Geobacillus sp. JH-8) strain. In order to evaluate the resulting activity, the time it takes for the microbial preparation to be treated with the following livestock manure (A) and food waste (B) and the odor is removed (decomposition rate 1, E1) and the time it takes for the weight to be reduced by half ( The degradation rate 2, E2) was evaluated.
대조군으로는 상기 축산분뇨(A)에 미생물 제제를 처리하지 않는 경우(Con)로 하여 상기 대조군과 비교하여 상기 미생물 제제가 처리된 경우를 비교하여 그 평가결과를 지수로 평가하였다. 해당 지수는 그 숫자가 낮을수록 분해 효율이 우수한 것이다.As a control, the case where the microbial preparation was not treated (Con) in the livestock manure (A) was compared with the control and the microbial preparation was treated, and the evaluation result was evaluated as an index. In this index, the lower the number, the better the decomposition efficiency.
이를 하기의 [표 3]에 나타내었다.This is shown in [Table 3] below.
Con(A)Con(A) A+JH8A+JH8 B+JH8B+JH8
E 1E 1 1010 22 33
E 2E 2 1010 44 55
상기 [표 3]을 참조하면, 본 발명에 따른 미생물 제제를 사용하는 경우 분뇨 또는 음식물 폐기물과 같은 유기성 폐기물을 분해하는 활성이 우수하다는 사실을 확인할 수 있다. 특히 염분이 포함된 음식물 폐기물에 대하여 악취 제거 및 분해활성이 우수하다는 점을 알 수 있어, 본 발명에 제시하는 미생물 제제의 미생물은 염분에 대한 저항성이 높다는 점을 알 수 있다.Referring to [Table 3], it can be confirmed that when the microbial preparation according to the present invention is used, the activity of decomposing organic waste such as manure or food waste is excellent. In particular, it can be seen that the odor removal and decomposition activity for food waste containing salt is excellent, and it can be seen that the microorganism of the microbial preparation presented in the present invention has high resistance to salt.
[실험예 2-2] 미생물 제제의 유기성 폐기물에 대한 분해 활성 평가 2[Experimental Example 2-2] Evaluation of decomposition activity on organic waste of microbial preparations 2
상기 미생물 제제에 대하여 식물 혼합제제를 혼합함으로서 유기성 폐기물에 대한 분해 활성이 증진 정도를 평가하기 위하여 하기의 [표 4]와 같은 식물 추출물을 제조하였다.Plant extracts as shown in [Table 4] below were prepared in order to evaluate the degree of enhancement of decomposition activity for organic wastes by mixing plant mixtures with respect to the microbial preparations.
한편, 오미자 추출물(S), 창포 추출물(T), 가지 잎 추출물(U), 민들레 추출물(X), 노간주나무 추출물(Z)는 각 식물 소재를 열수추출 및 여과하여 제조하였다. 또한 상기 식물 추출물을 하기의 [표 4]와 같이 혼합하고 상기 미생물 제제에 혼합하여 유기성 폐기물 분해용 미생물 제제를 제조하였다.On the other hand, Schisandra extract (S), iris extract (T), eggplant leaf extract (U), dandelion extract (X), and juniper extract (Z) were prepared by hot water extraction and filtration of each plant material. In addition, the plant extract was mixed as shown in Table 4 below and mixed with the microbial preparation to prepare a microbial preparation for decomposing organic waste.
M1M1 M2M2 M3M3 M4M4 M5M5 M6M6 M7M7 M8 M8 M9M9
SS 100100 100100 100100 100100 100100 100100
T T 100100 0.10.1 1One 55 1010 1515
U U 100100 0.10.1 1One 55 1010 1515
X X 100100 0.10.1 0.50.5 33 55 77
ZZ 0.010.01 0.10.1 1One 22 33
(단위: 중량부)상기 제조된 미생물 제제를 분뇨(A)에 처리하여 상술한 방법과 마찬가지고 분해율 1(E1) 및 분해율 2(E2)를 평가하였다. 식물 혼합제제에 따른 상승효과를 확인하기 위하여 식물 혼합제제를 사용하지 않고 미생물 제제만 사용한 경우를 대조군으로 하여 10으로 그 지수를 고정하고, 상기 식물 혼합제제가 혼합된 실시예를 비교 평가하여 지수로 그 결과를 나타내었다. 상기 지수는 그 숫자가 낮을수록 효과가 우수한 것이다.(Unit: parts by weight) The prepared microbial preparation was treated in the manure (A), and the degradation rate 1 (E1) and degradation rate 2 (E2) were evaluated in the same manner as in the above-described method. In order to confirm the synergistic effect according to the plant mixture preparation, the index was fixed to 10 as a control when only a microbial preparation was used without using a plant mixture preparation, Comparative evaluation of examples in which the plant mixture preparation was mixed was evaluated as an index The results are shown. In the index, the lower the number, the better the effect.
그 결과를 하기의 [표 5]에 나타내었다.The results are shown in [Table 5] below.
JH8JH8 JH8+M1JH8+M1 JH8+M2JH8+M2 JH8+M3JH8+M3 JH8+M4JH8+M4 JH8+M5JH8+M5 JH8+M6JH8+M6 JH8+M7JH8+M7 JH8+M8JH8+M8 JH8+M9JH8+M9
E1E1 1010 88 1010 1010 99 99 77 66 66 99
E2E2 1010 99 1010 1010 1010 1010 88 77 77 1010
(단위: 지수)(Unit: Index)
상기 [표 5]를 참조하는 경우, 오미자 추출물을 단독으로 사용하는 경우 악취를 제거하는 효과가 빠르게 나타났다. 또한 오미자 추출물과 다른 식물 추출물을 혼합하는 경우 M5 내지 M8의 범위에 의하는 경우 각 식물 추출물의 혼합에 따른 상승효과로서 유기성 폐기물의 분해효과 및 악취제거효과가 높아진다는 사실을 확인할 수 있다.Referring to [Table 5], when the Schisandra extract was used alone, the effect of removing the odor was quickly shown. In addition, when the Schisandra extract and other plant extracts are mixed, according to the range of M5 to M8, it can be confirmed that the decomposition effect of organic waste and the odor removal effect are increased as a synergistic effect according to the mixing of each plant extract.
따라서 상기 미생물에 제제에 대하여 일정한 범위의 식물 추출물을 혼합하여 식물 혼합 제제로 혼합하여 사용하는 경우 미생물의 분해활성을 높일 수 있다는 점을 알 수 있다.Therefore, it can be seen that the decomposition activity of microorganisms can be increased when the microorganisms are mixed with a plant extract of a certain range with respect to the preparation and used as a plant mixture preparation.
이상으로 본 발명의 특정한 부분을 상세히 기술하였는 바, 당업계의 통상의 지식을 가진 자에게 있어서 이러한 구체적인 기술은 단지 바람직한 구현 예일 뿐이며, 이에 본 발명의 범위가 제한되는 것이 아닌 점은 명백하다. 따라서, 본 발명의 실질적인 범위는 첨부된 청구항과 그의 등가물에 의하여 정의된다고 할 것이다.As described above in detail a specific part of the present invention, for those of ordinary skill in the art, this specific description is only a preferred embodiment, and it is clear that the scope of the present invention is not limited thereto. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.
[수탁번호][Accession Number]
기탁기관명 : 한국생명공학연구원Name of deposit institution: Korea Research Institute of Bioscience and Biotechnology
수탁번호 : KCTC13853BPAccession number: KCTC13853BP
수탁일자 : 20190603Deposit date: 20190603
본 발명은 신규한 지오바실러스속 균주 및 부형제를 포함하는 유기성 폐기물 분해용 미생물 제제에 관한 것이다. 보다 상세하게는 토양 등에서 분리한 신규한 지오바실러스속 균주의 유기성 폐기물의 분해 활성을 이용하여 유기성 폐기물을 분해하거나, 유기성 폐기물을 퇴비화할 수 있는 미생물 제제를 제공하고, 유기성 폐기물을 분해하거나 퇴비화시키는 방법을 제공하기 위한 것이다.The present invention relates to a microorganism preparation for decomposing organic waste comprising a novel strain of the genus Geobacillus and an excipient. More specifically, a method of decomposing organic waste using the decomposition activity of organic wastes of a novel Geobacillus sp. strain isolated from soil, etc., or providing a microbial agent capable of composting organic waste, decomposing or composting organic waste is to provide
서열목록 전자파일 첨부(C:\Users\jcjung\Desktop\WP193073_동의대_신규한 지오바실러스속 균주 및 이의 용도\지오바실러스-서열목록.app)Attached sequence list electronic file (C:\Users\jcjung\Desktop\WP193073_Donguidae_new strains of the genus Geobacillus and its use\Giobacillus-sequence list.app)

Claims (9)

  1. 지오바실러스속 JH-8(Geobacillus sp. JH-8) 균주(수탁번호 KCTC13853BP), 부형제 및 식물 혼합제제를 포함하는Geobacillus genus JH-8 (Geobacillus sp. JH-8) strain (accession number KCTC13853BP), including excipients and plant mixtures
    유기성 폐기물 분해용 미생물 제제.Microbial preparations for the decomposition of organic waste.
  2. 제 1항에 있어서,The method of claim 1,
    상기 균주는 아밀라제(amylase), 프로테아제(protease) 및 리파아제(lipasThe strain is amylase (amylase), protease (protease) and lipase (lipas)
    e)으로 구성된 군으로부터 선택되는 1종 이상의 효소 활성이 있는 것인e) having at least one enzymatic activity selected from the group consisting of
    유기성 폐기물 분해용 미생물 제제.Microbial preparations for the decomposition of organic waste.
  3. 제 1항에 있어서,The method of claim 1,
    상기 균주는 염분 내성이 있는 것인The strain is salt tolerant
    유기성 폐기물 분해용 미생물 제제.Microbial preparations for the decomposition of organic waste.
  4. 제 1항에 있어서,The method of claim 1,
    상기 균주는 최적 생장 온도가 30 내지 80 ℃이고, 최적 생장 pH가 5 내지 9 인 것인The strain has an optimal growth temperature of 30 to 80 °C, and an optimal growth pH of 5 to 9
    유기성 폐기물 분해용 미생물 제제.Microbial preparations for the decomposition of organic waste.
  5. 제 1항에 있어서,The method of claim 1,
    상기 유기성 폐기물은 음식물 쓰레기 또는 분뇨인 것인The organic waste will be food waste or manure
    유기성 폐기물 분해용 미생물 제제.Microbial preparations for the decomposition of organic waste.
  6. 지오바실러스속 JH-8(Geobacillus sp. JH-8) 균주(수탁번호 KCTC13853BP), 부형제 및 식물 혼합제제를 포함하는Geobacillus genus JH-8 (Geobacillus sp. JH-8) strain (accession number KCTC13853BP), including excipients and plant mixtures
    유기성 폐기물의 퇴비화를 위한 미생물 제제.Microbial preparations for composting of organic waste.
  7. 제 6항에 있어서,7. The method of claim 6,
    상기 유기성 폐기물은 음식물 쓰레기 또는 분뇨인 것인The organic waste will be food waste or manure
    유기성 폐기물의 퇴비화를 위한 미생물 제제.Microbial preparations for composting of organic waste.
  8. 제 6항 또는 제 7항 중 어느 한 항의 미생물 제제를 유기성 폐기물에 처리하는 단계를 포함하는8. A method comprising the step of treating the microbial agent of any one of claims 6 or 7 to organic waste.
    유기성 폐기물을 퇴비화하는 방법.How to compost organic waste.
  9. 제 1항 내지 제 5항 중 어느 한 항의 미생물 제제를 유기성 폐기물에 처리하는 단계를 포함하는Claims 1 to 5 comprising the step of treating the microbial agent of any one of claims to organic waste
    유기성 폐기물을 처리하는 방법.How to dispose of organic waste.
PCT/KR2020/006370 2019-12-06 2020-05-14 Microbial preparation for decomposing organic wastes comprising novel geobacillus sp. strain and excipient WO2021112349A1 (en)

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Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20050086014A (en) * 2003-08-27 2005-08-30 가부시키가이샤 신꼬간꾜우솔루션 New microorganism and method of treating organic solid materials by using the same
JP2006238794A (en) * 2005-03-03 2006-09-14 Tokyo Gas Co Ltd New microorganism and method for treating wastewater and apparatus for treating wastewater using the new microorganism
KR100807275B1 (en) * 2006-12-22 2008-02-28 목원대학교 산학협력단 Hyperthermophilic protease-producing geobacillus kaustophilus c-2 strain and method using the same
KR20090126917A (en) * 2008-06-05 2009-12-09 한경대학교 산학협력단 Novel geobacillus thermodenitrificance sg-01 having nitrates-eliminating function and method using the same
KR20110002512A (en) * 2009-07-02 2011-01-10 학교법인 동의학원 Geobacillus sp. hw1 strain producing thermophilic lipase and usage thereof
KR102055544B1 (en) * 2019-06-11 2019-12-13 동의대학교 산학협력단 A novel Geobacillus sp. strain and use thereof

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20050086014A (en) * 2003-08-27 2005-08-30 가부시키가이샤 신꼬간꾜우솔루션 New microorganism and method of treating organic solid materials by using the same
JP2006238794A (en) * 2005-03-03 2006-09-14 Tokyo Gas Co Ltd New microorganism and method for treating wastewater and apparatus for treating wastewater using the new microorganism
KR100807275B1 (en) * 2006-12-22 2008-02-28 목원대학교 산학협력단 Hyperthermophilic protease-producing geobacillus kaustophilus c-2 strain and method using the same
KR20090126917A (en) * 2008-06-05 2009-12-09 한경대학교 산학협력단 Novel geobacillus thermodenitrificance sg-01 having nitrates-eliminating function and method using the same
KR20110002512A (en) * 2009-07-02 2011-01-10 학교법인 동의학원 Geobacillus sp. hw1 strain producing thermophilic lipase and usage thereof
KR102055544B1 (en) * 2019-06-11 2019-12-13 동의대학교 산학협력단 A novel Geobacillus sp. strain and use thereof

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