KR20190104353A - Bacillus megaliumium BC2-1 strain and food waste treatment method using the same - Google Patents
Bacillus megaliumium BC2-1 strain and food waste treatment method using the same Download PDFInfo
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- KR20190104353A KR20190104353A KR1020197021698A KR20197021698A KR20190104353A KR 20190104353 A KR20190104353 A KR 20190104353A KR 1020197021698 A KR1020197021698 A KR 1020197021698A KR 20197021698 A KR20197021698 A KR 20197021698A KR 20190104353 A KR20190104353 A KR 20190104353A
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- Prior art keywords
- bacillus
- kccm
- cereus
- megaterium
- food waste
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
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- C—CHEMISTRY; METALLURGY
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- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
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- Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)
Abstract
본 발명은 악취 제거 및 중금속 제거에 유용한 신균주 바실러스 메가테리움 BC2-1(Bacillus megaterium BC2-1) KCCM 10858P 및 이를 이용한 중금속, 악취제거, 음식물쓰레기를 이용한 액비제조방법 및 가축사료 생산방법에 관한 것이다. 본 발명에 따른 바실러스 메가터리움 BC2-1 KCCM 10858P는 축산폐수 및 음식물쓰레기에서 악취제거 효과가 뛰어났으며, 매우 빠른 속도로 악취를 제거하여, 악취 제거능이 우수한 균주이다. 동시에 본 발명의 균주는 또한 축산폐수 또는 음식물쓰레기내에 포함되어진 중금속의 일부에 대하여 매우 높은 수준으로 중금속을 제거하는 능력을 가지고 있다.The present invention relates to a new strain Bacillus megaterium BC2-1 KCCM 10858P useful for removing odors and heavy metals, heavy metals, odor removal, liquid fertilization method using food waste and a method of producing livestock feed using the same. will be. Bacillus megalium BC2-1 KCCM 10858P according to the present invention was excellent in removing odor from livestock wastewater and food waste, and is a strain having excellent odor removal ability by removing odor at a very high speed. At the same time, the strains of the present invention also have the ability to remove heavy metals at very high levels for some of the heavy metals contained in livestock wastewater or food waste.
Description
본 발명은 악취제거 및 중금속 제거 능력이 있는 바실러스 메가터리움 BC2-1 및 이를 이용한 음식물쓰레기 처리 방법에 관한 것이다.The present invention relates to Bacillus megaliumium BC2-1 having an ability to remove odors and heavy metals, and a method for treating food waste using the same.
가축을 집단 사육하는 경우 발생하는 축산분뇨가 축사의 피트내에서 저류하는 과정에 발생되는 악취 및 그로 인한 축사의 악취 등은 가축이나 사람에게 모두 유해한 물질이며 그 대부분이 지구온난화 가스로서 대기의 주요 오염물질이다. 이와 같은 악취는 주로 가축의 축분에서 기인되며 이들의 일반적인 조성을 보면 미생물에 의해서 쉽게 분해되는 탄수화물, 전분, 단백질 및 셀룰로오스 등의 이분해성 물질과 미생물에 의해서 분해가 잘 이루어지지 않는 지방, 리그닌 등의 난분해성 물질로 구분할 수 있다.Odors generated during livestock manure storage in the pit of the barn, and the resulting odors, which are harmful to both livestock and humans, are mostly global warming gases. It is a substance. These odors are mainly derived from livestock stocks, and their general composition shows that they are easily decomposed by microorganisms such as carbohydrates, starches, proteins and cellulose, and eggs that are not easily degraded by microorganisms such as fats and lignin. It can be divided into degradable substances.
또한, 축분이 혐호기 상태에서 분해되어 질 때 발생되는 악취성분은 암모니아, 메칠메캅탄, 황화수소, 황화메칠, 이황화메칠, 트리메칠아민, 아세트알데하이드, 스틸렌, 프로피온산, 낙산, 길초산 등이며 이들의 악취는 여러 가지 형태로 나타난다. 예를 들면, 메칠메캅탄은 썩은 양파 냄새, 황화수소는 무색의 기체로서 자극성이며, 질식성이면서 썩은 양파 냄새를 풍기며, 암모니아는 무색의 기체로서 자극성이며 뇨 냄새와 같은 특이취를 나타내는 등 이들 각각이 복합되면 복합취를 내어 사람의 위생상 혐오감을 주며 가축의 성장을 저해하게 된다.In addition, the odor components generated when the nutrient is decomposed in the aerobic state are ammonia, methylmecaptan, hydrogen sulfide, methyl sulfide, methyl disulfide, trimethylamine, acetaldehyde, styrene, propionic acid, butyric acid and gil acetic acid. Odors come in many forms. For example, methylmecaptan has a rotten onion odor, hydrogen sulfide is a colorless gas that is irritating, suffocating and rotting onion, and ammonia is a colorless gas that is irritating and has a characteristic odor like urine. If you do so, you will get a disgusting hygiene and inhibit livestock growth.
악취로 인한 불편은 현재 지속적으로 증가 추세에 있고, 법적 규제 또한 제정 및 시행되어 악취 관리의 중요성이 제기 되고 있다. 악취의 발생 원인으로는 축산폐수 처리장, 음식물쓰레기 처리장, 분뇨처리장, 매립장, 사료제조업 등으로 보고되고 있다. 그러나 이러한 시설에서 발생되는 악취는 일부공정에서만 발생되는 것이 아니라 전체 공정에서 발생되어, 악취발생 저감을 위한 공정관리, 시설개선, 방지시설 설치 등에 큰 어려움이 있다. 또한, 이런 환경기초시설 등은 주로 주거지역과 혼재하여 존재하므로 주변에 미치는 영향을 최소화하기 위해서는 효율이 높은 악취의 저감 기술 개발 등이 필요하다.Odor discomfort continues to increase at present, and legal regulations have been enacted and enforced to raise the importance of odor management. Odors are reported as livestock wastewater treatment plants, food waste treatment plants, manure treatment plants, landfills, and feed manufacturing industries. However, the odor generated in such a facility is not generated only in some processes but is generated in the entire process, and there is a great difficulty in process management, facility improvement, and prevention facility installation to reduce odor occurrence. In addition, since such environmental basic facilities are mainly mixed with residential areas, it is necessary to develop highly efficient odor reduction technology to minimize the impact on the surroundings.
현재 악취에 대한 법적 규제는 황화수소(H2S), 메틸메르캅탄(CH3SH), 황화메틸((CH3)2S), 이황화메틸((CH3)2S2), 암모니아(NH3), 트리메틸아민((CH3)3N), 아세트알데히드(CH3CHO), 스티렌(C6H5CH=CH2)등 8가지 항목에 대하여 시행되고 있으며, 이들 성분은 다른 유기화합물과 달리 저농도에서도 강한 냄새를 내는 것으로 알려져 있다. 특히, 돈사, 우사, 계사 등 축사에서 발생하는 악취는 황화수소(H2S)가 주된 오염물질이지만, 그 외에도 암모니아(NH3), 메틸메르캅탄(CH3SH), 트리메틸아민((CH3)3N) 및 저급지방산 등의 물질이 함유되어 악취의 강도를 상승시키게 된다.Legal regulations on odors currently include hydrogen sulfide (H 2 S), methyl mercaptan (CH 3 SH), methyl sulfide ((CH 3 ) 2 S), methyl disulfide ((CH 3 ) 2 S 2 ), and ammonia (NH 3 ), Trimethylamine ((CH 3 ) 3 N), acetaldehyde (CH 3 CHO), and styrene (C 6 H 5 CH = CH 2 ) are carried out on eight items. It is known to have a strong smell even at low concentrations. In particular, the odor generated in the barn such as pigs, barn, cages, etc., hydrogen sulfide (H 2 S) is the main pollutant, but in addition, ammonia (NH 3 ), methyl mercaptan (CH 3 SH), trimethylamine ((CH 3 ) 3 N) and contains a substance such as a lower fatty acid to thereby increase the strength of odor.
대개, 악취물질은 발생원별로 여러 가지 복합된 화합물이 원인으로 작용하여 악취를 유발하며, 단일화합물질로서 황화수소(H2S)는 마치 계란 썩는 냄새를 내고, 메르캅탄(Mercaptan)은 야채 썩는 냄새를 내며, 아민류는 생선냄새 등과 같은 특이한 냄새를 낸다.In general, malodorous substances cause various malodorous compounds by their source, causing malodors. As a single compound, hydrogen sulfide (H 2 S) smells like rotting eggs, and mercaptan smells like rotting vegetables. The amines have a peculiar smell like fish smell.
악취를 제거하는 방법으로는 크게 악취물질을 분리 또는 파괴시키는 방법으 로 산화법, 효소분해법 및 흡착법 등이 있고 단순히 악취를 은폐시키는 방법 인 소위 마스킹법이 있다. 산화법은 악취물질을 산화, 분해하고 박테리아, 곰팡이, 바이러스등에 대한 광범위한 살균 효과로 악취를 근원적으로 제거하는 방법으로써 그 동안 많은 방법이 제시되어왔다. 화학적인 산화법에서 많이 사용되어온 산화제의 종류로는 이산화염소(두오 존), 차아염소산 소다(락스) 및 이산화염소산염 등이 있으나 이러한 물질들은 염소를 함유한 물질로 인체 및 가축에 유해한 영향을 미칠 수 있다. 효소분해법은 식물엑기스를 추출하여 만든 탈취제로 냄새를 분해하는 작용이 있다고 하나 구체적인 성분 및 작용원리는 정확히 밝혀지지 않고 있으며 흡착법은 활성탄 등 표면적이 큰 흡착제를 이용하여 악취물질을 흡착시켜 제거하는 방법으로 일정 시간 사용하면 흡착제 표면적의 급속한 감소로 효율이 떨어지는 단점이 있다. 마스킹법은 천연 또는 인공향을 메틸알콜등 휘발성이 강한 용제에 녹인 후 대기 중에 휘산 시켜 악취를 은폐시켜 감각적인 악취문제는 해결할 수 있으나 악취의 근본적인 해결책이 되지는 못하고 있다.As a method of removing odors, there are largely methods of separating or destroying odorous substances, such as oxidation, enzymatic decomposition, and adsorption, and so-called masking, which simply hides odors. Oxidation has been suggested to oxidize and decompose odorous substances and to remove odors fundamentally with a wide range of bactericidal effects on bacteria, fungi and viruses. The types of oxidants that have been widely used in chemical oxidation methods include chlorine dioxide (duozone), sodium hypochlorite (Lax), and chlorine dioxide, but these substances contain chlorine and may have harmful effects on humans and livestock. . Enzymatic digestion is a deodorant made by extracting plant extracts, which has the effect of decomposing odors.However, specific components and principles of action are not known precisely.Adsorption is a method of adsorbing and removing odorous substances using an adsorbent having a large surface area such as activated carbon. When used for a certain time, the efficiency decreases due to the rapid decrease of the surface area of the adsorbent. The masking method solves the sensory odor problem by dissolving natural or artificial flavors in a volatile solvent such as methyl alcohol and then volatilizing it in the air, but it is not a fundamental solution to the odor.
화학물질을 사용하지 않는 방법으로 오존이나 자외선을 단독 혹은 병용하여 사용하는 방법이 있으나 오존의 경우 살균 및 탈취효과가 있지만 오존의 과다한 발생은 새로운 오염원으로 작용할 수 있다. 또한 오존 발생과 함께 TiO2와 같은 광촉매를 이용하여 악취원을 분해하는 방법이 제시 되었으나 TiO2와 같은 촉매는 비표면적을 늘리는데 한계가 있을 뿐 아니라 황화수소(H2S), 메틸메르캅탄(CH3SH), 황화메틸((CH3)2S), 이황화메틸((CH3)2S2)과 같은 황화물과 작용할 경우 촉매 표면이 쉽게 피독(Poisoning)되어 촉매 효율이 급격히 감소하는 단점을 가지고 있다.Although there is a method of using ozone or ultraviolet light alone or in combination as a method without using chemicals, ozone has a sterilizing and deodorizing effect, but excessive generation of ozone may act as a new pollutant. In addition, a method of decomposing odor sources using photocatalysts such as TiO 2 with ozone generation has been proposed, but catalysts such as TiO 2 have limitations in increasing the specific surface area, as well as hydrogen sulfide (H 2 S) and methyl mercaptan (CH 3). When interacting with sulfides such as SH), methyl sulfide ((CH 3 ) 2 S), and methyl disulfide ((CH 3 ) 2 S 2 ), the catalyst surface is easily poisoned and the catalyst efficiency decreases drastically. .
일반적으로 석유화학공장, 도시하수/분뇨처리장, 고무제조공장, 도색공장, 식품가공공장, 축산분뇨처리장, 분뇨처리장 등 각종 산업장에서 악취 및 벤젠(Benzene), 톨루엔(Toluene), 크실렌(Xylene) 등의 BTX류와 유기산, 알데히드류, 케톤류, 방향족화합물 등의 휘발성유기화합물(VOCs)이 동시에 대량으로 발생하는데, 이로 인한 주변 민원으로 산업발전에 필요한 공장의 건설이 지연되고, 도시의 기반시설이 되는 하수종말처리장 및 쓰레기매립장의 건설이 늦어지고 있다.In general, odor, benzene, toluene, and xylene in various industrial sites such as petrochemical plant, urban sewage / manure treatment plant, rubber manufacturing plant, painting plant, food processing plant, livestock manure treatment plant, and manure treatment plant. Volatile organic compounds (VOCs) such as BTX, organic acids, aldehydes, ketones, and aromatic compounds are generated in large quantities at the same time, which causes delays in the construction of factories necessary for industrial development due to surrounding complaints. Construction of sewage treatment plants and landfills is delayed.
산업현장에서 발생하는 악취 및 휘발성유기화합물(VOCs)을 제거하는 방법은 크게 화학적 방법과 생물학적 방법으로 나눌 수 있다. 화학적 방법으로 연소법은 2차 대기오염을 발생시키고, 흡착법은 고가인 활성탄을 사용함에 따라 비경제적이다. 이들 방법은 건설비가 적게 드는 대신 과다한 약품 소모로 운전비가 많이 소요된다. 이에 비하여 생물학적 방법은 건설비와 운전조건이 까다로운 단점을 가지고 있으나, 유지 및 관리가 용이하며 운전비용이 적게 소모된다. 생물학적 악취 제거 법은 담체에 악취 분해 미생물을 고정화시켜서 반응기에 충진한 바이오필터법 등이 있는데 바이오필터법은 경제적이고 2차 오염을 유발하지 않은 악취처리방법으로 부상하고 있다. 바이오필터시스템의 운전에 소용되는 운전비용은 가장 경제적이다. 운전에 소요되는 비용은 바이오필터, 약액세정, 촉매연소, 활성탄흡착, 직접연소의 순으로 많이 든다.Removal of odors and volatile organic compounds (VOCs) generated in industrial sites can be largely divided into chemical and biological methods. The chemical method of combustion produces secondary air pollution, and the adsorption method is uneconomical due to the use of expensive activated carbon. These methods cost a lot of money due to excessive chemical consumption instead of low construction costs. On the other hand, biological methods have disadvantages in that construction costs and operating conditions are difficult, but they are easy to maintain and manage and consume less operating costs. Biological odor removal methods include biofilters filled in reactors by immobilizing malodor-decomposing microorganisms on a carrier, and biofilters have emerged as an odor treatment method that is economical and does not cause secondary pollution. The operating costs for operating the biofilter system are the most economical. Operation costs are high in order of biofilter, chemical liquid cleaning, catalytic combustion, activated carbon adsorption, and direct combustion.
한국특허등록 10-1106030,10-1106032, 10-1106033, 10-1106034, 10-1106035, 및 10-1106037호에서는 바실러스 균주를 이용한 축산폐수 또는 음식물쓰레기의 자원화를 위한 정화방법"에 대한 특허가 등록되었다. Korean Patent Registration No. 10-1106030,10-1106032, 10-1106033, 10-1106034, 10-1106035, and 10-1106037 have registered a patent for "purification method for the recycling of livestock wastewater or food waste using Bacillus strain" It became.
본 발명은 가축사육시 발생되는 폐수 속의 악취 또는 음식물쓰레기에서 발생되는 악취 및 상기 폐수와 쓰레기속의 중금속을 감소시키기 위한 바실러스 메가터리움 BC2-1(Bacillus megaterium BC1-1) KCCM 10858P 균주를 제공하는 것을 목적으로 한다.The present invention provides a Bacillus megaterium BC1-1 KCCM 10858P strain for reducing the odor generated from wastewater or food waste generated in livestock raising and heavy metals in the wastewater and wastes. The purpose.
본 발명은 바실러스 메가터리움 BC2-1(Bacillus megaterium BC2-1) KCCM 10858P균주를 유효성분으로 함유하는 악취제거 및 중금속 제거용 미생물제제 및 이를 이용한 축산폐수 또는 음식물 쓰레기 자원화를 위한 정화 방법을 제공하는 것을 목적으로 한다.The present invention provides a microbial agent for removing odor and heavy metals containing Bacillus megaterium BC2-1 KCCM 10858P strain as an active ingredient and a method for purifying livestock wastewater or food waste using the same For the purpose of
본 발명은 종래의 균주들 보다 매우 빠르고 효율적으로 악취를 제거하는 능력이 있으며, 중금속 또한 제거하는 능력이 있는 바실러스 메가터리움 BC2-1(Bacillus megaterium BC2-1) KCCM 10858P 균주를 제공한다.The present invention provides a Bacillus megaterium BC2-1 KCCM 10858P strain that has the ability to remove odors much faster and more efficiently than conventional strains, and also has the ability to remove heavy metals.
본 발명은 바실러스 메가터리움 BC2-1(Bacillus megaterium BC2-1) KCCM 10858P균주를 유효성분으로 함유하는 악취제거 및 중금속 제거용 미생물제제를 제공한다.The present invention provides a microbial agent for odor removal and heavy metal removal containing Bacillus megaterium BC2-1 KCCM 10858P strain as an active ingredient.
본 발명에 따른 바실러스 메가터리움 BC2-1(Bacillus megaterium BC1-1) KCCM 10856P 균주는 축산 폐수 또는 음식물 쓰레기에서 발생되는 악취를 우수하게 감소시켰다. 특히 상기 미생물은 축산 폐수 또는 음식물 쓰레기에서 발생되는 중금속 또한 감소시켰다. 따라서 본 발명에 따른 미생물제제는 축산폐수 또는 음식물 쓰레기의 악취 및 중금속을 정화하는 데 있어서 사용될 수 있다. 또한 축산폐수를 이용한 유기질 액비 생산 또는 음식물 쓰레기를 이용한 가축사료 생산에 널리 사용될 수 있다.Bacillus megaterium BC1-1-1 according to the present invention KCCM 10856P strain excellently reduced the odor generated from livestock wastewater or food waste. In particular, the microorganisms also reduced heavy metals from livestock wastewater or food waste. Therefore, the microbial agent according to the present invention can be used to purify odors and heavy metals of livestock wastewater or food waste. In addition, it can be widely used for the production of organic liquid fertilizer using livestock wastewater or livestock feed using food waste.
본 발명에 따른 바실러스 메가터리움 BC2-1 KCCM 10858P는 축산폐수 및 음식물쓰레기에서 악취제거 효과가 뛰어났으며, 매우 빠른 속도로 악취를 제거하여, 악취 제거능이 우수한 균주이다. 동시에 본 발명의 균주는 또한 축산폐수 또는 음식물쓰레기내에 포함되어진 중금속의 일부에 대하여 매우 높은 수준으로 중금속을 제거하는 능력을 가지고 있다.Bacillus megalium BC2-1 KCCM 10858P according to the present invention was excellent in removing odor from livestock wastewater and food waste, and is a strain having excellent odor removal ability by removing odor at a very high speed. At the same time, the strains of the present invention also have the ability to remove heavy metals at very high levels for some of the heavy metals contained in livestock wastewater or food waste.
도 1은 축산 폐수의 악취 제거 특성 실험장치를 나타낸다.
도 2는 음식물 쓰레기의 악취 제거 특성 실험장치를 나타낸다. 1 shows an experimental apparatus for removing odor of livestock wastewater.
Figure 2 shows an experimental device for removing odor of food waste.
발명의 실시를 위한 최선의 형태Best Mode for Carrying Out the Invention
본 발명의 제 1 형태는 악취 제거 및 중금속 제거에 유용한 신균주 바실러스 메가테리움 BC2-1(Bacillus megaterium BC2-1) KCCM 10858P을 제공한다.A first aspect of the invention provides Bacillus megaterium BC2-1 KCCM 10858P, which is useful for odor removal and heavy metal removal.
본 발명의 제 2 형태는 신균주 바실러스 메가터리움 BC2-1 KCCM 10858P을 유효량 포함하고 있는 악취 제거 및 중금속제거용 미생물제제를 제공한다.A second aspect of the present invention provides a microbial agent for odor removal and heavy metal removal, containing an effective amount of the new strain Bacillus megaliumium BC2-1 KCCM 10858P.
보다 구체적으로, 본 발명은 바실러스 메가터리움 BC1-1 KCCM 10856P(Bacillus megaterium BC1-1), 바실러스 메가터리움 바실러스 메가터리움 BC1-2 KCCM 10857P(Bacillus megaterium BC1-2), 바실러스 세레우스 BC3 KCCM 10859P(Bacillus cereus BC3), 바실러스 라이케니포미스 BC4 KCCM 10860P(Bacillus licheniformis BC4), 바실러스 세레우스 BC5 KCCM 10861P(Bacillus cereus BC5), 바실러스 스파에리쿠스 BC6 KCCM 10862P(Bacillus sphaericus BC6), 바실러스 클라우시 BC7 KCCM 10863P(Bacillus clausii BC7), 바실러스 라이케니포미스 BC8 KCCM 10864P(Bacillus licheniformis BC8), 바실러스 퍼무스 BC9 KCCM 10865P(Bacillus firmus BC9), 바실러스 세레우스 BC10 KCCM 10866P(Bacillus cereus BC10)로 이루어진 그룹에서 선택되는 2 개 이상의 바실러스 종과 유효량의 신균주 바실러스 메가터리움 BC2-1 KCCM 10858P로 구성된 악취제거 및 중금속 제거용 미생물제제를 제공한다.More specifically, the present invention is Bacillus megaterium BC1-1 KCCM 10856P ( Bacillus megaterium BC1-1), Bacillus megaliumium Bacillus megaterium BC1-2 KCCM 10857P ( Bacillus megaterium BC1-2), Bacillus cereus BC3 KCCM 10859P (Bacillus cereus BC3), Bacillus rickeniformis BC4 KCCM 10860P ( Bacillus licheniformis BC4), Bacillus cereus BC5 KCCM 10861P ( Bacillus cereus BC5), Bacillus sphaericus BC6 KCCM 10862P ( Bacillus sphaericus BC6), Ba7 KCCM 10863P ( Bacillus clausii BC7), Bacillus rickeniformis BC8 KCCM 10864P ( Bacillus licheniformis BC8), Bacillus permus BC9 KCCM 10865P (Bacillus firmus BC9), Bacillus cereus BC10 KCCM 10866P (group selected from Bacillus cereus BC10) Determination of microorganisms for the removal of odors and heavy metals consisting of two or more Bacillus species and an effective amount of the new strain Bacillus megalium BC2-1 KCCM 10858P Ball.
본 발명의 제 3 의 형태는 신균주 바실러스 메가터리움 BC2-1 KCCM 10858P을 악취를 내는 음식물 쓰레기 또는 축산 폐수와 함께 배양하여 악취 및 음식물 쓰레기 또는 축산폐수 내의 중금속을 제거하는 방법을 제공한다. 보다 구체적으로 본 발명은 바실러스 메가터리움 BC1-1 KCCM 10856P(Bacillus megaterium BC1-1), 바실러스 메가터리움 바실러스 메가터리움 BC1-2 KCCM 10857P(Bacillus megaterium BC1-2), 바실러스 세레우스 BC3 KCCM 10859P(Bacillus cereus BC3), 바실러스 라이케니포미스 BC4 KCCM 10860P(Bacillus licheniformis BC4), 바실러스 세레우스 BC5 KCCM 10861P(Bacillus cereus BC5), 바실러스 스파에리쿠스 BC6 KCCM 10862P(Bacillus sphaericus BC6), 바실러스 클라우시 BC7 KCCM 10863P(Bacillus clausii BC7), 바실러스 라이케니포미스 BC8 KCCM 10864P(Bacillus licheniformis BC8), 바실러스 퍼무스 BC9 KCCM 10865P(Bacillus firmus BC9), 바실러스 세레우스 BC10 KCCM 10866P(Bacillus cereus BC10)로 이루어진 그룹에서 선택되는 2개 이상의 바실러스 종과 유효량의 신균주 바실러스 메가터리움 BC2-1 KCCM 10858P로 구성된 악취제거용 미생물제제를 악취는 내는 음식물 쓰레기 또는 축산폐수와 함께 배양하여 악취 및 음식물 쓰레기 또는 축산폐수 내의 중금속을 제거하는 방법을 제공한다.A third aspect of the present invention provides a method of removing the bad bacteria and food waste or heavy metals in livestock wastewater by culturing the new strain Bacillus megalium BC2-1 KCCM 10858P with malodorous food waste or livestock wastewater. More specifically, the present invention is Bacillus megaterium BC1-1 KCCM 10856P ( Bacillus megaterium BC1-1), Bacillus megaliumium Bacillus megacium BC1-2 KCCM 10857P ( Bacillus megaterium BC1-2), Bacillus cereus BC3 KCCM 10859P (Bacillus cereus BC3), Bacillus rickeniformis BC4 KCCM 10860P ( Bacillus licheniformis BC4), Bacillus cereus BC5 KCCM 10861P ( Bacillus cereus BC5), Bacillus sp. Erythropoid BC6 KCCM 10862P ( Bacillus sphaericus BC6) 10863P ( Bacillus clausii BC7), Bacillus rickeniformis BC8 KCCM 10864P ( Bacillus licheniformis BC8), Bacillus permus BC9 KCCM 10865P (Bacillus firmus BC9), Bacillus cereus BC10 KCCM 10866P ( Bacillus cereus BC10) Odor-depleting microbial agent composed of two or more Bacillus species and an effective amount of the new strain Bacillus megalium BC2-1 KCCM 10858P The present invention provides a method for removing heavy metals in odor and food waste or livestock waste by incubating with waste or livestock waste water.
본 발명의 제 4 의 형태는 신균주 바실러스 메가터리움 BC1-1 KCCM 10856P을 유효량 포함하고 있는 조성물을 이용한 축산 폐수를 이용한 유기질 액비 생산 방법을 제공한다. 보다 바람직하게는 본 발명은 바실러스 메가터리움 BC1-1 KCCM 10856P(Bacillus megaterium BC1-1), 바실러스 메가터리움 바실러스 메가터리움 BC1-2 KCCM 10857P(Bacillus megaterium BC1-2), 바실러스 세레우스 BC3 KCCM 10859P(Bacillus cereus BC3), 바실러스 라이케니포미스 BC4 KCCM 10860P(Bacillus licheniformis BC4), 바실러스 세레우스 BC5 KCCM 10861P(Bacillus cereus BC5), 바실러스 스파에리쿠스 BC6 KCCM 10862P(Bacillus sphaericus BC6), 바실러스 클라우시 BC7 KCCM 10863P(Bacillus clausii BC7), 바실러스 라이케니포미스 BC8 KCCM 10864P(Bacillus licheniformis BC8), 바실러스 퍼무스 BC9 KCCM 10865P(Bacillus firmus BC9), 바실러스 세레우스 BC10 KCCM 10866P(Bacillus cereus BC10)로 이루어진 그룹에서 선택되는 2개 이상의 바실러스 종과 유효량의 신균주 바실러스 메가터리움 BC2-1 KCCM 10858P로 구성된 조성물을 이용하여 축산 폐수를 이용한 유기질 액비 생산 방법을 제공한다.A fourth aspect of the present invention provides a method for producing organic liquid fertilizer using livestock wastewater using a composition containing an effective amount of new strain Bacillus megalium BC1-1 KCCM 10856P. More preferably, the present invention provides Bacillus megaterium BC1-1 KCCM 10856P, Bacillus megaterium BC1-1, Bacillus megaterium BC1-2 KCCM 10857P, Bacillus cereus BC3 KCCM 10859P (Bacillus cereus BC3), Bacillus rickeniformis BC4 KCCM 10860P ( Bacillus licheniformis BC4), Bacillus cereus BC5 KCCM 10861P ( Bacillus cereus BC5), Bacillus sphaericus BC6 KCCM 10862P ( Bacillus sphaericus BC6), Ba7 KCCM 10863P ( Bacillus clausii BC7), Bacillus rickeniformis BC8 KCCM 10864P ( Bacillus licheniformis BC8), Bacillus permus BC9 KCCM 10865P (Bacillus firmus BC9), Bacillus cereus BC10 KCCM 10866P (group selected from Bacillus cereus BC10) Using livestock wastewater with a composition consisting of at least two Bacillus species and an effective amount of the new strain Bacillus megalium BC2-1 KCCM 10858P. Provide quality manure production methods.
본 발명의 제 5 의 형태는 신균주 바실러스 메가터리움 BC2-1 KCCM 10858P을 유효량 포함하고 있는 조성물을 이용한 음식물 쓰레기를 이용한 가축사료 생산 방법을 제공한다. 보다 바람직하게는 본 발명은 바실러스 메가터리움 BC1-1 KCCM 10856P(Bacillus megaterium BC1-1), 바실러스 메가터리움 바실러스 메가터리움 BC1-2 KCCM 10857P(Bacillus megaterium BC1-2), 바실러스 세레우스 BC3 KCCM 10859P(Bacillus cereus BC3), 바실러스 라이케니포미스 BC4 KCCM 10860P(Bacillus licheniformis BC4), 바실러스 세레우스 BC5 KCCM 10861P(Bacillus cereus BC5), 바실러스 스파에리쿠스 BC6 KCCM 10862P(Bacillus sphaericus BC6), 바실러스 클라우시 BC7 KCCM 10863P(Bacillus clausii BC7), 바실러스 라이케니포미스 BC8 KCCM 10864P(Bacillus licheniformis BC8), 바실러스 퍼무스 BC9 KCCM 10865P(Bacillus firmus BC9), 바실러스 세레우스 BC10 KCCM 10866P(Bacillus cereus BC10)로 이루어진 그룹에서 선택되는 2 개 이상의 바실러스 종과 유효량의 신균주 바실러스 메가터리움 BC2-1 KCCM 10858P로 구성된 조성물을 이용한 음식물 쓰레기를 이용한 가축사료 생산 방법을 제공한다. A fifth aspect of the present invention provides a method for producing livestock feed using food waste using a composition containing an effective amount of the new strain Bacillus megalium BC2-1 KCCM 10858P. More preferably, the present invention provides Bacillus megaterium BC1-1 KCCM 10856P, Bacillus megaterium BC1-1, Bacillus megaterium BC1-2 KCCM 10857P, Bacillus cereus BC3 KCCM 10859P (Bacillus cereus BC3), Bacillus rickeniformis BC4 KCCM 10860P ( Bacillus licheniformis BC4), Bacillus cereus BC5 KCCM 10861P ( Bacillus cereus BC5), Bacillus sphaericus BC6 KCCM 10862P ( Bacillus sphaericus BC6), Ba7 KCCM 10863P ( Bacillus clausii BC7), Bacillus rickeniformis BC8 KCCM 10864P ( Bacillus licheniformis BC8), Bacillus permus BC9 KCCM 10865P (Bacillus firmus BC9), Bacillus cereus BC10 KCCM 10866P (group selected from Bacillus cereus BC10) Using food waste with a composition consisting of at least two Bacillus species and an effective amount of the new strain Bacillus megalium BC2-1 KCCM 10858P. Axis provides a feed production methods.
발명의 실시를 위한 형태Embodiment for Invention
이하 본 발명을 실시예를 통하여 상세하게 설명한다. Hereinafter, the present invention will be described in detail through examples.
<실시예 1> 바실러스 메가터리움 BC2-1(<Example 1> Bacillus megaliumium BC2-1 ( Bacillus megaterium Bacillus megaterium BC2-1) KCCM 10858P 균주의 분리 및 동정BC2-1) Isolation and Identification of KCCM 10858P Strains
본 발명에 사용되는 바실러스 메가터리움 BC2-1(Bacillus megaterium BC2-1) KCCM 10858P 균은 전라북도 익산 삼기면 숲에서 낙엽위에서 서식하는 버섯과 균사덩어리에서 얻어졌다. 채취되어진 버섯과 균사체를 500ml 비이커에 넣은 후 물 300ml와 흙설탕 약 5g 정도를 넣어서 제조된 배양액에 담구어 준 후 28 -30℃에서 항온 배양 하였다. 상기 배양액의 일부를 재례식 화장실에 뿌려준 후 하룻만에 악취가 사라지는 것을 보고, 본 발명의 바실러스 균이 악취제거능이 좋은 것을 알게되었다. 상기 배양액은 당밀을 사용하여서 계속적으로 배양되었으며, 이후에 음식물 쓰레기에 사용한 결과 악취 제거능이 우수한 것을 또한 확인하고, 유용 미생물의 순수분리에 들어갔다. Bacillus megaterium BC2-1 KCCM 10858P bacteria used in the present invention were obtained from mushrooms and mycelium masses that live on the leaves in the Samgimyeon forest, Iksan, Jeollabuk-do. The mushrooms and mycelium collected were put in a 500ml beaker, immersed in a culture solution prepared by adding 300ml of water and about 5g of earth sugar and incubated at 28 -30 ° C. After spraying a portion of the culture solution to the ceremonial toilet to see the odor disappears in one day, it was found that the Bacillus bacteria of the present invention has a good odor removal ability. The culture solution was continuously cultured using molasses, and subsequently used for food waste, it was also confirmed that the odor removal ability is excellent, and entered the pure separation of useful microorganisms.
유용미생물의 분리는 군산대학교 미생물학과에서 이루어졌으며, NA 배지를 이용하여서 총 12종의 미생물을 분리하였으며, 사슬형태의 배열을 가지고 내열성 포자를 형성하며, 페니실린에 내성을 보여서, 이들 모두가 바실러스 계통인 것을 확인하였다. 순수분리되어진 미생물은 NA배지를 이용하여서 계대하여 보관하였다. 순수 분리된 12종의 바실러스 균 모두의 16S rDNA 유전자 서열을 충남 대전의 한국생명공학원에 의뢰해 분석하였다. 그 결과 바실러스 메가터리움 BC2-1(Bacillus megaterium BC2-1) KCCM 10858P 종을 포함하여, 바실러스 메가터리움 BC1-1(Bacillus megaterium BC1-1) KCCM 10856P, 바실러스 메가터리움 BC1-2 KCCM 10857P(Bacillus megaterium BC1-2), 바실러스 세레우스 BC3 KCCM 10859P(Bacillus cereus BC3), 바실러스 라이케니포미스 BC4 KCCM 10860P(Bacillus licheniformis BC4), 바실러스 세레우스 BC5 KCCM 10861P(Bacillus cereus BC5), 바실러스 스파에리쿠스 BC6 KCCM 10862P(Bacillus sphaericus BC6), 바실러스 클라우시 BC7 KCCM 10863P(Bacillus clausii BC7), 바실러스 라이케니포미스 BC8 KCCM 10864P(Bacillus licheniformis BC8), 바실러스 퍼무스 BC9 KCCM 10865P(Bacillus firmus BC9), 바실러스 세레우스 BC10 KCCM 10866P(Bacillus cereus BC10) 종으로 동정되었다. 이를 2007년 4월 27일에 한국종균센터에 기탁하고, 각각의 KCCM 기탁번호를 부여받았다.Separation of useful microorganisms was performed at the Department of Microbiology, Kunsan National University, and a total of 12 microorganisms were isolated using NA medium, forming a heat resistant spore with a chain-like arrangement, and exhibiting resistance to penicillin, all of which are Bacillus strains. It confirmed that it was. The pure microorganisms were passaged and stored using NA medium. The 16S rDNA gene sequences of all 12 isolated Bacillus bacteria were analyzed by the Korea Institute of Biotechnology of Daejeon, Chungnam. As a result, including Bacillus megaterium BC2-1 KCCM 10858P species, Bacillus megaterium BC1-1 KCCM 10856P, Bacillus megaterium BC1-2 KCCM 10857P ( Bacillus megaterium BC1-2), Bacillus cereus BC3 KCCM 10859P ( Bacillus cereus BC3), Bacillus rickeniformis BC4 KCCM 10860P ( Bacillus licheniformis BC4), Bacillus cereus BC5 KCCM 10861P ( Bacillus cereus BC5), Bacillus sp. KCCM 10862P (Bacillus sphaericus BC6), Bacillus claw when BC7 KCCM 10863P (Bacillus clausii BC7) , Bacillus Lai Kenny Po Ms BC8 KCCM 10864P (Bacillus licheniformis BC8) , Bacillus buffer mousse BC9 KCCM 10865P (Bacillus firmus BC9) , Bacillus cereus BC10 KCCM 10866P ( Bacillus cereus BC10) species was identified. It was deposited with the Korean spawn center on April 27, 2007 and each KCCM accession number was assigned.
<실시예 2> 바실러스 메가터리움 BC2-1 KCCM 10858P 및 바실러스 메가터리움 BC2-1 KCCM 10858P와 다른 바실러스 균의 혼합미생물 조성물에 의한 축산폐수의 악취제거 효과Example 2 Deodorization Effect of Livestock Wastewater by Mixed Microbial Composition of Bacillus Megacium BC2-1 KCCM 10858P and Bacillus Megacium BC2-1 KCCM 10858P
바실러스 메가터리움 BC2-1 KCCM 10858P에 의한 축산 폐수의 악취 저감 효율을 측정하기 위하여 바실러스 메가터리움 BC2-1 KCCM 10858P과 바실러스 메가터리움 BC1-1(Bacillus megaterium BC1-1) KCCM 10856P, 바실러스 메가터리움 BC1-2 KCCM 10857P(Bacillus megaterium BC1-2), 바실러스 세레우스 BC3 KCCM 10859P(Bacillus cereus BC3), 바실러스 라이케니포미스 BC4 KCCM 10860P(Bacillus licheniformis BC4), 바실러스 세레우스 BC5 KCCM 10861P(Bacillus cereus BC5), 바실러스 스파에리쿠스 BC6 KCCM 10862P(Bacillus sphaericus BC6), 바실러스 클라우시 BC7 KCCM 10863P(Bacillus clausii BC7), 바실러스 라이케니포미스 BC8 KCCM 10864P(Bacillus licheniformis BC8), 바실러스 퍼무스 BC9 KCCM 10865P(Bacillus firmus BC9), 바실러스 세레우스 BC10 KCCM 10866P(Bacillus cereus BC10) 혼합 미생물군을 일정한 축산 폐수가 들어있는 미생물 반응용기(도 1 참조)에 투입하여 4일 경과 후에 용기 내의 악취를 포집하여 측정하였다. Bacillus mega emitter Solarium BC2-1 KCCM 10858P by Bacillus in order to measure the odor reduction efficiency of livestock waste Mega emitter Solarium BC2-1 KCCM 10858P and Bacillus mega emitter Solarium BC1-1 (Bacillus megaterium BC1-1) KCCM 10856P , Bacillus mega Bacillus certer BC3-2, Bacillus cereus BC3 KCCM 10859P ( Bacillus cereus BC3), Bacillus rickeniformis BC4 KCCM 10860P ( Bacillus licheniformis BC4), Bacillus cereus BC5 KCCM 10861P ( Bacillus cereus BC5), Bacillus spa Erie kusu BC6 KCCM 10862P (Bacillus sphaericus BC6) , Bacillus claw when BC7 KCCM 10863P (Bacillus clausii BC7) , Bacillus Lai Kenny Po Ms BC8 KCCM 10864P (Bacillus licheniformis BC8) , Bacillus buffer mousse BC9 KCCM 10865P (Bacillus firmus BC9), Bacillus cereus BC10 KCCM 10866P ( Bacillus cereus BC10) mixed microbial population is placed in a microbial reaction vessel (see Fig. 1 ) containing a constant livestock wastewater After 4 days, the odor in the container was collected and measured.
축산폐수의 미생물 반응용기 조건은 다음과 같이 미생물의 활발한 활동을 위하여 포기기를 설치하여 일정한 유량의 포기와 가열밴드(heating band)와 자동온도 조절 장치를 이용하여 온도를 37.5℃로 일정하게 유지시켜 주었다(도 1 참조). The microbial reaction vessel condition of livestock wastewater was set up to maintain the temperature at 37.5 ℃ by using aeration and heating band and thermostat to install aeration device for active microorganism as follows. (See FIG. 1 ).
또한, 미생물 투입 전에 축산폐수 원수에서 발생되는 악취농도를 측정하여 미생물 투입 후 악취 항목별 농도 변화 추이를 분석하였다. 정확한 효율 측정을 위하여 축산 폐수의 악취 저감을 위한 11개의 미생물 반응 용기에 각각 2리터씩 폐수를 주입한 후, 1개의 용기에는 바실러스 메가터리움 BC2-1 KCCM 10858P 배양액 5ml, 또다른 용기에는 바실러스 메가터리움 BC1-1 KCCM 10856P(Bacillus megaterium BC1-1), 바실러스 메가터리움 BC1-2 KCCM 10857P(Bacillus megaterium BC1-2), 바실러스 메가터리움 BC2-1 KCCM 10858P(Bacillus megaterium BC2-1), 바실러스 세레우스 BC3 KCCM 10859P(Bacillus cereus BC3), 바실러스 라이케니포미스 BC4 KCCM 10860P(Bacillus licheniformis BC4), 바실러스 세레우스 BC5 KCCM 10861P(Bacillus cereus BC5), 바실러스 스파에리쿠스 BC6 KCCM 10862P(Bacillus sphaericus BC6), 바실러스 클라우시 BC7 KCCM 10863P(Bacillus clausii BC7), 바실러스 라이케니포미스 BC8 KCCM 10864P(Bacillus licheniformis BC8), 바실러스 퍼무스 BC9 KCCM 10865P(Bacillus firmus BC9), 바실러스 세레우스 BC10 KCCM 10866P(Bacillus cereus BC10) 배양액이 균등하게 혼합된 혼합미생물 5ml투입, 나머지 1개의 용기에는 미생물을 투입하지 않고 폐수 5ml을 넣어서 대조 폐수로 사용하였다. In addition, the concentration of odor generated in the livestock wastewater before microbial input was measured and analyzed for the change in concentration of each odor after microbial input. For accurate measurement of efficiency, 2 liters of wastewater were injected into each of 11 microbial reaction vessels to reduce the odor of livestock wastewater, and one vessel of 5 ml of Bacillus megalium BC2-1 KCCM 10858P culture medium and another vessel of Bacillus mega Bacillus megaterium BC1-1, Bacillus megaterium BC1-2, Bacillus megaterium BC1-2, Bacillus megaterium BC1-2, Bacillus megaterium BC1-2 Cereus BC3 KCCM 10859P ( Bacillus cereus BC3), Bacillus rickeniformis BC4 KCCM 10860P ( Bacillus licheniformis BC4), Bacillus cereus BC5 KCCM 10861P ( Bacillus cereus BC5), Bacillus sphericus BC6 KCCM 10862P ( Bacillus spha) Bacillus claw when BC7 KCCM 10863P (Bacillus clausii BC7) , Bacillus Lai Kenny Po Ms BC8 KCCM 10864P (Bacillus licheniformis BC8) , Bacillus buffer mousse BC9 KCCM 10865P (Bacillus firmus BC9) , Bacillus Aureus KCCM 10866P BC10 (Bacillus cereus BC10) culture media were used as evenly mixed mixture microorganism 5ml turned on, the remaining one container without added microorganisms contrast put the waste water effluent 5ml.
(1) 분석 방법(1) analysis method
1) 암모니아의 측정법1) Measurement method of ammonia
분석용 시료용액에 페놀-니트로프루시드 나트륨용액과 차아염소산 나트륨용액을 가하고 암모늄이온과 반응시켜 생성되는 인도 페놀류의 흡광도를 측정하여 암모니아를 정량한다.Ammonia is quantified by measuring the absorbance of indophenols produced by adding phenol-nitroprusside sodium solution and sodium hypochlorite solution to the analytical sample solution and reacting with ammonium ion.
가. 시약end. reagent
- 채취용액-Collecting solution
붕산 5 g을 증류수에 녹여 전량을 1 L로 한다.5 g of boric acid is dissolved in distilled water, and the total amount is 1 L.
- 페놀, 펜타시아노 니트로실 철(Ⅲ)산 나트륨 용액-Phenol, pentacyano nitrosyl sodium (III) acid solution
페놀 5 g, 펜타시아노 니트로실 암모늄 철(Ⅲ) 나트륨 2수화물6) 25 mg 을 증류수에 용해하여 전량을 500 mL로 한다. 이 용액은 차고 어두운 곳에 보존하고 조제한 후 1 개월 이상 경과한 것은 사용하지 않는다.5 g of phenol and 25 mg of pentacyano nitrosyl ammonium iron (III) sodium dihydrate6) are dissolved in distilled water to make a total amount of 500 mL. Store this solution in a cool dark place and do not use more than 1 month after preparation.
- 차아염소산 나트륨 용액Sodium hypochlorite solution
차아염소산 나트륨 용액(유효염소 3∼10 %) 60/CmL[여기에서 C는 조제시에 정량한 차아염소산 나트륨(NaOCl)의 유효염소 농도(단위 %)]와 수산화나트륨 10 g및 인산수소 나트륨 12 수화물 35.8 g을 증류수에 용해하여 전량을 1 L로 한다. 이 용액은 사용할 때 마다 제조한다.Sodium hypochlorite solution (3-10% effective chlorine) 60 / CmL (where C is the effective chlorine concentration (in%) of sodium hypochlorite (NaOCl) quantified at the time of preparation)], 10 g sodium hydroxide and sodium hydrogen phosphate 12 Dissolve 35.8 g of hydrate in distilled water to make the total amount 1 L. This solution is prepared each time it is used.
나. 용액흡수I. Solution absorption
흡수병은 흡수액 용량 20 mL를 담을 수 있는 경질유리제로 여과구가 장치되어 있는 것을 사용한다. 중간에 흡수액 용량 20 mL를 담을 수 있는 채취용액을 넣어 2개를 직렬로 연결시키고, 흡인펌프는 흡수 병을 장치한 상태에서 10 L/분 이상의 공기를 흡인 한다. 채취 후 2 개의 흡수병중의 흡수용액을 합하여 용량 50 mL 메스플라스크에 옮기고,다시 흡수병의 내부를 포집용액으로 씻은 용액을 메스플라스크에 옮기고 전량을 50 mL로 한다. 이 용액 10 mL 를 시험관에 옮기고 분석용 시료용액으로 사용한다.The absorber bottle is made of hard glass that can hold 20 mL of absorbent liquid and is equipped with a filter port. In the middle, take a sample solution that can hold 20 mL of absorbent liquid and connect the two in series. The suction pump draws more than 10 L / min of air with an absorbent bottle installed. After collection, the absorbent solutions from the two absorbent bottles are combined and transferred to a 50 mL volumetric flask, and the solution washed with the collection solution is transferred to the volumetric flask and the total amount is 50 mL. Transfer 10 mL of this solution into the test tube and use it as the sample solution for analysis.
다. 분석용 시료용액의 흡광도 측정All. Absorbance Measurement of Analytical Sample Solution
분석용 시료용액에 페놀 펜타시아노 니트로실 철(Ⅲ)산 나트륨 용액 5 mL를 가하여 잘 흔들어 섞은 후, 차아염소산나트륨 용액 5 mL를 혼합하여 25∼30 ℃에서 1 시간 방치한 후, 640 nm 파장에서 흡광도를 측정한다. 공 시험액은 포집용액(흡수액) 10 mL 를 분석용 시료용액과 같은 방법으로 조작하여 사용한다.Add 5 mL of phenol pentacyano nitrosyl iron (III) solution to the sample for analysis, shake well, and mix 5 mL of sodium hypochlorite solution for 1 hour at 25 to 30 ° C. Absorbance is measured at. For blank test solution, use 10 mL of the collecting solution (absorbent solution) in the same manner as the analytical sample solution.
2) 메틸머캅탄, 황화수소, 다이메틸설파이드 및 다이메틸다이설파이드 시험방법 2) Methylmercaptan, hydrogen sulfide, dimethyl sulfide and dimethyl disulfide test method
황화물 분석법은 시료가스를 -170℃ SPT(Sample Preconcentration Trap)로 저온 흡착농축 시켜, 이것을 다시 가열하여 탈착시켜 가스크로마토그래프에 주입시키며 검출기로는 PFPD(Pulse Flame Photometri Dectector)를 사용하였다. In the sulfide method, the sample gas was concentrated at -170 ° C. in a sample preconcentration trap (SPT) at low temperature, heated again, desorbed, and injected into a gas chromatograph, and a PFPD (Pulse Flame Photometri Dectector) was used as a detector.
3) 트리메틸아민3) trimethylamine
트리메틸아민의 분석 방법은 황산수용액(359+1)에 흡수시킨 분석용 시료 용액을 정량 주사기로 분취하여 수산화칼륨 용액이 들어 있는 분해병 안에 실리콘 마개를 통하여 주입한 후 0.2-0.L/min 의 질소 2-3L 정도 버블링하여 발생된 트리메틸아민을 액체 산로로 냉각시켜 농축관에 농축시킨 후 70℃ 정도까지 가열 탈착시켜 트리메틸아민을 컬럼으로 도입하여 FID로 분석하였다. Trimethylamine was analyzed by diluting the sample solution for analysis absorbed in aqueous sulfuric acid solution (359 + 1) with a syringe, and injecting it through a silicone stopper into a decomposition bottle containing potassium hydroxide solution. The trimethylamine generated by bubbling about 2-3 L of nitrogen was cooled with a liquid acid, concentrated in a condenser tube, heated and desorbed to about 70 ° C., and trimethylamine was introduced into a column and analyzed by FID.
4) 알데하이드의 분석4) Analysis of Aldehydes
알데하이드류의 분석방법은 2,4-디니트로페닐하이드라진(2.4-dinitrophenyl hydrazine)을 피복시킨 옥타데실 실릴화 실리카겔을 충진시킨 시료포집관에 통과시켜 알데하이드류를 포집하여 시료 포집관으로부터 아세토니트릴을 증발시킨 후 아세트산에틸에 녹여 그 일부를 HPLC(영린)에 도입하여 정량하였다. Analysis of aldehydes is carried out through a sample collection tube filled with octadecyl silylated silica gel coated with 2,4-dinitrophenyl hydrazine, to collect aldehydes and to evaporate acetonitrile from the sample collection tube. The mixture was dissolved in ethyl acetate, and a portion thereof was introduced into HPLC (Young Lin) for quantification.
(2) 분석결과(2) Analysis result
바실러스 메가터리움 BC1-1 KCCM 10856P 및 바실러스 메가터리움 BC1-1 KCCM 10856P와 다른 바실러스 균의 혼합미생물 조성물을 넣고 4일 배양한 것과 대조폐수만을 넣은 것에 대한 결과를 하기 표 1 (단위 ppm)에 나타내었다. Bacillus megagium BC1-1 KCCM 10856P and Bacillus megagium BC1-1 KCCM 10856P and the mixed microbial composition of the other Bacillus bacteria incubated for 4 days and the control waste water only in Table 1 (unit ppm) Indicated.
바실러스 메가터리움 BC2-1 KCCM 10858P은 축산폐수에서 암모니아, 황화수소, 아세트알데히드를 높게 제거하였다. 다만, 메틸멀캅탄, 다이메틸다이설파이드는 투입 전의 농도와 투입 4일 후 대조시료, 혼합미생물 투입 시료가 거의 유사한 농도로 검출되었다. 그러나 다이메틸설파이드는 대조시료 대비 소량증가하였다. Bacillus megacium BC2-1 KCCM 10858P removes ammonia, hydrogen sulfide and acetaldehyde from livestock wastewater. However, methyl mercaptan and dimethyl disulfide were detected at almost the same concentration as the concentration before the addition, the control sample and the mixed microorganism after 4 days. Dimethyl sulfide, however, increased slightly in comparison to the control sample.
<실시예 3> 바실러스 메가터리움 BC1-1 KCCM 10856P 및 바실러스 메가터리움 BC1-1 KCCM 10856P과 다른 바실러스 균의 혼합미생물 조성물에 의한 음식물쓰레기의 악취 분석 결과Example 3 Odor Analysis of Food Waste by Mixed Microorganism Composition of Bacillus Megacium BC1-1 KCCM 10856P and Bacillus Megacium BC1-1 KCCM 10856P
바실러스 메가터리움 BC2-1 KCCM 10858P에 의한 음식물 쓰레기의 악취 저감 효율을 측정하기 위하여 바실러스 메가터리움 BC2-1 KCCM 10858P과 바실러스 메가터리움 BC1-1(Bacillus megaterium BC1-1) KCCM 10856P, 바실러스 메가터리움 BC1-2 KCCM 10857P(Bacillus megaterium BC1-2), 바실러스 세레우스 BC3 KCCM 10859P(Bacillus cereus BC3), 바실러스 라이케니포미스 BC4 KCCM 10860P(Bacillus licheniformis BC4), 바실러스 세레우스 BC5 KCCM 10861P(Bacillus cereus BC5), 바실러스 스파에리쿠스 BC6 KCCM 10862P(Bacillus sphaericus BC6), 바실러스 클라우시 BC7 KCCM 10863P(Bacillus clausii BC7), 바실러스 라이케니포미스 BC8 KCCM 10864P(Bacillus licheniformis BC8), 바실러스 퍼무스 BC9 KCCM 10865P(Bacillus firmus BC9), 바실러스 세레우스 BC10 KCCM 10866P(Bacillus cereus BC10) 혼합 미생물군을 일정한 음식물 쓰레기가 들어있는 미생물 반응용기(도 2 참조)에 투입하여 4일 경과 후에 용기 내의 악취를 포집하여 측정하였다. Bacillus megaterium BC2-1 KCCM 10858P and Bacillus megaterium BC1-1 KCCM 10856P, Bacillus mega to measure the odor reduction efficiency of food waste by Bacillus megacrum BC2-1 KCCM 10858P Bacillus certer BC3-2, Bacillus cereus BC3 KCCM 10859P ( Bacillus cereus BC3), Bacillus rickeniformis BC4 KCCM 10860P ( Bacillus licheniformis BC4), Bacillus cereus BC5 KCCM 10861P ( Bacillus cereus BC5), Bacillus spa Erie kusu BC6 KCCM 10862P (Bacillus sphaericus BC6) , Bacillus claw when BC7 KCCM 10863P (Bacillus clausii BC7) , Bacillus Lai Kenny Po Ms BC8 KCCM 10864P (Bacillus licheniformis BC8) , Bacillus buffer mousse BC9 KCCM 10865P (Bacillus firmus BC9), Bacillus cereus BC10 KCCM 10866P ( Bacillus cereus BC10) mixed microbial population in a microbial reaction vessel containing a certain food waste (see Figure 2 ) After 4 days of ingestion, the odor in the container was collected and measured.
음식물 쓰레기의 미생물 반응용기 조건은 점성이 강하여 가열밴드(heating band)와 자동온도 조절 장치를 이용하여 온도를 37.5℃로 일정하게 유지시켜 주었다(도 2 참조). Microbial reaction vessel conditions of food waste was viscous to maintain a constant temperature to 37.5 ℃ using a heating band (heating band) and a thermostat (see Figure 2 ).
또한, 미생물 투입 전에 생음식쓰레기에서 발생되는 악취농도를 측정하여 미생물 투입 후 악취 항목별 농도 변화 추이를 분석하였다. 정확한 효율 측정을 위하여 음식물 쓰레기의 악취 저감을 위한 11개의 미생물 반응 용기에 각각 2리터씩 일정량 주입한 후 1개의 용기에는 바실러스 메가터리움 BC2-1 KCCM10858P 배양액 5ml, 또다른 용기에는 바실러스 메가터리움 BC1-1 KCCM10856(Bacillus megaterium BC1-1), 바실러스 메가터리움 BC1-2 KCCM 10857P(Bacillus megaterium BC1-2), 바실러스 메가터리움 BC2-1 KCCM 10858P(Bacillus megaterium BC2-1), 바실러스 세레우스 BC3 KCCM 10859P(Bacillus cereus BC3), 바실러스 라이케니포미스 BC4 KCCM 10860P(Bacillus licheniformis BC4), 바실러스 세레우스 BC5 KCCM 10861P(Bacillus cereus BC5), 바실러스 스파에리쿠스 BC6 KCCM 10862P(Bacillus sphaericus BC6), 바실러스 클라우시 BC7 KCCM 10863P(Bacillus clausii BC7), 바실러스 라이케니포미스 BC8 KCCM 10864P(Bacillus licheniformis BC8), 바실러스 퍼무스 BC9 KCCM 10865P(Bacillus firmus BC9), 바실러스 세레우스 BC10 KCCM 10866P(Bacillus cereus BC10) 배양액이 균등하게 혼합된 혼합미생물 5ml투입, 나머지 1개의 용기에는 미생물을 투입하지 않고 음식물 쓰레기 5ml을 넣어서 대조군으로 사용하였다.In addition, the concentration of odor generated by raw food waste was measured before microbial input, and the concentration change of each odor after microbial input was analyzed. In order to accurately measure the odor of food waste, two liters of 11 microbial reaction vessels were injected into each of the microbial reaction vessels, and one container of Bacillus megalium BC2-1 KCCM10858P 5 ml of culture medium and another container of Bacillus megalium BC1 -1 KCCM10856 ( Bacillus megaterium BC1-1), Bacillus megaterium BC1-2 KCCM 10857P ( Bacillus megaterium BC1-2), Bacillus megaliumium BC2-1 KCCM 10858P ( Bacillus megaterium BC2-1), Bacillus cereus BC3 KCCM 10859P ( Bacillus cereus BC3), Bacillus rickeniformis BC4 KCCM 10860P ( Bacillus licheniformis BC4), Bacillus cereus BC5 KCCM 10861P ( Bacillus cereus BC5), Bacillus sphaericus BC6 KCCM 10862P ( Bacillus sphaericus BC6), Ba7 KCCM 10863P ( Bacillus clausii BC7), Bacillus rickeniformis BC8 KCCM 10864P ( Bacillus licheniformis BC8), Bacillus permus BC9 KCCM 10865P ( Bacillus firmus BC9), Bacillus Serum BC10 KCCM 10866P ( Bacillus cereus BC10) 5 ml of mixed microorganisms in which the culture solution was evenly mixed, and the other one container was used as a control without putting microorganisms 5ml of food waste.
음식물 쓰레기의 악취 분석은 실시예 2의 축산 폐수 악취 분석 실험방법과 동일한 방법으로 수행하였다.Odor analysis of food waste was carried out in the same manner as the livestock wastewater odor analysis experimental method of Example 2.
<결과><Result>
바실러스 메가터리움 BC2-1 KCCM10858은 음식물쓰레기에서 암모니아, 황화수소, 메틸멀캅탄, 다이메틸설파이드는 투입 전의 농도와 투입 4일 후 대조시료, 혼합미생물 투입 시료가 거의 유사한 농도로 검출되어 다이메틸설파이드, 아세트 알데히드에 대한 제거 효율은 미미한 것으로 보인다. 다이메틸다이설파이드은 투입 전의 농도와 비교하였을 때 소량 감소하였고, 대조시료 대비는 큰 폭으로 감소하였다.Bacillus megacium BC2-1 KCCM10858 has a similar concentration of ammonia, hydrogen sulfide, methyl mercaptan, and dimethyl sulfide in food waste, and the control sample and mixed microbial input sample after 4 days of injection. Removal efficiency for acetaldehyde appears to be negligible. Dimethyldisulfide decreased in a small amount compared to the concentration before the input, and significantly decreased compared to the control sample.
<실시예 4> 축산폐수 및 음식물쓰레기의 중금속 저감효과 분석Example 4 Analysis of Heavy Metal Reduction Effect of Livestock Wastewater and Food Waste
바실러스 메가터리움 BC2-1 KCCM10858P에 의한 축산폐수 및 음식물 쓰레기에서의 중금속 저감 효과를 측정하기 위하여 11개의 미생물 반응 용기에 음식물쓰레기 각각 2리터씩 일정량 주입한 후 1개의 용기에는 바실러스 메가터리움 BC2-1 KCCM10858P 배양액 5ml, 나머지 1개의 용기에는 미생물을 투입하지 않고 음식물 쓰레기 5ml을 넣어서 대조군으로 사용하였다. 축산폐수에 대하여서도 같은 방법으로 실시하였다. 미생물 배양시 및 배양 4일후에 음식물쓰레기 및 축산폐수에서 중금속의 농도를 측정하였다. 중금속의 측정방법은 다음과 같은 방법으로 수행하였다.Bacillus Megalium BC2-1 In order to measure the effect of reducing heavy metals in livestock wastewater and food waste by KCCM10858P, 11 liters of food waste were injected into each of 11 microbial reaction vessels, and then one container of Bacillus megalium BC2- 1 KCCM10858P culture solution 5ml, the other one container was used as a control without putting microorganisms 5ml of food waste. The same was done for livestock wastewater. The concentration of heavy metals in food waste and livestock wastewater was measured during microbial culture and after 4 days of culture. Heavy metal was measured by the following method.
(1) 시약의 제조(1) Preparation of Reagent
- 터너리 용액(Ternary solution): HNO3, H2SO4, HClO4를 각각 10:1:4 비율로 혼합하여 필요한 양 만큼 준비한다.Ternary solution: Mix HNO 3 , H 2 SO 4 and HClO 4 in the ratio of 10: 1: 4 to prepare the required amount.
- 농축 HNO3 -Concentrated HNO 3
(2) 분해(2) disassembly
채취한 시료 10ml을 250ml 삼각플라스크에 평량한 후 농축 HNO3 10-30ml을 가한 후 하룻밤 방치한 다음 핫플레이트(Hot plate)에 올려 놓고 서서히 가열하여 흰 침전이 생길 때까지 건조시킨다. 냉각한 후 터너리 용액 10-50ml을 가하여 핫플레이트에서 가열하여 H2SO4 및 HClO4의 흰 연기를 어느 정도 날려 보낸 후 분해액이 백색이 되거나 갈색으로 투명하게 되면 분해를 멈춘다. 냉각 시킨 다음 뜨거운 증류수를 가해 50 또는 100ml 매스플라스크에 No. 6 여과지를 사용하여 여과하고 계속 뜨거운 증류수로 분해 플라스크를 폴리스멘(policemen)을 사용하여 깨끗이 씻어 여과한다.10 ml of the collected sample is weighed into a 250 ml Erlenmeyer flask, 10-30 ml of concentrated HNO 3 is added, and left to stand overnight, then placed on a hot plate and dried slowly until a white precipitate is formed. After cooling, 10-50 ml of ternary solution is added and heated on a hot plate to blow off some white smoke of H 2 SO 4 and HClO 4 , and the decomposition stops when the decomposition solution becomes white or brown. After cooling, add hot distilled water to the No. 50 or 100 ml mass flask. 6 Filter with filter paper and continue with hot distilled water. Rinse the flask with policemen and filter.
(3) 측정(3) measurement
여액은 일정량을 취하여 희석한 후 K, Ca, Mg, Na, Fe, Mn, Cu, Zn, Cr, Cd, Pb, Ni 등을 AA기(Atomic Absorption Flame Emission Spectrometer) 또는 ICP(Inductively Coupled Plasma)에서 각 성분의 표준용액으로 검량선을 작성한 후 측정한다.The filtrate is diluted in a predetermined amount, and then K, Ca, Mg, Na, Fe, Mn, Cu, Zn, Cr, Cd, Pb, Ni, and the like are collected in AA (Atomic Absorption Flame Emission Spectrometer) or ICP (Inductively Coupled Plasma). Make a calibration curve with the standard solution of each component and measure it.
<결과><Result>
(1) 아연의 제거효과(1) Removal effect of zinc
배양후 4일째 초기 369 ppm에서 4일째에는 17.2ppm으로서 95.4%의 아연 제거효율을 보였다(표 3 참조)After 4 days of culture, the zinc removal efficiency of 95.4% was obtained from the initial 369 ppm at 4 days to 17.2 ppm (see Table 3).
(2) 구리의 제거효과(2) copper removal effect
배양후 4일째 초기 86.3 ppm에서 4일째에는 2.6ppm으로서 97%의 구리 제거효율을 보였다(표 4 참조)At 46.3 days after incubation, it was 2.6ppm at the initial 86.3 ppm and showed 97% copper removal efficiency (see Table 4).
(3) 납의 제거효과(3) lead removal effect
배양후 4일째 초기 0.73ppm에서 4일째에는 0.03ppm으로서 95.9%의 납 제거효율을 보였다(표 5참조)The lead removal efficiency of 95.9% was seen from 0.73ppm at 4 days after culture to 0.03ppm at 4 days (see Table 5).
따라서 본 발명에 따른 Bacillus megaterium BC2-1은 중금속의 제거에 효과적이었다.Therefore, Bacillus megaterium BC2-1 according to the present invention was effective in removing heavy metals.
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| KR102551904B1 (en) * | 2022-12-26 | 2023-07-06 | (주)에코비즈넷 | Bacillus licheniformis BS and composition for reducing oder containing the same |
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