WO2024058613A1 - Novel azospirillum sp. fwr9 and compost using same - Google Patents
Novel azospirillum sp. fwr9 and compost using same Download PDFInfo
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- WO2024058613A1 WO2024058613A1 PCT/KR2023/013953 KR2023013953W WO2024058613A1 WO 2024058613 A1 WO2024058613 A1 WO 2024058613A1 KR 2023013953 W KR2023013953 W KR 2023013953W WO 2024058613 A1 WO2024058613 A1 WO 2024058613A1
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- azospirillum
- compost
- fwr9
- kacc
- food
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Classifications
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09B—DISPOSAL OF SOLID WASTE
- B09B3/00—Destroying solid waste or transforming solid waste into something useful or harmless
- B09B3/60—Biochemical treatment, e.g. by using enzymes
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
- C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
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- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
-
- 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
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/30—Fuel from waste, e.g. synthetic alcohol or diesel
-
- 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 provides a novel Azospirillum sp. suitable for use in compost production.
- Azospirillum sp. FWR9 (KACC 81197BP) and compost using it.
- Coffee has been used as a favorite food in various countries. In Korea, the annual consumption of coffee per person is 338 cups, which is a large amount, and coffee consumption is increasing year by year. Meanwhile, coffee has a problem in that more than 90% of it is discarded in the form of grounds (coffee grounds) during the manufacturing process. In addition, coffee grounds are discarded as general household waste or food waste, which causes methane gas in the process, and in particular, those discarded as domestic wastewater flow into sewers and pollute water quality. Additionally, it has been reported that landfilled coffee grounds emit methane gas, which is more than 20 times more likely to cause global warming than carbon dioxide. Nevertheless, it is known that only 0.1% of coffee grounds are reused.
- Methods for recycling food waste include feed conversion, composting, and biogasification through anaerobic digestion.
- Various research institutes and companies are researching the treatment of food waste, but currently operating feed and compost recycling facilities are causing problems such as bad odor, wastewater discharge, excessive energy use, and greenhouse gas emissions.
- a method is needed to reduce food-related waste such as coffee grounds and food waste.
- a novel Azospirillum sp. can be used to produce compost useful for crop cultivation from food-related waste.
- the present invention relates to Azospirillum sp. having trypsin activity.
- Azospirillum sp. FWR9 (KACC 81197BP) is provided.
- the present invention relates to Azospirillum sp. having trypsin activity.
- a microbial preparation for composting of food-related waste comprising ( Azospirillum sp.) FWR9 (KACC 81197BP).
- the present invention relates to Azospirillum sp. having trypsin activity.
- Azospirillum sp. Provides compost that is manufactured by mixing FWR9 (KACC 81197BP) and food-related waste.
- the compost of the present invention is preferably Neorizobium sp. ( Neorhizobium sp.) FWR7 (KACC 81196BP) and Microbacterium testaceum ( Microbacterium testaceum ) FWT1 (KACC 81195BP) are preferably prepared by further mixing one or more microorganisms selected from among them.
- Neorizobium sp. Neorhizobium sp.
- FWR7 KACC 81196BP
- Microbacterium testaceum Microbacterium testaceum
- FWT1 KACC 81195BP
- the food-related waste preferably contains food waste in a weight ratio of 4 to 6, coffee grounds in a weight ratio of 4 to 6, and rice husk in a weight ratio of 0.8 to 1.2.
- Azospirillum sp of the present invention Azospirillum sp.
- FWR9 KACC 81197BP
- Compost using FWR9 (KACC 81197BP) allows crops to grow well.
- Figure 1 shows that in order to isolate strains of the present invention, samples were obtained from a food composting device and then cultured on a plate.
- Figure 2 shows the present invention Neorizobium sp. ( Neorhizobium sp.) This is the result of phylogenetic analysis of FWR7 (KACC 81196BP).
- FIG 3 shows the present invention Azospirillum sp. ( Azospirillum sp.) This is the result of phylogenetic analysis of FWR9 (KACC 81197BP).
- Figure 4 is a phylogenetic analysis result of Microbacterium testaceum FWT1 (KACC 81195BP) of the present invention.
- Figure 5 shows the present invention Neorizobium sp. ( Neorhizobium sp.) FWR7 (KACC 81196BP), Azospirillum sp. ( Azospirillum sp.) FWR9, Microbacterium testaceum FWT1 microscopic observation results.
- Figure 6 shows the present invention Neorizobium sp. ( Neorhizobium sp.) This is the result of analysis of enzyme activity and physiological characteristics of FWR7 (KACC 81196BP).
- Figure 7 shows the present invention Azospirillum sp. ( Azospirillum sp.) This is the result of analysis of enzyme activity and physiological characteristics of FWR9 (KACC 81197BP).
- Figure 8 shows the results of analysis of enzyme activity and physiological characteristics of Microbacterium testaceum FWT1 (KACC 81195BP) of the present invention.
- Figure 9 shows the primary fermentation compost obtained by mixing coffee waste, food waste, and rice husk and fermenting for 2 days, mixing it with general topsoil in a weight ratio of 1:1, then cultivating lettuce for 30 days, and primary fermentation compost. This is the result of comparing the crop growth promotion ability of with general soil.
- Figure 10 shows the primary fermentation compost, Neorizobium sp. of the present invention.
- Neorhizobium sp. Freeze-dried powder (mixed strains) containing FWR7 (KACC 81196BP) was added and fermented for 30 days to prepare secondary fermentation compost, and the secondary fermentation compost was mixed with general topsoil 1:1, 2 :1 This is the result of growing lettuce for 30 days after mixing at a weight ratio, and comparing the crop growth promotion ability of secondary fermentation compost with general topsoil.
- the present invention relates to Azospirillum sp. ( Azospirillum sp.) FWR9 (KACC 81197BP) is provided.
- Azospirillum sp of the present invention was isolated from a food waste composting device and has the 16S rRNA sequence of SEQ ID NO: 2.
- SEQ ID NO: 2 16S rRNA sequence of SEQ ID NO: 2.
- Azospirillum sp of the present invention is good for use in producing compost useful for crop cultivation.
- Azospirillum sp of the present invention is good for use in producing compost useful for crop cultivation.
- Azospirillum sp.) FWR9 (KACC 81197BP) exhibits various enzyme activities and physiological characteristics. It decomposes the high molecular substances contained in compost into low molecular substances, allowing crops to more easily absorb useful substances contained in the compost. Additionally, it is involved in the nitrogen cycle and can help crop growth.
- Azospirillum sp. of the present invention Compost fermented using FWR9 (KACC 81197BP) has excellent crop growth promotion ability.
- the present invention is Azospirillum sp.
- a microbial preparation for composting of food-related waste using ( Azospirillum sp.) FWR9 (KACC 81197BP) is provided.
- the microbial agent is Azospirillum sp. of the present invention.
- Azospirillum sp. This means that it contains FWR9 (KACC 81197BP), and as an example, the strain may be manufactured in the form of dried powder.
- the microbial preparation prepared in this way can be used by injecting it into a compost manufacturing device.
- the food-related waste refers to waste such as slaughter waste from a slaughterhouse, waste from a food processing plant, and food waste discharged from homes.
- the present invention is Azospirillum sp.
- Azospirillum sp. Provides compost that is manufactured by mixing FWR9 (KACC 81197BP) and food-related waste.
- Neorizobium sp. Neorhizobium sp.
- FWR7 KACC 81196BP
- microorganisms selected from Microbacterium testaceum FWT1 KACC 81195BP
- Azospirillum sp. Azospirillum sp.
- FWR9, FWR7, and FWT1 break down food-related waste into small pieces, helping crops and other plants more easily absorb organic substances contained in compost. Therefore, when the compost prepared as above is used for crop cultivation, the crops grow well. Meanwhile, FWR9, FWR7 and FWT1 are preferably mixed and used in a ratio of 0.8 to 1.2:0.8 to 1.2:0.8 to 1.2, respectively.
- Food waste, coffee waste, and rice husk that are discarded in landfills can be further utilized to help resource circulation.
- coffee grounds are mainly household waste generated at coffee shops, and the recycling rate is known to be very low, around 0.1%.
- the compost of the present invention may be manufactured by adding other supplementary ingredients during production.
- Preferred examples include bone meal, sake lees, and seed cake.
- strains suitable for compost production were selected. Through this, it was confirmed that strains FWR9, FWT1, and FWR7 were excellent at composting food-related waste.
- the 16S rRNA sequence of SEQ ID NO: 1 was obtained.
- the sequence was approximately 97.3% similar to Neorhizobium tomejilense .
- Figure 2 it was confirmed that it was a new strain distinct from other species within the existing genus.
- Table 1 it was confirmed that the similarity index was low compared to the previously known strains.
- the FWR7 strain of the present invention is a novel Neorizobium sp. It was found that it was a type of strain ( Neorhizobium sp.).
- the FWR9 strain For the FWR9 strain, the 16S rRNA sequence of SEQ ID NO: 2 was obtained. As a result of genetic identification using SEQ ID No. 2, it was confirmed that the sequence was approximately 98.1% similar to Azospirillum thiophilum . In addition, through phylogenetic analysis (Figure 3), it was confirmed that it was distinguished from other species within the existing genus. In addition, as a result of analyzing the fatty acid characteristics of the membrane of the FWR9 strain (Table 2), it was confirmed that the similarity index was low compared to the previously known strains. Based on this, the FWR9 strain of the present invention is a new Azospirillum sp. ( Azospirillum sp.) It was found to be a type of strain.
- the 16S rRNA sequence of SEQ ID NO: 3 was obtained. As a result of genetic identification using SEQ ID NO: 3, it was confirmed that the sequence was approximately 99.13% similar to Microbacterium testaceum . In addition, through phylogenetic analysis (FIG. 4), it was found to be a Microbacterium testaceum strain.
- Neorizobium sp. Neorhizobium sp.
- FWR7 KACC 81196BP
- Microbacterium testaceum FWT1 KACC 81195BP
- Azospirillum sp. Azospirillum sp. was named FWR9 (KACC 81197BP).
- Neorizobium sp of the present invention was 20 ⁇ 37°C, the optimal culture temperature was 25°C, the suitable culture salinity was 0 ⁇ 2.0%, and the suitable culture pH was 7 ⁇ 8.
- Azospirillum sp of the present invention As a result of cultivating FWR9, it was confirmed that the suitable culture temperature was 20 ⁇ 37°C, the optimal culture temperature was 25°C, the suitable culture salinity was 0 ⁇ 1.5%, and the suitable culture pH was 7 ⁇ 8.
- the suitable culture temperature was 20 to 37 °C
- the optimal culture temperature was 25 °C
- the suitable culture salinity was 0 to 2.0%
- the suitable culture pH was 7 to 7. I was able to confirm that it was 8.
- Neorizobium sp. Neorhizobium sp.
- Estrease C4
- Estrease Lipase CB
- Leucine arylamidase Trypsin, ⁇ -chymotrypsin, acid phospatase, Naphtol-AS-Bl-phosphohydrolase
- beta-glucuronidase ⁇ -glucurorndase
- alpha-glucosidase beta-glucosidase
- beta-glucosidase N-acetyl-beta-glucosaminindase were positive. appeared (a in Figure 6).
- Neorizobium sp. Neorhizobium sp.
- arginine dihydrolase urease, hydrolysis (b-glucosidase), beta-glucosidase, and glucose assimilation.
- assimilation, arabinose assimilation, mannose assimilation, mannitol assimilation, N-acetyl-glucosamine assimilation, maltose assimilation, gluconate Gluconate assimilation and malate assimilation were positive (Figure 6b).
- Neorhizobium sp. The results of measuring enzyme activity and physiological characteristics of FWR7 are Neorhizobium sp. of the present invention.
- Neorhizobium sp. This means that when producing compost using FWR7, high molecular substances (carbohydrates, fats, proteins) can be converted into low molecular substances to help crop growth.
- Azospirillum sp. ( Azospirillum sp.) As a result of testing the enzyme activity of FWR9, alkaline phosphatase, Estrease (C4), Estrease Lipase (CB), Leucine arylamidase, Trypsin, acid phospatase, Naphtol-AS-Bl-phosphohydrolase, beta-glucosidase, alpha-fucosidase ( ⁇ -fucosidase) was found to be positive (Figure 7a).
- Azospirillum sp. Azospirillum sp.
- a-glucosidase As a result of testing the physiological properties of FWR9, it was found that duction of nitrates, arginine dihydrolase, urease, ⁇ -glucosidase, beta- Galactosidase, glucose assimilation, mannitol assimilation, and N-acetyl-glucosamine assimilation were positive (Figure 7b).
- the enzyme activity measurement results and physiological characteristics measurement results of FWR9 are the Azospirillum sp. of the present invention.
- the enzyme activity measurement results and physiological characteristics measurement results of Microbacterium testaceum FWT1 are, when producing compost using Microbacterium testaceum FWT1 of the present invention, the polymer This means that substances (carbohydrates, fats, proteins) can be converted into low-molecular substances to help crops grow.
- the food waste used in the present invention was obtained from a food composting device. Specifically, the food composting device processes food using a dilution fermentation method. It is a device that ferments food to reduce its weight, dehydrates it, and then collects the remaining food waste in a recovery device. In the present invention, two days after putting food into the food composting device, the food scraps collected in the recovery device were used as food waste for compost production.
- Compost using food waste and coffee grounds could be produced within the recovery device. Specifically, it was manufactured by putting coffee grounds and rice husk into a recovery device containing food waste and fermenting it for 2 days. The specific addition ratio is shown in Table 3 below.
- Preparation Example 5 After preparing Preparation Example 5, the compost ingredients were tested and the moisture content was found to be appropriate, making it suitable for use as compost. In addition, the phosphoric acid and potassium components were higher than those of Preparation Example 4, making it more suitable for crop cultivation.
- Microorganisms were added to the first fermentation compost and fermented compost was prepared.
- primary fermentation compost was prepared by mixing 5 kg of coffee grounds, 5 kg of food waste, and 1 kg of rice husk by the method described in 3-2) above. After mixing 0.1 g of mixed strains with the prepared primary fermentation compost, secondary fermentation was performed for 30 days to prepare secondary fermentation compost.
- Neorizobium sp. of the present invention described in Example 1.
- Neorhizobium sp. FWR7, Azospirillum sp.
- Azospirillum sp. FWR9
- Microbacterium testaceum Microbacterium testaceum
- the prepared secondary fermentation compost and general topsoil were mixed at a ratio of 1:1 and 2:1, respectively, and then lettuce was grown for 30 days (Figure 10), and root growth, leaf size, etc. Comprehensive comparison was made.
- Azospirillum sp. of the present invention This means that the mixed strain including FWR9 (KACC 81197BP) decomposed the organic materials of food-related waste used in compost production into an appropriate size, converting them to an appropriate size for crops to absorb.
- FWR9 KACC 81197BP
Abstract
The present invention relates to novel Azospirillum sp. FWR9 (KACC 81197BP), which is good for use in compost preparation, and compost using same. Azospirillum sp. FWR9 (KACC 81197BP) of the present invention degrades high molecular weight organic materials (carbohydrates, proteins, fats and the like) into appropriate sizes so that organic materials are converted and easily absorbed by crops. Therefore, compost using Neorhizobium sp. FWR7 (KACC 81196BP) of the present invention promotes crop growth.
Description
본 발명은 퇴비 제조에 사용하기 좋은 신규의 아조스피릴룸 sp. (Azospirillum sp.) FWR9 (KACC 81197BP) 및 이를 이용한 퇴비에 관한 것이다.The present invention provides a novel Azospirillum sp. suitable for use in compost production. ( Azospirillum sp.) FWR9 (KACC 81197BP) and compost using it.
커피는 다양한 나라에서 기호 식품으로 이용되어 왔다. 우리나라에서는 1인당 1년 커피 소비량이 338잔으로, 많은 량이 소비되고 있으며, 커피의 소비는 해가 갈수록 증가 하고 있다. 한편, 커피는 제조 과정 중에 90% 이상을 찌꺼기 형태 (커피박)로 버려지는 문제점이 있다. 또한, 커피박은 일반 생활 폐기물 또는 음식 폐기물로 버려지는데, 그 과정에서 메탄가스를 유발시키고, 특히, 생활폐수로 버려진 것은 하수구로 흘러들어 수질을 오염시키고 있다. 또한, 매립된 커피박은 이산화탄소 보다 지구온난화를 일으킬 위험이 20배 이상 높은 메탄가스를 배출한다고 보고된 바 있다. 그럼에도 커피박은 오직 0.1% 만이 재사용되는 것으로 알려져 있다. Coffee has been used as a favorite food in various countries. In Korea, the annual consumption of coffee per person is 338 cups, which is a large amount, and coffee consumption is increasing year by year. Meanwhile, coffee has a problem in that more than 90% of it is discarded in the form of grounds (coffee grounds) during the manufacturing process. In addition, coffee grounds are discarded as general household waste or food waste, which causes methane gas in the process, and in particular, those discarded as domestic wastewater flow into sewers and pollute water quality. Additionally, it has been reported that landfilled coffee grounds emit methane gas, which is more than 20 times more likely to cause global warming than carbon dioxide. Nevertheless, it is known that only 0.1% of coffee grounds are reused.
한편, 환경공단의 2019년도 발표에 따르면, 생활 폐기물 중 음식물 폐기물은 하루 16,000톤 배출되어 전체 생활 폐기물의 약 30%를 차지하는데, 연간 처리 비용만 8,000억원으로 음식물 폐기물의 자원화에 대한 중요성이 부각되고 있다. Meanwhile, according to the Korea Environment Corporation's 2019 announcement, 16,000 tons of food waste is produced per day, accounting for about 30% of the total household waste, and the annual disposal cost alone is 800 billion won, highlighting the importance of turning food waste into a resource. there is.
음식물 폐기물의 자원화 처리방법에는 사료화, 퇴비화, 혐기성 소화를 통한 바이오가스화 처리방법이 있다. 다양한 연구기관 및 회사에서 음식 폐기물에 대한 처리에 연구 중에 있으나, 현재 운영되고 있는 사료 및 퇴비 자원화 처리 시설은 악취 발생, 폐수 배출, 과도한 에너지 사용, 온실가스 배출 등의 문제를 발생시키고 있다.Methods for recycling food waste include feed conversion, composting, and biogasification through anaerobic digestion. Various research institutes and companies are researching the treatment of food waste, but currently operating feed and compost recycling facilities are causing problems such as bad odor, wastewater discharge, excessive energy use, and greenhouse gas emissions.
커피박, 음식물 폐기물 등의 식품 관련 폐기물을 저감화시킬 수 있는 방법이 필요하다. A method is needed to reduce food-related waste such as coffee grounds and food waste.
이에 본 발명에서는 식품 관련 폐기물로부터 작물 재배에 유용한 퇴비를 제조할 수 있는 신규의 아조스피릴룸 sp. (Azospirillum sp.) FWR9 (KACC 81197BP)를 제공하고자 한다. Accordingly, in the present invention, a novel Azospirillum sp. can be used to produce compost useful for crop cultivation from food-related waste. We would like to provide ( Azospirillum sp.) FWR9 (KACC 81197BP).
또한, 본 발명에서는 식품 관련 폐기물 및 본 발명의 아조스피릴룸 sp. (Azospirillum sp.) FWR9 (KACC 81197BP)를 사용하여 제조한 퇴비를 제공하고자 한다.In addition, in the present invention, food-related waste and Azospirillum sp of the present invention. We would like to provide compost manufactured using ( Azospirillum sp.) FWR9 (KACC 81197BP).
본 발명은 트립신(Trypsin) 활성을 가지는 아조스피릴룸 sp. (Azospirillum sp.) FWR9 (KACC 81197BP)을 제공한다.The present invention relates to Azospirillum sp. having trypsin activity. ( Azospirillum sp.) FWR9 (KACC 81197BP) is provided.
또한, 본 발명은 트립신(Trypsin) 활성을 가지는 아조스피릴룸 sp. (Azospirillum sp.) FWR9 (KACC 81197BP)를 포함하는 것을 특징으로 하는 식품 관련 폐기물의 퇴비화를 위한 미생물 제제를 제공한다.In addition, the present invention relates to Azospirillum sp. having trypsin activity. Provided is a microbial preparation for composting of food-related waste, comprising ( Azospirillum sp.) FWR9 (KACC 81197BP).
또한, 본 발명은 트립신(Trypsin) 활성을 가지는 아조스피릴룸 sp. (Azospirillum sp.) FWR9 (KACC 81197BP) 및 식품 관련 폐기물을 혼합하여 제조한 것을 특징으로 하는 퇴비를 제공한다.In addition, the present invention relates to Azospirillum sp. having trypsin activity. ( Azospirillum sp.) Provides compost that is manufactured by mixing FWR9 (KACC 81197BP) and food-related waste.
한편, 상기 본 발명의 퇴비는 바람직하게 네오리조비움 sp. (Neorhizobium sp.) FWR7 (KACC 81196BP), 마이크로박테리움 테스타시움 (Microbacterium testaceum) FWT1 (KACC 81195BP) 중 선택되는 어느 하나 이상의 미생물을 더욱 혼합하여 제조한 것이 좋다.Meanwhile, the compost of the present invention is preferably Neorizobium sp. ( Neorhizobium sp.) FWR7 (KACC 81196BP) and Microbacterium testaceum ( Microbacterium testaceum ) FWT1 (KACC 81195BP) are preferably prepared by further mixing one or more microorganisms selected from among them.
한편, 상기 본 발명의 퇴비에 있어서, 상기 식품 관련 폐기물은 바람직하게 음식물 폐기물을 4~6, 커피박을 4~6, 왕겨를 0.8~1.2의 무게 비율로 포함하는 것이 좋다.Meanwhile, in the compost of the present invention, the food-related waste preferably contains food waste in a weight ratio of 4 to 6, coffee grounds in a weight ratio of 4 to 6, and rice husk in a weight ratio of 0.8 to 1.2.
본 발명의 아조스피릴룸 sp. (Azospirillum sp.) FWR9 (KACC 81197BP)는 고분자의 유기성 물질 (탄수화물, 단백질 및 지방 등)을 적절한 크기로 분해하여, 작물이 흡수하기 좋게 변환시키는 특징이 있으며, 우수한 질소 고정화능력을 가지고 있다. 따라서, 본 발명의 아조스피릴룸 sp. (Azospirillum sp.) FWR9 (KACC 81197BP)를 이용한 퇴비는 작물이 잘 자라게 된다.Azospirillum sp of the present invention. ( Azospirillum sp.) FWR9 (KACC 81197BP) has the characteristic of breaking down high molecular weight organic substances (carbohydrates, proteins, fats, etc.) into appropriate sizes, converting them to be easily absorbed by crops, and has excellent nitrogen fixation ability. Therefore, Azospirillum sp of the present invention. ( Azospirillum sp.) Compost using FWR9 (KACC 81197BP) allows crops to grow well.
도 1은 본 발명 균주들을 분리하기 위해, 음식물 퇴비화 장치 내에서 시료를 수득한 후, 플레이트에 배양한 것을 보여준다.Figure 1 shows that in order to isolate strains of the present invention, samples were obtained from a food composting device and then cultured on a plate.
도 2는 본 발명 네오리조비움 sp. (Neorhizobium sp.) FWR7 (KACC 81196BP)의 계통도 분석 결과이다.Figure 2 shows the present invention Neorizobium sp. ( Neorhizobium sp.) This is the result of phylogenetic analysis of FWR7 (KACC 81196BP).
도 3은 본 발명 아조스피릴룸 sp. (Azospirillum sp.) FWR9 (KACC 81197BP)의 계통도 분석 결과이다.Figure 3 shows the present invention Azospirillum sp. ( Azospirillum sp.) This is the result of phylogenetic analysis of FWR9 (KACC 81197BP).
도 4는 본 발명 마이크로박테리움 테스타시움 (Microbacterium testaceum) FWT1 (KACC 81195BP)의 계통도 분석 결과이다.Figure 4 is a phylogenetic analysis result of Microbacterium testaceum FWT1 (KACC 81195BP) of the present invention.
도 5는 본 발명 네오리조비움 sp. (Neorhizobium sp.) FWR7 (KACC 81196BP), 아조스피릴룸 sp. (Azospirillum sp.) FWR9, 마이크로박테리움 테스타시움 (Microbacterium testaceum) FWT1의 현미경 관찰 결과이다.Figure 5 shows the present invention Neorizobium sp. ( Neorhizobium sp.) FWR7 (KACC 81196BP), Azospirillum sp. ( Azospirillum sp.) FWR9, Microbacterium testaceum FWT1 microscopic observation results.
도 6은 본 발명 네오리조비움 sp. (Neorhizobium sp.) FWR7 (KACC 81196BP)의 효소활성 및 생리특성 분석 결과이다.Figure 6 shows the present invention Neorizobium sp. ( Neorhizobium sp.) This is the result of analysis of enzyme activity and physiological characteristics of FWR7 (KACC 81196BP).
도 7은 본 발명 아조스피릴룸 sp. (Azospirillum sp.) FWR9 (KACC 81197BP)의 효소활성 및 생리특성 분석 결과이다.Figure 7 shows the present invention Azospirillum sp. ( Azospirillum sp.) This is the result of analysis of enzyme activity and physiological characteristics of FWR9 (KACC 81197BP).
도 8은 본 발명 마이크로박테리움 테스타시움 (Microbacterium testaceum) FWT1 (KACC 81195BP)의 효소활성 및 생리특성 분석 결과이다.Figure 8 shows the results of analysis of enzyme activity and physiological characteristics of Microbacterium testaceum FWT1 (KACC 81195BP) of the present invention.
도 9는 커피박, 음식물 폐기물 및 왕겨를 혼합한 후, 2일간 발효시켜 얻은 1차 발효 퇴비를 일반 상토와 1:1의 중량비율로 혼합한 후, 30일간 상추를 재배하며, 1차 발효 퇴비의 작물 생장 촉진능을 일반 상토와 비교한 결과이다.Figure 9 shows the primary fermentation compost obtained by mixing coffee waste, food waste, and rice husk and fermenting for 2 days, mixing it with general topsoil in a weight ratio of 1:1, then cultivating lettuce for 30 days, and primary fermentation compost. This is the result of comparing the crop growth promotion ability of with general soil.
도 10은 상기 1차 발효 퇴비에, 본 발명 네오리조비움 sp. (Neorhizobium sp.) FWR7 (KACC 81196BP)를 포함하는 동결건조분말 (혼합균주)을 첨가하고, 30일간 발효시켜 2차 발효 퇴비를 제조하고, 상기 2차 발효 퇴비를 일반 상토와 1:1, 2:1 중량비율로 혼합한 후, 30일간 상추를 재배하며, 2차 발효 퇴비의 작물 생장 촉진능을 일반 상토와 비교한 결과이다.Figure 10 shows the primary fermentation compost, Neorizobium sp. of the present invention. ( Neorhizobium sp.) Freeze-dried powder (mixed strains) containing FWR7 (KACC 81196BP) was added and fermented for 30 days to prepare secondary fermentation compost, and the secondary fermentation compost was mixed with general topsoil 1:1, 2 :1 This is the result of growing lettuce for 30 days after mixing at a weight ratio, and comparing the crop growth promotion ability of secondary fermentation compost with general topsoil.
본 발명은 아조스피릴룸 sp. (Azospirillum sp.) FWR9 (KACC 81197BP)를 제공한다. The present invention relates to Azospirillum sp. ( Azospirillum sp.) FWR9 (KACC 81197BP) is provided.
상기 본 발명의 아조스피릴룸 sp. (Azospirillum sp.) FWR9 (KACC 81197BP)는 음식물 폐기물 퇴비화 장치에서 분리해 낸 것으로, 서열번호 2의 16S rRNA 서열을 갖는다. 하기 실시예에서, 서열번호 2로 유전적 동정한 결과, 가장 유사한 균주와 오직 98.1% 만이 유사하였다. 이를 통해 본 발명의 균주는 아조스피릴룸 sp. (Azospirillum sp.)의 신규 균주임을 확인할 수 있었다.Azospirillum sp of the present invention. ( Azospirillum sp.) FWR9 (KACC 81197BP) was isolated from a food waste composting device and has the 16S rRNA sequence of SEQ ID NO: 2. In the following examples, as a result of genetic identification using SEQ ID NO: 2, only 98.1% of the strain was similar to the most similar strain. Through this, the strain of the present invention is Azospirillum sp. It was confirmed that it was a new strain of ( Azospirillum sp.).
한편, 본 발명의 아조스피릴룸 sp. (Azospirillum sp.) FWR9 (KACC 81197BP)는 작물 재배에 유용한 퇴비 제조에 사용하기 좋다. 본 발명의 아조스피릴룸 sp. (Azospirillum sp.) FWR9 (KACC 81197BP)는 다양한 효소활성 및 생리특성을 보이는데, 퇴비에 포함되어 있는 고분자 물질들을 저분자 물질로 분해하여 작물들이 퇴비에 포함된 유용성분 흡수를 더욱 쉽게 할 수 있도록 한다. 또한, 질소순환에 관여하여, 작물 생장에 도움을 줄 수 있다. 하기 실시예에 따르면, 본 발명의 아조스피릴룸 sp. (Azospirillum sp.) FWR9 (KACC 81197BP)를 사용하여 발효시킨 퇴비의 작물 생장 촉진능이 우수하다.On the other hand, Azospirillum sp of the present invention. ( Azospirillum sp.) FWR9 (KACC 81197BP) is good for use in producing compost useful for crop cultivation. Azospirillum sp of the present invention. ( Azospirillum sp.) FWR9 (KACC 81197BP) exhibits various enzyme activities and physiological characteristics. It decomposes the high molecular substances contained in compost into low molecular substances, allowing crops to more easily absorb useful substances contained in the compost. Additionally, it is involved in the nitrogen cycle and can help crop growth. According to the following examples, Azospirillum sp. of the present invention. ( Azospirillum sp.) Compost fermented using FWR9 (KACC 81197BP) has excellent crop growth promotion ability.
또한, 본 발명은 아조스피릴룸 sp. (Azospirillum sp.) FWR9 (KACC 81197BP)를 이용한 식품 관련 폐기물의 퇴비화를 위한 미생물 제제를 제공한다.In addition, the present invention is Azospirillum sp. A microbial preparation for composting of food-related waste using ( Azospirillum sp.) FWR9 (KACC 81197BP) is provided.
상기 미생물 제제는 본 발명의 아조스피릴룸 sp. (Azospirillum sp.) FWR9 (KACC 81197BP)를 포함하는 것을 의미하는데, 일예로는 균주 건조분말 형태로 제조된 것을 들 수 있다. 이렇게 제조된 미생물 제제를 퇴비 제조 장치에 투입하는 방식으로 사용할 수 있다.The microbial agent is Azospirillum sp. of the present invention. ( Azospirillum sp.) This means that it contains FWR9 (KACC 81197BP), and as an example, the strain may be manufactured in the form of dried powder. The microbial preparation prepared in this way can be used by injecting it into a compost manufacturing device.
한편, 본 발명에서 상기 식품 관련 폐기물은 도축장의 도축 폐기물, 식품 가공 공장의 폐기물, 가정에서 배출되는 음식물 쓰래기 등의 폐기물을 의미한다.Meanwhile, in the present invention, the food-related waste refers to waste such as slaughter waste from a slaughterhouse, waste from a food processing plant, and food waste discharged from homes.
또한, 본 발명은 아조스피릴룸 sp. (Azospirillum sp.) FWR9 (KACC 81197BP) 및 식품 관련 폐기물을 혼합하여 제조한 것을 특징으로 하는 퇴비를 제공한다. In addition, the present invention is Azospirillum sp. ( Azospirillum sp.) Provides compost that is manufactured by mixing FWR9 (KACC 81197BP) and food-related waste.
또한, 본 발명은 네오리조비움 sp. (Neorhizobium sp.) FWR7 (KACC 81196BP), 마이크로박테리움 테스타시움 (Microbacterium testaceum) FWT1 (KACC 81195BP) 중 선택되는 어느 하나 이상의 미생물과 아조스피릴룸 sp. (Azospirillum sp.) FWR9 (KACC 81197BP) 및 식품 관련 폐기물을 혼합하여 제조한 것을 특징으로 하는 퇴비를 제공한다.In addition, the present invention is Neorizobium sp. ( Neorhizobium sp.) FWR7 (KACC 81196BP), one or more microorganisms selected from Microbacterium testaceum FWT1 (KACC 81195BP) and Azospirillum sp. ( Azospirillum sp.) Provides compost that is manufactured by mixing FWR9 (KACC 81197BP) and food-related waste.
상기 FWR9, FWR7 및 FWT1는 식품 관련 폐기물을 작게 분해하여, 작물 등이 퇴비에 포함된 유기성 물질들을 더욱 쉽게 흡수할 수 있도록 도와준다. 따라서, 상기와 같이 제조된 퇴비를 작물 재배에 사용하는 경우 작물이 잘 자라게 된다. 한편, 상기 FWR9, FWR7 및 FWT1는 바람직하게 각각 0.8~1.2:0.8~1.2:0.8~1.2의 비율로 혼합하여 사용하는 것이 좋다.The FWR9, FWR7, and FWT1 break down food-related waste into small pieces, helping crops and other plants more easily absorb organic substances contained in compost. Therefore, when the compost prepared as above is used for crop cultivation, the crops grow well. Meanwhile, FWR9, FWR7 and FWT1 are preferably mixed and used in a ratio of 0.8 to 1.2:0.8 to 1.2:0.8 to 1.2, respectively.
한편, 퇴비 제조시에 상기 식품 관련 폐기물로, 바람직하게, 음식물 폐기물, 커피박 및 왕겨를 혼합한 것을 사용하는 것이 좋다.Meanwhile, when producing compost, it is advisable to use a mixture of food waste, coffee grounds, and rice husk as the food-related waste.
매립 등으로 폐기되는 음식물 폐기물, 커피박, 왕겨를 더욱 활용하여 자원 순환에 도움을 줄 수 있다. 특히, 커피박의 경우 주로, 커피 매장에서 발생되는 생활 폐기물로, 재활용율이 0.1% 정도로 매우 낮은 것으로 알려져 있다.Food waste, coffee waste, and rice husk that are discarded in landfills can be further utilized to help resource circulation. In particular, coffee grounds are mainly household waste generated at coffee shops, and the recycling rate is known to be very low, around 0.1%.
게다가, 하기 실시예에 따르면, 음식물 폐기물을 4~6, 커피박을 4~6, 왕겨를 0.8~1.2의 무게 비율로 포함하도록 첨가하여 제조하는 경우, 퇴비 성분 규격에 적합하면서도, 질소, 인산, 칼리 함량이 높아 작물 재배에 유용한 퇴비를 제조할 수 있다. In addition, according to the following examples, when manufactured by adding food waste at a weight ratio of 4 to 6, coffee grounds at 4 to 6, and rice husk at a weight ratio of 0.8 to 1.2, it conforms to the compost component specifications and contains nitrogen, phosphoric acid, and It has a high potassium content, making it possible to produce compost useful for growing crops.
한편, 본 발명의 퇴비는 제조시에 기타 보충성분들을 더욱 첨가하여 제조된 것일 수 있다. 즉, 더욱 다양한 성분들을 갖게 하거나, 기존 성분들을 보충하여 재배하고자 하는 작물 특징에 더욱 맞는 퇴비를 제조할 수 있는 것이다. 이때, 사용하는 기타 보충성분 또한 발명의 취지에 맞추어 식품 관련 폐기물을 사용하는 것이 좋은데, 바람직한 예시로 골분, 술지게미, 깻묵을 들 수 있다.On the other hand, the compost of the present invention may be manufactured by adding other supplementary ingredients during production. In other words, it is possible to manufacture compost that is more suited to the characteristics of the crop to be grown by adding more diverse ingredients or supplementing existing ingredients. At this time, it is better to use food-related waste as other supplementary ingredients in accordance with the purpose of the invention. Preferred examples include bone meal, sake lees, and seed cake.
이하, 본 발명의 내용을 하기 실시예를 통하여 보다 상세하게 설명하고자 한다. 다만, 본 발명의 권리범위가 하기 실시예에만 한정되는 것은 아니고 그와 등가의 기술적 사상의 변형까지를 포함한다.Hereinafter, the contents of the present invention will be described in more detail through the following examples. However, the scope of the present invention is not limited to the following examples and includes modifications of the technical idea equivalent thereto.
[실시예 1: 균주 분리 및 동정][Example 1: Strain isolation and identification]
1) 균주 분리1) Strain isolation
음식물 처리 장치에 사용하기 적합한 미생물을 분리하고자 했다. 이를 위해 2년동안 안정적으로 운용되던 음식물 퇴비화 장치 내에서 시료를 채취한 후, 희석하고, 도 1 처럼 플레이트에 배양하여 총 43 종의 균주를 분리할 수 있었다. We sought to isolate microorganisms suitable for use in food processing devices. For this purpose, samples were collected from a food composting device that had been operating stably for two years, diluted, and cultured on plates as shown in Figure 1, allowing a total of 43 strains to be isolated.
2) 균주 동정2) Strain identification
43 종의 균주에 대하여, 식품 관련 폐기물의 퇴비화 관련 실험 진행하고, 퇴비제조에 적합한 균주를 선택하였다. 이를 통해, FWR9, FWT1, FWR7가 균주가 식품 관련 폐기물의 퇴비화에 우수한 것을 확인할 수 있었다.Experiments related to composting of food-related waste were conducted on 43 strains, and strains suitable for compost production were selected. Through this, it was confirmed that strains FWR9, FWT1, and FWR7 were excellent at composting food-related waste.
이후 FWR9, FWR7, FWT1 균주들을 바이오팩트회사에 의뢰하여 16S rRNA 분석을 진행하였다. Afterwards, FWR9, FWR7, and FWT1 strains were requested from Biofact Company and 16S rRNA analysis was performed.
16S rRNA 분석한 결과, FWR7 균주의 경우, 서열번호 1의 16S rRNA 서열을 얻을 수 있었다. 서열번호 1로 유전적 동정한 결과 네오리조비움 토메질렌세 (Neorhizobium tomejilense)와 서열이 약 97.3% 유사한 것을 확인할 수 있었다. 또한, 계통도 분석 (도 2)을 통해, 기존 속내의 다른 종과 구별되는 신규의 균주임을 확인할 수 있었다. 또한, FWR7 균주의 멤브레인의 지방산 특성을 분석한 결과 (표 1)에서도 기존 알려진 균주와 유사도 지수가 낮게 나오는 것을 확인할 수 있었다. 이를 바탕으로 본 발명의 FWR7 균주는 신규의 네오리조비움 sp. (Neorhizobium sp.) 종류의 균주임을 알 수 있었다. As a result of 16S rRNA analysis, in the case of strain FWR7, the 16S rRNA sequence of SEQ ID NO: 1 was obtained. As a result of genetic identification using sequence number 1, it was confirmed that the sequence was approximately 97.3% similar to Neorhizobium tomejilense . In addition, through phylogenetic analysis (Figure 2), it was confirmed that it was a new strain distinct from other species within the existing genus. In addition, as a result of analyzing the fatty acid characteristics of the membrane of the FWR7 strain (Table 1), it was confirmed that the similarity index was low compared to the previously known strains. Based on this, the FWR7 strain of the present invention is a novel Neorizobium sp. It was found that it was a type of strain ( Neorhizobium sp.).
| LibraryLibrary | 유사도 지수(Sim index)Similarity index (Sim index) | 균주strain |
| RTSBA6 6.10RTSBA6 6.10 | 0.5140.514 |
주글로에아-라미제라 (Zoogloea-ramigera)Zoogloea-lamigera ( Zoogloea-ramigera ) |
| 0.4890.489 |
로즈오모나스-파우리아 (Roseomonas-fauriae)Rose Omonas-Pauria ( Rosomonas-fauriae ) |
|
| 0.3740.374 |
리조비움-라디오박테르 (Rhizobium-radiobacter)Rhizobium-Radiobacter ( Rhizobium-radiobacter ) |
|
| 0.3440.344 |
글루코박터-아사이/세리누스/옥시단스 (Gluconobacter-asaii/cerinus/oxydans)Glucobacter-Acai/Serinus/Oxidans ( Gluconobacter-asaii/cerinus/oxydans ) |
FWR9 균주의 경우, 서열번호 2의 16S rRNA 서열을 얻을 수 있었다. 서열번호 2로 유전적 동정한 결과 아조스피릴룸 티오필룸 (Azospirillum thiophilum)과 서열이 약 98.1% 유사한 것을 확인할 수 있었다. 또한, 계통도 분석 (도 3)을 통해, 기존 속내의 다른 종과 구별되는 것을 확인할 수 있었다. 또한, FWR9 균주의 멤브레인의 지방산 특성을 분석한 결과 (표 2)에서도 기존 알려진 균주와 유사도 지수가 낮게 나오는 것을 확인할 수 있었다. 이를 바탕으로 본 발명의 FWR9 균주는 신규의 아조스피릴룸 sp. (Azospirillum sp.) 종류의 균주임을 알 수 있었다.For the FWR9 strain, the 16S rRNA sequence of SEQ ID NO: 2 was obtained. As a result of genetic identification using SEQ ID No. 2, it was confirmed that the sequence was approximately 98.1% similar to Azospirillum thiophilum . In addition, through phylogenetic analysis (Figure 3), it was confirmed that it was distinguished from other species within the existing genus. In addition, as a result of analyzing the fatty acid characteristics of the membrane of the FWR9 strain (Table 2), it was confirmed that the similarity index was low compared to the previously known strains. Based on this, the FWR9 strain of the present invention is a new Azospirillum sp. ( Azospirillum sp.) It was found to be a type of strain.
| LibraryLibrary | 유사도 지수(Sim index)Similarity index (Sim index) | 균주strain |
| RTSBA6 6.10RTSBA6 6.10 | 0.4730.473 |
아조스피릴룸 브라질렌세 (Azospirillum brasilense)Azospirillum brasilense ( Azospirillum brasilense ) |
| 0.4200.420 |
로조모나스 파우리아 (Roseomonas fauriae)Rozomonas pauria ( Roseomonas fauriae ) |
FWT1 균주의 경우, 서열번호 3의 16S rRNA 서열을 얻을 수 있었다. 서열번호 3으로 유전적 동정한 결과 마이크로박테리움 테스타시움 (Microbacterium testaceum)과 서열이 약 99.13% 유사한 것을 확인할 수 있었다. 또한, 계통도 분석 (도 4)을 통해, 마이크로박테리움 테스타시움 (Microbacterium testaceum) 균주임을 알 수 있었다. For the FWT1 strain, the 16S rRNA sequence of SEQ ID NO: 3 was obtained. As a result of genetic identification using SEQ ID NO: 3, it was confirmed that the sequence was approximately 99.13% similar to Microbacterium testaceum . In addition, through phylogenetic analysis (FIG. 4), it was found to be a Microbacterium testaceum strain.
한편, 상기 FWR7, FWR9, FWT1 균주를 각각 농촌진흥청 국립농업과학원에 기탁하고 수탁번호를 부여받아, 최종적으로 네오리조비움 sp. (Neorhizobium sp.) FWR7 (KACC 81196BP), 마이크로박테리움 테스타시움 (Microbacterium testaceum) FWT1 (KACC 81195BP), 아조스피릴룸 sp. (Azospirillum sp.) FWR9 (KACC 81197BP)으로 명명하였다.Meanwhile, the FWR7, FWR9, and FWT1 strains were each deposited at the National Institute of Agricultural Science of the Rural Development Administration and given accession numbers, and finally, Neorizobium sp. ( Neorhizobium sp.) FWR7 (KACC 81196BP), Microbacterium testaceum FWT1 (KACC 81195BP), Azospirillum sp. ( Azospirillum sp.) was named FWR9 (KACC 81197BP).
[실시예 2: 규주 특징 확인 실험][Example 2: Experiment to confirm the characteristics of silica]
1) 배양 및 현미경 관찰1) Culture and microscopic observation
본 발명의 네오리조비움 sp. (Neorhizobium sp.) FWR7을 배양한 결과, 적합 배양온도는 20~37℃, 최적 배양온도는 25℃, 적합 배양염도는 0~2.0%, 적합 배양pH는 7~8인 것을 확인할 수 있었다. Neorizobium sp of the present invention. ( Neorhizobium sp.) As a result of cultivating FWR7, it was confirmed that the suitable culture temperature was 20~37℃, the optimal culture temperature was 25℃, the suitable culture salinity was 0~2.0%, and the suitable culture pH was 7~8.
본 발명의 아조스피릴룸 sp. (Azospirillum sp.) FWR9를 배양한 결과, 적합 배양온도는 20~37℃, 최적 배양온도는 25℃, 적합 배양염도는 0~1.5%, 적합 배양pH는 7~8인 것을 확인할 수 있었다. Azospirillum sp of the present invention. ( Azospirillum sp.) As a result of cultivating FWR9, it was confirmed that the suitable culture temperature was 20~37℃, the optimal culture temperature was 25℃, the suitable culture salinity was 0~1.5%, and the suitable culture pH was 7~8.
본 발명의 마이크로박테리움 테스타시움 (Microbacterium testaceum) FWT1을 배양한 결과, 적합 배양온도는 20~37℃, 최적 배양온도는 25℃, 적합 배양염도는 0~2.0%, 적합 배양 pH는 7~8인 것을 확인할 수 있었다. As a result of cultivating Microbacterium testaceum FWT1 of the present invention, the suitable culture temperature was 20 to 37 ℃, the optimal culture temperature was 25 ℃, the suitable culture salinity was 0 to 2.0%, and the suitable culture pH was 7 to 7. I was able to confirm that it was 8.
현미경으로 관찰한 결과, FWR7, FWR9 및 FWT1는 모두 간균 형태의 균주임을 확인할 수 있었다 (도 5). As a result of observation under a microscope, it was confirmed that FWR7, FWR9, and FWT1 were all bacillus-type strains (FIG. 5).
2) FWR7의 효소활성 및 생리특성 측정2) Measurement of enzyme activity and physiological characteristics of FWR7
네오리조비움 sp. (Neorhizobium sp.) FWR7의 효소활성을 테스트한 결과, 알칼리성 인산분해효소 (Alkaline phosphatase), 에스트레이즈(Estrease, C4), 에스트레이즈 리파아제 (Estrease Lipase, CB), 류신 아릴아미다제 (Leucine arylamidase), 트립신(Trypsin), 알파-키모트립신(α-chymotrypsin), 산성 포스파타제 (Acid phospatase), 나프톨-AS-Bl-포스포하이드롤라제(Naphtol-AS-Bl-phosphohydrolase), 베타-글루쿠로니다제(β-glucurorndase), 알파-글루코시다제(α-glucosidase), 베타-글루코시다아제(β-glucosidase), N-아세틸-베타-글루코사미닌다제(N-acetyl-β－glucosaminindase)가 양성으로 나타났다 (도 6의 a).Neorizobium sp. ( Neorhizobium sp.) As a result of testing the enzyme activity of FWR7, alkaline phosphatase, Estrease (C4), Estrease Lipase (CB), Leucine arylamidase, Trypsin, α-chymotrypsin, acid phospatase, Naphtol-AS-Bl-phosphohydrolase, beta-glucuronidase (β-glucurorndase), alpha-glucosidase, beta-glucosidase, and N-acetyl-beta-glucosaminindase were positive. appeared (a in Figure 6).
네오리조비움 sp. (Neorhizobium sp.) FWR7의 생리특성을 테스트한 결과, 아르기닌 이가수분해 (arginine dihydrolase), 우레아제(urease), hydrolysis (b-glucosidase), 베타-글루코시다제(β-glucosidase), 포도당 동화 (glucose assimilation), 아라비노스 동화 (arabinose assimilation), 만노스 동화 (mannose assimilation), 만니톨 동화 (mannitol assimilation), N-아세틸-글루코사민 동화 (N-acetyl-glucosamine assimilation), 말토스 동화 (maltose assimilation), 글루코네이트 동화 (gluconate assimilation), 말산 동화 (malate assimilation)가 양성으로 나타났다 (도 6의 b).Neorizobium sp. ( Neorhizobium sp.) As a result of testing the physiological properties of FWR7, the results showed arginine dihydrolase, urease, hydrolysis (b-glucosidase), beta-glucosidase, and glucose assimilation. assimilation, arabinose assimilation, mannose assimilation, mannitol assimilation, N-acetyl-glucosamine assimilation, maltose assimilation, gluconate Gluconate assimilation and malate assimilation were positive (Figure 6b).
한편, 상기 네오리조비움 sp. (Neorhizobium sp.) FWR7의 효소활성 측정 결과 및 생리특성 측정 결과는, 본 발명의 네오리조비움 sp. (Neorhizobium sp.) FWR7을 이용하여 퇴비를 제조하는 경우, 고분자 물질 (탄수화물, 지방, 단백질)을 저분자 물질로 바꾸어 작물 생장에 도움을 줄 수 있음을 의미한다. Meanwhile, the Neorizobium sp. ( Neorhizobium sp.) The results of measuring enzyme activity and physiological characteristics of FWR7 are Neorhizobium sp. of the present invention. ( Neorhizobium sp.) This means that when producing compost using FWR7, high molecular substances (carbohydrates, fats, proteins) can be converted into low molecular substances to help crop growth.
3) FWR9의 효소활성 및 생리특성 측정3) Measurement of enzyme activity and physiological characteristics of FWR9
아조스피릴룸 sp. (Azospirillum sp.) FWR9의 효소활성을 테스트한 결과, 알칼리성 인산분해효소 (Alkaline phosphatase), 에스트레이즈(Estrease, C4), 에스트레이즈 리파아제 (Estrease Lipase, CB), 류신 아릴아미다제 (Leucine arylamidase), 트립신(Trypsin), 산성 포스파타제 (Acid phospatase), 나프톨-AS-Bl-포스포하이드롤라제(Naphtol-AS-Bl-phosphohydrolase), 베타-글루코시다아제(β-glucosidase), 알파-후코시다제(α-fucosidase)가 양성으로 나타났다 (도 7의 a).Azospirillum sp. ( Azospirillum sp.) As a result of testing the enzyme activity of FWR9, alkaline phosphatase, Estrease (C4), Estrease Lipase (CB), Leucine arylamidase, Trypsin, acid phospatase, Naphtol-AS-Bl-phosphohydrolase, beta-glucosidase, alpha-fucosidase ( α-fucosidase) was found to be positive (Figure 7a).
아조스피릴룸 sp. (Azospirillum sp.) FWR9의 생리특성을 테스트한 결과, 질산염의 감소 (duction of nitrates), 아르기닌 이가수분해 (arginine dihydrolase), 우레아제(urease), 베타-글루코시다제(β-glucosidase), 베타-갈락토시다제(β-galactosidase), 포도당 동화 (glucose assimilation), 만니톨 동화 (mannitol assimilation), N-아세틸-글루코사민 동화 (N-acetyl-glucosamine assimilation)가 양성으로 나타났다 (도 7의 b).Azospirillum sp. ( Azospirillum sp.) As a result of testing the physiological properties of FWR9, it was found that duction of nitrates, arginine dihydrolase, urease, β-glucosidase, beta- Galactosidase, glucose assimilation, mannitol assimilation, and N-acetyl-glucosamine assimilation were positive (Figure 7b).
한편, 상기 네오리조비움 아조스피릴룸 sp. (Azospirillum sp.) FWR9의 효소활성 측정 결과 및 생리특성 측정 결과는, 본 발명의 아조스피릴룸 sp. (Azospirillum sp.) FWR9를 이용하여 퇴비를 제조하는 경우, 고분자 물질 (탄수화물, 지방, 단백질)을 저분자 물질로 바꾸어 작물 생장에 도움을 줄 수 있음을 의미한다. 또한, 질소순환에 관여하여, 작물 생장에 도움을 줄 수 있음을 의미한다. Meanwhile, the Neorizobium azospirillum sp. ( Azospirillum sp.) The enzyme activity measurement results and physiological characteristics measurement results of FWR9 are the Azospirillum sp. of the present invention. ( Azospirillum sp.) This means that when producing compost using FWR9, high molecular substances (carbohydrates, fats, proteins) can be converted into low molecular substances to help crop growth. Additionally, it is involved in the nitrogen cycle, meaning it can help crop growth.
4) FWT1의 효소활성 및 생리특성 측정4) Measurement of enzyme activity and physiological characteristics of FWT1
마이크로박테리움 테스타시움 (Microbacterium testaceum) FWT1의 효소활성을 테스트한 결과, 에스트레이즈(Estrease, C4), 에스트레이즈 리파아제 (Estrease Lipase, CB), 리파제(Lipase, C14), 류신 아릴아미다제 (Leucine arylamidase), 발린 아릴아미다제 (Valine arylamidase), 크리스틴 아릴아미다제 (Crystine arylamidase), 트립신(Trypsin), 알파-키모트립신(α-chymotrypsin), 산성 포스파타제 (Acid phospatase), 나프톨-AS-Bl-포스포하이드롤라제(Naphtol-AS-Bl-phosphohydrolase), 베타-글루쿠로니다제(β-glucurorndase), 알파-글루코시다제(α-glucosidase), 베타-글루코시다아제(β-glucosidase), 알파-만노시다아제(α-mannosidase)가 양성으로 나타났다 (도 8의 a). As a result of testing the enzyme activity of Microbacterium testaceum FWT1, Estrease (C4), Estrease Lipase (CB), Lipase (C14), and Leucine Arylamidase arylamidase), Valine arylamidase, Crystine arylamidase, Trypsin, alpha-chymotrypsin, acid phosphatase, naphthol-AS-Bl-phos Naphtol-AS-Bl-phosphohydrolase, beta-glucurorndase, alpha-glucosidase, beta-glucosidase, alpha -Mannosidase (α-mannosidase) was found to be positive (Figure 8a).
마이크로박테리움 테스타시움 (Microbacterium testaceum) FWT1의 생리특성을 테스트한 결과, 아르기닌 이가수분해 (arginine dihydrolase), 우레아제(urease), 베타-갈락토시다제(β-galactosidase), 베타-갈락토시다제(β-galactosidase), 포도당 동화 (glucose assimilation), 아라비노스 동화 (arabinose assimilation), 만노스 동화 (mannose assimilation), 만니톨 동화 (mannitol assimilation), 말토스 동화 (maltose assimilation), 글루코네이트 동화 (gluconate assimilation), 말산 동화 (malate assimilation), 시트르산 동화 (citrate assimilation)가 양성으로 나타났다 (도 8의 b).As a result of testing the physiological properties of Microbacterium testaceum FWT1, arginine dihydrolase, urease, beta-galactosidase, beta-galactosidase β-galactosidase, glucose assimilation, arabinose assimilation, mannose assimilation, mannitol assimilation, maltose assimilation, gluconate assimilation ), malate assimilation, and citrate assimilation were found to be positive (Figure 8b).
한편, 상기 마이크로박테리움 테스타시움 (Microbacterium testaceum) FWT1의 효소활성 측정 결과 및 생리특성 측정 결과는, 본 발명의 마이크로박테리움 테스타시움 (Microbacterium testaceum) FWT1을 이용하여 퇴비를 제조하는 경우, 고분자 물질 (탄수화물, 지방, 단백질)을 저분자 물질로 바꾸어 작물 생장에 도움을 줄 수 있음을 의미한다. On the other hand, the enzyme activity measurement results and physiological characteristics measurement results of Microbacterium testaceum FWT1 are, when producing compost using Microbacterium testaceum FWT1 of the present invention, the polymer This means that substances (carbohydrates, fats, proteins) can be converted into low-molecular substances to help crops grow.
[실시예 3: 본 발명의 균주를 이용한 퇴비 제조 및 퇴비의 특성 확인 실험][Example 3: Compost production and compost characteristics confirmation experiment using the strain of the present invention]
본 실시예에서는 본 발명의 균주를 이용한 퇴비가 식물 재배에 유용한 특징을 보일 수 있는지, 확인하고자 했다. 또한, 어떠한 구성으로 퇴비를 제조해야, 식품 관련 폐기물 처리에 유용하면서도, 식물 재배에 더욱 유용한 특징을 보일 수 있는지 확인하고자 했다. In this example, we sought to confirm whether compost using the strain of the present invention could show useful characteristics for plant cultivation. In addition, we wanted to determine what composition compost should be made in so that it can be useful for processing food-related waste and show more useful characteristics for plant cultivation.
3-1) 퇴비 제조를 위한 음식물 폐기물 회득3-1) Harvesting food waste for compost production
본 발명에서 사용하는 음식물 폐기물은 음식물 퇴비화 장치에서 얻을 수 있었다. 구체적으로, 상기 음식물 퇴비화 장치는 희석발효방식으로 음식물을 처리하는데, 음식물을 발효시켜 감량시키고, 탈수시킨 후, 남은 음식물 찌꺼기를 회수장치에 모아두는 장치이다. 본 발명에서는 상기 음식물 퇴비화 장치에 음식물을 투입한 후, 2일이 지난 시점에, 상기 회수장치에 모인 음식물 찌꺼기를 퇴비 제조를 위한 음식물 폐기물로 사용하였다.The food waste used in the present invention was obtained from a food composting device. Specifically, the food composting device processes food using a dilution fermentation method. It is a device that ferments food to reduce its weight, dehydrates it, and then collects the remaining food waste in a recovery device. In the present invention, two days after putting food into the food composting device, the food scraps collected in the recovery device were used as food waste for compost production.
3-2) 음식물 폐기물 및 커피박을 이용한 1차 발효 퇴비 제조3-2) Production of primary fermentation compost using food waste and coffee grounds
음식물 폐기물 및 커피박을 이용한 퇴비는 상기 회수장치 내에서 제조할 수 있었다. 구체적으로 음식물 폐기물이 있는 회수장치에, 커피박 및 왕겨를 투입한 후 2일간 발효시켜 제조하였다. 구체적인 첨가 비율은 하기 표 3에 나타냈다. Compost using food waste and coffee grounds could be produced within the recovery device. Specifically, it was manufactured by putting coffee grounds and rice husk into a recovery device containing food waste and fermenting it for 2 days. The specific addition ratio is shown in Table 3 below.
| 명칭designation | 커피박(kg)Coffee grounds (kg) | 음식물(kg)Food (kg) | 왕겨(kg)Rice husk (kg) |
| 제조예 1Manufacturing Example 1 | 55 | 2.52.5 | 2.52.5 |
| 제조예 2Production example 2 | 77 | 1.51.5 | 1.51.5 |
| 제조예 3Production example 3 | 33 | 3.53.5 | 3.53.5 |
| 제조예 4Production example 4 | 55 | 55 | -- |
| 제조예 5Production example 5 | 55 | 44 | 1One |
제조예 1 내지 3을 제조하며 관찰한 결과, 커피박:음식물의 중량비율이 1:1보다 높아져 음식물의 중량비율이 더 큰 비율을 차지하게 되면, 퇴비 발효 과정중에 악취가 발생하는 문제점을 확인할 수 있었다. 따라서, 이후 제조과정에서는 음식물의 중량이 커피박의 중량을 넘지 않도록 퇴비를 제조하였다.As a result of observation while manufacturing Preparation Examples 1 to 3, it can be seen that when the weight ratio of coffee waste:food becomes higher than 1:1 and the weight ratio of food takes up a larger proportion, the problem of bad smell occurring during the compost fermentation process can be confirmed. there was. Therefore, in the subsequent manufacturing process, compost was manufactured so that the weight of the food did not exceed the weight of the coffee grounds.
제조예 4를 제조한 후, 비토분석센타 주식회사에 의뢰하여, 퇴비 성분을 검사한 결과, 퇴비 성분 규격에 대부분 적합하였지만, 수분 함량이 다소 높게 나왔다. After preparing Preparation Example 4, Vito Analysis Center Co., Ltd. was requested to test the compost components, and as a result, most of them met the compost component specifications, but the moisture content was somewhat high.
제조예 5를 제조한 후, 퇴비 성분을 검사한 결과, 수분 함량이 적정하게 나와 퇴비로 사용하기에 적합하였다. 게다가, 제조예 4의 경우보다 인산 및 칼리 성분이 높아져 작물 재배에 더욱 적합하였다.After preparing Preparation Example 5, the compost ingredients were tested and the moisture content was found to be appropriate, making it suitable for use as compost. In addition, the phosphoric acid and potassium components were higher than those of Preparation Example 4, making it more suitable for crop cultivation.
3-3) 1차 발효 퇴비를 이용한 상추 재배 실험3-3) Lettuce cultivation experiment using primary fermentation compost
상기 실시예 3-2)에서 제조한 퇴비를 이용하여 상추를 재배하며, 일반 상토와 비교하였다. Lettuce was grown using the compost prepared in Example 3-2) and compared with general topsoil.
구체적으로, 일반 상토와 제조예 4, 제조예 5를 각각 1:1의 중량비율로 혼합한 후, 30일간 상추를 재배하고, 뿌리의 생장, 잎의 크기 등을 종합적으로 비교하였다 (도 9). Specifically, after mixing general topsoil with Preparation Example 4 and Preparation Example 5 at a weight ratio of 1:1, lettuce was grown for 30 days, and root growth, leaf size, etc. were comprehensively compared (FIG. 9) .
이를 통해 상기 제조한 1차 발효 퇴비가 작물 생장에 도움을 주는 것을 확인할 수 있었다. 게다가, 왕겨가 더욱 첨가된 퇴비의 경우, 작물이 더욱 잘 재배되는 것을 확인할 수 있었다.Through this, it was confirmed that the prepared primary fermentation compost helped crop growth. In addition, it was confirmed that crops grew better in the case of compost with more rice husk added.
3-4) 미생물이 더욱 첨가된 2차 발효 퇴비 제조3-4) Manufacturing secondary fermentation compost with additional microorganisms
1차 발효 퇴비에 미생물을 첨가하고 발효시킨 퇴비를 제조하였다. Microorganisms were added to the first fermentation compost and fermented compost was prepared.
구체적으로, 상기 3-2)에 기재한 방식으로 커피박 5kg, 음식물 폐기물 5kg, 왕겨 1kg이 혼합된 1차 발효 퇴비를 제조하였다. 제조된 1차 발효 퇴비에 혼합균주 0.1g을 혼합시킨 후, 30일간 2차 발효시켜, 2차 발효 퇴비를 제조하였다. Specifically, primary fermentation compost was prepared by mixing 5 kg of coffee grounds, 5 kg of food waste, and 1 kg of rice husk by the method described in 3-2) above. After mixing 0.1 g of mixed strains with the prepared primary fermentation compost, secondary fermentation was performed for 30 days to prepare secondary fermentation compost.
한편, 상기 사용된 혼합균주는 상기 실시예 1에서 기재한 본 발명의 네오리조비움 sp. (Neorhizobium sp.) FWR7, 아조스피릴룸 sp. (Azospirillum sp.) FWR9 및 마이크로박테리움 테스타시움 (Microbacterium testaceum) FWT1의 균주 배양액을 동결건조한 후, 1:1:1의 무게비율로 혼합하여 제조한 것으로, 1 x 1012 CFU/mg의 농도를 가진다. Meanwhile, the mixed strain used was Neorizobium sp. of the present invention described in Example 1. ( Neorhizobium sp.) FWR7, Azospirillum sp. ( Azospirillum sp.) FWR9 and Microbacterium testaceum ( Microbacterium testaceum) FWT1 strain cultures were freeze-dried and mixed at a weight ratio of 1:1:1 to produce a concentration of 1 x 10 12 CFU/mg. has
한편, 상기 2차 발효 과정을 관찰한 결과, 30일 동안의 2차 발효 후, 퇴비가 부엽토 냄새로 완전 숙성된 것을 확인할 수 있었다. 이후, 제조된 퇴비 성분을 검사하고, 상기 제조예 5의 퇴비 성분과 비교한 결과, 질소, 인산 및 칼리 성분을 더욱 많이 함유하는 것을 확인할 수 있었다. Meanwhile, as a result of observing the secondary fermentation process, it was confirmed that after secondary fermentation for 30 days, the compost was completely matured with a humus smell. Afterwards, the prepared compost components were inspected and compared with the compost components of Preparation Example 5, and it was confirmed that they contained more nitrogen, phosphoric acid, and potassium components.
3-5) 2차 발효 퇴비를 이용한 상추 재배 실험3-5) Lettuce cultivation experiment using secondary fermentation compost
상기 실시예 3-4)에서 제조한 퇴비를 이용하여 상추를 재배하며 일반 상토와 비교하였다. Lettuce was grown using the compost prepared in Example 3-4) and compared with general topsoil.
구체적으로, 상기 제조한 2차 발효 퇴비와 일반 상토를, 1:1, 2:1의 비율로 각각 혼합시킨 후, 30일간 상추를 재배하고 (도 10), 뿌리의 생장, 잎의 크기 등을 종합적으로 비교하였다. Specifically, the prepared secondary fermentation compost and general topsoil were mixed at a ratio of 1:1 and 2:1, respectively, and then lettuce was grown for 30 days (Figure 10), and root growth, leaf size, etc. Comprehensive comparison was made.
이를 통해, 2차 발효 퇴비의 비율이 높은 경우, 상추가 더욱 잘 재배되는 것을 확인할 수 있었다. 또한, 상기 3-3)의 제조예 5 (커피박, 음식물 폐기물, 왕겨)를 이용한 상추 재배 결과와 상기 2차 발표 퇴비 (커피박, 음식물 폐기물, 왕겨, 혼합균주)를 이용한 결과를 비교해도, 상추가 더욱 잘 자란 것을 확인할 수 있었다. Through this, it was confirmed that when the ratio of secondary fermentation compost was high, lettuce was grown better. In addition, when comparing the results of lettuce cultivation using Preparation Example 5 (coffee waste, food waste, rice husk) of 3-3) above and the results using the second published compost (coffee waste, food waste, rice husk, mixed strains), It was confirmed that the lettuce grew better.
이는 본 발명의 아조스피릴룸 sp. (Azospirillum sp.) FWR9 (KACC 81197BP)를 포함한 혼합균주가 퇴비 제조에 사용된 식품 관련 폐기물의 유기성 물질들을 적절한 크기로 분해하여, 작물이 흡수하기 좋게 변환하였음을 의미한다.This is Azospirillum sp. of the present invention. ( Azospirillum sp.) This means that the mixed strain including FWR9 (KACC 81197BP) decomposed the organic materials of food-related waste used in compost production into an appropriate size, converting them to an appropriate size for crops to absorb.
[규칙 제91조에 의한 정정 06.10.2023]
[Correction 06.10.2023 pursuant to Rule 91]
[규칙 제91조에 의한 정정 06.10.2023]
[Correction 06.10.2023 pursuant to Rule 91]
[규칙 제91조에 의한 정정 06.10.2023]
[Correction 06.10.2023 pursuant to Rule 91]
Claims (5)
- 트립신(Trypsin) 활성을 가지는 아조스피릴룸 sp. (Azospirillum sp.) FWR9 (KACC 81197BP). Azospirillum sp. with trypsin activity. ( Azospirillum sp.) FWR9 (KACC 81197BP).
- 트립신(Trypsin) 활성을 가지는 아조스피릴룸 sp. (Azospirillum sp.) FWR9 (KACC 81197BP)를 포함하는 것을 특징으로 하는 식품 관련 폐기물의 퇴비화를 위한 미생물 제제. Azospirillum sp. with trypsin activity. ( Azospirillum sp.) Microbial preparation for composting of food-related waste, characterized in that it contains FWR9 (KACC 81197BP).
- 트립신(Trypsin) 활성을 가지는 아조스피릴룸 sp. (Azospirillum sp.) FWR9 (KACC 81197BP) 및 식품 관련 폐기물을 혼합하여 제조한 것을 특징으로 하는 퇴비. Azospirillum sp. with trypsin activity. Compost produced by mixing ( Azospirillum sp.) FWR9 (KACC 81197BP) and food-related waste.
- 제3항에 있어서,According to paragraph 3,상기 퇴비는,The compost is,네오리조비움 sp. (Neorhizobium sp.) FWR7 (KACC 81196BP), 마이크로박테리움 테스타시움 (Microbacterium testaceum) FWT1 (KACC 81195BP) 중 선택되는 어느 하나 이상의 미생물을 더욱 혼합하여 제조한 것을 특징으로 하는 퇴비.Neorizobium sp. ( Neorhizobium sp.) FWR7 (KACC 81196BP), Microbacterium testaceum ( Microbacterium testaceum ) Compost characterized in that it is manufactured by further mixing one or more microorganisms selected from FWT1 (KACC 81195BP).
- 제3항 또는 제4항에 있어서,According to clause 3 or 4,상기 식품 관련 폐기물은,The food-related waste is:음식물 폐기물을 4~6, 커피박을 4~6, 왕겨를 0.8~1.2의 무게 비율로 포함하는 것임을 특징으로 하는 퇴비. Compost characterized in that it contains food waste in a weight ratio of 4 to 6, coffee grounds in a weight ratio of 4 to 6, and rice husk in a weight ratio of 0.8 to 1.2.
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| KR20200114265A (en) * | 2019-03-28 | 2020-10-07 | 김혜경 | Recycling method of organic waste and eco friendly-high functional Neo Humus Soil produced by the method |
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| KR101143902B1 (en) * | 2009-12-03 | 2012-05-10 | 라지수 | Manufacturing method of functional feedstuff of recycling food |
| KR101806156B1 (en) * | 2015-10-14 | 2017-12-07 | 라광수 | Method for manufacturing manure by microorganism using organic waste and the manure manufactured thereof |
| KR20200114265A (en) * | 2019-03-28 | 2020-10-07 | 김혜경 | Recycling method of organic waste and eco friendly-high functional Neo Humus Soil produced by the method |
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| NOVIA D, RAKHMADI A, PURWATI E, JULIYARSI I, HAIRANI R, SYALSAFILAH F: "The characteristics of organic fertilizer made of cow feces using the Indigenous Micro-Organisms (IMO) from raw manures", IOP CONFERENCE SERIES: EARTH AND ENVIRONMENTAL SCIENCE, vol. 287, no. 1, 1 July 2019 (2019-07-01), pages 012025, XP093146000, ISSN: 1755-1307, DOI: 10.1088/1755-1315/287/1/012025 * |
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