KR101106035B1 - Bacillus firmus bc9 10865p reducing nasty odor and heavy metal and clarification methods for resource recovery from food waste or livestock waste water by using it - Google Patents
Bacillus firmus bc9 10865p reducing nasty odor and heavy metal and clarification methods for resource recovery from food waste or livestock waste water by using it Download PDFInfo
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- KR101106035B1 KR101106035B1 KR1020097025248A KR20097025248A KR101106035B1 KR 101106035 B1 KR101106035 B1 KR 101106035B1 KR 1020097025248 A KR1020097025248 A KR 1020097025248A KR 20097025248 A KR20097025248 A KR 20097025248A KR 101106035 B1 KR101106035 B1 KR 101106035B1
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- bacillus
- cereus
- megaterium
- food waste
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Abstract
본 발명은 신균주 바실러스 퍼무스 BC9 KCCM 10865P를 포함하는 악취제거 및 중금속 제거 조성물 및 이를 이용한 축산폐수 또는 음식물쓰레기 정화방법에 관한 것으로서, 더욱 상세하게는 신균주 바실러스 퍼무스 BC9 KCCM 10865P 또는 이들을 유효성분으로 함유하는 미생물제제 및 이를 이용한 축산폐수 또는 음식물 쓰레기 자원화를 위한 정화방법에 관한 것이다.The present invention relates to a odor removal and heavy metal removal composition comprising the new strain Bacillus permus BC9 KCCM 10865P and a method for purifying livestock wastewater or food waste using the same, and more particularly, the new strain Bacillus permus BC9 KCCM 10865P or an active ingredient thereof. It relates to a microbial agent containing and a method for purifying livestock wastewater or food waste using the same.
Description
본 발명은 축산폐수 및 음식물쓰레기의 악취 및 중금속를 저감시키는 미생물을 제공하는 것을 목적으로 한다.An object of the present invention is to provide a microorganism which reduces odor and heavy metals of livestock wastewater and food waste.
또한 축산폐수를 이용한 유기질 액비 생산방법 및 음식물쓰레기를 이용한 가축사료 생산방법을 제공하는 것을 목적으로 한다.It is also an object of the present invention to provide a method for producing organic liquid fertilizer using livestock wastewater and a livestock feed production method using food waste.
본 발명은 가축사육시 발생되는 폐수 속의 악취 또는 음식물쓰레기에서 발생되는 악취 및 상기 폐수와 쓰레기 속의 중금속을 감소시키기 위한 미생물 및 이를 이용한 축산폐수 또는 음식물 쓰레기 자원화를 위한 정화 방법에 관한 것이다.The present invention relates to a microorganism for reducing odor generated from wastewater or food waste generated in livestock raising and heavy metals in the wastewater and garbage, and to a method for purifying livestock wastewater or 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 is a rotten onion smell, hydrogen sulfide is a colorless gas, 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), 아세트 알데히드(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), styrene (C 6 H 5 CH = CH 2 ) is carried out for eight items, these components are different from other organic compounds 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 방법으로 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 흡착 법은 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, but it has a function of decomposing odors.However, specific components and principles of action are not known precisely.Adsorption is a method of adsorbing and removing malodorous substances by using an adsorbent having a large surface area such as activated carbon. When used for a certain time, there is a disadvantage that 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.
한국특허출원번호 10-1996-0025606호(1996년 6월 29일 출원)에서는 음식물찌꺼기의 생물학적 처리용 발효제 조성물 및 퇴비화 방법에 관한 것으로서, 고온에 적응시킨 내고온성 사상균과; 바실러스 균에 속하는 내고온성 및 고온성 단백질 및 유지 분해균과; 고온성 방성균과; 고온에 적응시킨 내고온성 방선균을 포함하는 음식물 찌꺼기의 고속 발효 처리용 발효제 조성물을 제공하고 있다.Korean Patent Application No. 10-1996-0025606 (filed June 29, 1996) relates to a fermentation agent composition and a composting method for biological treatment of food wastes, high temperature resistant filamentous fungi; High temperature resistant and high temperature proteins belonging to Bacillus bacteria and oil-fat decomposition bacteria; Thermophilic fungi; Provided is a fermentation agent composition for fast fermentation treatment of food waste containing high temperature resistant actinomycetes adapted to high temperature.
한국등록특허 292879호(1999년 4월 7일 출원)는 축분처리용 미생물 제제 및 이를 이용한 축분의 처리방법에 관한 것으로서, 축분의 탄수화물과 단백질 처리효소 분비능이 탁월한 간상균(바실러스 서브틸리스(Bacillus subtilis KCTC 1028)과 그 분해물을 영양원으로 하는 악취를 제거하는 능력을 갖는 광합성 미생물(한국과학기술원 유전자 은행에 KCTC 8937P로 기탁된 광합성 미생물인 로도슈도모나스속 (Rhodopseudomonas sp.))로 이루어진 미생물 제제와 상기 미생물 제제를 축분과 축사에 적용하므로서 악취를 제거하고, 결과적으로 가축의 생육환경과 축산가의 생활환경을 개선하는 기능을 갖는 축분의 처리방법을 제공하고 있다.Korean Patent No. 292879 (filed April 7, 1999) relates to a microbial preparation for condensation treatment and a method of treating condensate using the same, and has an excellent ability to secrete carbohydrates and protein processing enzymes of condensate (Bacillus subtilis ( Bacillus subtilis) subtilis KCTC 1028) and photosynthetic microorganisms (Rhodopseudomonas sp., a photosynthetic microorganism deposited as KCTC 8937P with the Korea Advanced Institute of Science and Technology Gene Bank) By applying microbial preparations to livestock and livestock farms, there is provided a method for treating livestock, which has the function of removing odors and consequently improving the livestock environment and livestock environment of livestock.
한국등록특허 378667호(2000년 8월 31일 출원)는 가축분뇨 등의 신속한 무취화 및 퇴비화 방법 및 그 조성물에 관한 것으로서, 가축분뇨에서 발생하는 악취나 음식물 쓰레기등의 부패초기에 발생되는 부패취를 제거하고, 신속히 발효 분해시켜 무취화된 유기질 비료를 만드는데 사용되는 조성물 및 그 제조방법에 관한 것이다. 스트렙토미세스 속(Streptomyces sp.), 바실러스 속(Bacillus sp.), 마이크로콕커스 속(Micrococcus sp.), 아크로모박터 속(Achromobacter sp.), 후라보박테리움 속(Flavobacterium sp.), 에쎄리씨아 속(Escherichia sp.) 및 슈도모나스 속(Pseudomonas sp.)에서 선택된 1종 이상의 균주를 뉴트리엔트배지나 TSB배지와 같은 일반적인 액체배지에서 호기적으로 배양하여 석회석 분말, 벤트나이트 점토 또는 부식질 토양에서 선택된 흡착매체에 흡착시킨후 건조시켜서 얻어지는 무취화 조성물을 제공하고 있다.Korean Patent No. 378667 (filed Aug. 31, 2000) relates to a method for rapid deodorization and composting and composition of livestock manure, and is a decay odor generated in the early stages of decay such as odor or food waste generated from livestock manure. The present invention relates to a composition and a method for manufacturing the same, which are used to remove the organic matter and rapidly fermentatively decompose to form an odorless organic fertilizer. Streptomyces sp., Bacillus sp., Micrococcus sp., Achromobacter sp., Flavobacterium sp. One or more strains selected from the genus Escherichia sp. And Pseudomonas sp. Are aerobicly cultured in common liquid media such as neutrient or TSB media to select them from limestone powder, bentite clay or humus soil. An odorless composition obtained by adsorbing on an adsorption medium and then drying is provided.
한국등록특허 433268호(2000년 10월 5일 출원)는 악취물질의 생물학적 처리용 발효제 조성물 및 그 제조방법에 관한 것으로서, 고농도 유기물과 고온에 적응시킨 내고온성 악취유용 사상균과; 기질천이관계를 고려하고 악취유용성이 매우 우수한 사상균과; 바실러스 균에 속하는 내고온성 및 고온성이며 광범위 기질이용성을 가진 기질 분해균과 악취 유용균과; 고농도 유기물에 적응성을 가지면서 고온에 적응시킨 내고온성 방선균을 포함하는 악취유용 미생물들로 이루어진 발효제 조성 물 및 그 제조방법에 관한 것이다. 상기 내고온성 악취유용 사상균은, 아스퍼질러스 니거(Aspergillus niger), 아스퍼질러스 오리재(Aspergillus oryzae), 페니실리움 에스피피(Penicillium spp.), 뮤코르 에스피피(Mucor spp.), 트리쵸데르마 비리드(Trichoderma viride), 체팔로스포리움 아크레모니움(Cephallosporium acremonium), 아이싸첸키아 오리엔탈리스(Issatchenkia orientalis)로 이루어진 군에서 선택되는 1종 이상의 혼합균이 바람직하며, 상기 기질천이 관계를 고려하고 악취유용성이 매우 우수한 사상균은, 캔디다 에스피피(Candida spp.), 아르쓰로박터 에스피피(Arthrobacter spp.), 노카르디아 에스피피(Nocardia spp.)로 이루어진 군에서 선택되는 1종 이상의 혼합균을 이용하였고, 상기 바실러스 균에 속하는 내고온성 및 고온성이며 광범위 기질이용성을 가진 기질분해균과 악취 유용균은, 바실러스 서브틸리스 시에이취-10(Bacillus subtilis CH-10), 바실러스 메센테리커스(Bacillus mesentericus), 바실러스 메가터리움(Bacillus megaterium), 바실러스 리체니포르미스 엔엘알아이-33(Bacillus licheniformmis NLRI-33), 바실러스 투링지엔시스 (Bacillus thuringiensis), 바실러스 서큐란스(Bacillus circulans)로 이루어진 군에서 선택되는 1종 이상의 혼합균이 이용되었으며, 상기 고농도 유기물에 적응성을 가진 고온성 방선균은, 써모액티노마이세스 에스피(Thermoactinomyces sp.) 및/또는 써모몬스포라 에스피피(Thermomonspora spp.)인 발효제 조성물을 제공하였다. 그리고, 상기 고농도 유기물에 적응성을 가지면서 고온에 적응시킨 내고온성 방선균은, 스트렙토마이세스 그리슈스(Streptomyces griseus), 스트렙토마이세스 글로비스포리어스(Streptomyces globisporius), 스트렙토마이세스 플라부스 (Streptomyces flavus)로 이루어진 군으로부터 선택되는 1종 이상의 혼합균인 것이 바람직하며, 상기 광범위 기질이용성을 가진 기질 분해균과 악취 유용균은, 프슈도모나스 푸티다 엔엘알아이-엑스50(Pseudomonas putida NLRI-X50), 코리네박테리움 에스피피(Corynebacterium spp.), 스타필로코쿠스 렌투스(Staphylococcus lentus), 마이크로코쿠스 리래(Micrococcus lylae)로 이루어진 군에서 선택되는 1종 이상의 혼합균인 것이 바람직하다고 밝히고 있다.Korean Patent No. 433268 (filed Oct. 5, 2000) relates to a fermentation agent composition for biological treatment of malodorous substances and a method for producing the same, and includes a high temperature organic substance and a high temperature resistant malodorous filamentous fungus; Filamentous fungi with consideration of substrate transition and excellent odor availability; Substrate degrading bacteria and odor useful bacteria belonging to Bacillus bacteria having high temperature and high temperature and broad substrate availability; It relates to a fermentation agent composition consisting of microorganisms for odor oil containing high temperature resistant actinomycetes adapted to high temperature while being adapted to high concentration organic matter and a method of manufacturing the same. The high temperature odor-resistant filamentous fungus is Aspergillus niger , Aspergillus oryzae , Penicillium spp., Mucor spp., Tricho At least one mixed bacterium selected from the group consisting of Trichoderma viride , Cephallosporium acremonium , and Issatchenkia orientalis is preferred, and the substrate transition relationship The filamentous fungi that are highly contemplated and odor-soluble are at least one selected from the group consisting of Candida spp., Arthrobacter spp., And Nocardia spp. Mixed bacteria were used, and the high temperature resistant, high temperature resistant, broad-substrate usable substrate degrading bacteria and odor useful bacteria belong to Bacillus subtilis. Odor -10 (Bacillus subtilis CH-10) , Bacillus mesen Terry carcass (Bacillus mesentericus), Bacillus mega emitter Solarium (Bacillus megaterium), Bacillus piece you miss formate enel alahyi -33 (Bacillus licheniformmis NLRI-33) , Bacillus turing if N-Sys ( Bacillus thuringiensis ), one or more mixed bacteria selected from the group consisting of Bacillus circulans was used, the thermophilic actinomycetes adaptable to the high concentration organic matter, Thermoactinomyces sp. And / or thermomonspora spp. In addition, the high temperature resistant actinomycetes adapted to the high temperature while being adaptable to the high concentration organic material are Streptomyces griseus , Streptomyces globisporius , Streptomyces flavus It is preferable that it is at least one mixed bacterium selected from the group consisting of, and the above-mentioned substrate degrading bacteria and odor useful bacteria having broad substrate availability are Pseudomonas putida NLRI-X50 and Corynebacterium. Corynebacterium spp., Staphylococcus lentus ( Staphylococcus lentus ), Micrococcus lylae ( Micrococcus lylae ) is one or more selected from the group consisting of mixed bacteria is said to be preferable.
한국특허출원번호 10-2005-0044120호(2005년 5월 25일 출원)는 축사내 악취 및 황사 제거용 공기 정화장치에 관한 것으로 바실러스 서브틸러스를 이용하여 악취제거에 응용한 바 있다.Korean Patent Application No. 10-2005-0044120 (filed May 25, 2005) relates to an air purifier for removing odors and yellow dust in barns and has been applied to remove odors using a Bacillus subtilis.
한국특허출원번호 10-2006-0032543(2006년 4월 11일 출원)호에서는 광촉매반응기, 유동상 호기 및 혐기조 및 바이오필터로 조합된 하이브리드 시스템 공정을 이용하여 악취 폐가스를 효율적으로 처리하기 위한 방법 및 장치가 출원되었다.Korean Patent Application No. 10-2006-0032543 (filed April 11, 2006) discloses a method for efficiently treating malodorous waste gas using a hybrid system process including a photocatalytic reactor, a fluidized bed aerobic system, and an anaerobic tank and a biofilter. The device has been filed.
한국특허출원 10-2004-0011349호(출원일 2004년 2월 2일)에서는 지오트리쿰 속 균주가 고정된 바이오필터 및 이를 이용한 메틸에틸케톤의 제거방법을 제공하고 있다. 담체에 지오트리쿰 속 균주가 고정화된 바이오필터를 사용하여 메틸에틸케톤을 제거하는 경우, 활성탄이나 소각 등의 전통적인 방법에 비하여 상대적으로 저렴하고, 질소산화물이나 2차 오염물질을 생성하지 않았다고 하였다.Korean Patent Application No. 10-2004-0011349 (filed February 2, 2004) provides a biofilter to which the strain of the genus Geotricum is fixed and a method of removing methyl ethyl ketone using the same. When the methyl ethyl ketone was removed using a biofilter immobilized with a strain of Geotricum on the carrier, it was relatively inexpensive and did not generate nitrogen oxides or secondary pollutants compared to conventional methods such as activated carbon or incineration.
한국특허출원 10-2003-0078402호(출원일 2003년 11월 6일)에서는 악취 및 휘발성유기화합물을 제거를 위한 바이오필터를 제공하고 있으며, 슈도모나스(Pseudomonas)속의 균주를 이용하여 휘발성 유기화합물을 제거하는 것을 보인 바 있다.Korean Patent Application No. 10-2003-0078402 (filed Nov. 6, 2003) provides a biofilter for removing odors and volatile organic compounds, and removes volatile organic compounds using a strain of Pseudomonas genus. It has been shown.
기술적 과제Technical Challenge
본 발명은 가축사육시 발생되는 폐수 속의 악취 또는 음식물쓰레기에서 발생되는 악취 및 상기 폐수와 쓰레기 속의 중금속을 감소시키기 위한 바실러스 퍼무스 BC9(Bacillus firmus BC9) KCCM 10865P 균주를 제공하는 것을 목적으로 한다.It is an object of the present invention to provide a Bacillus firmus BC9 KCCM 10865P strain for reducing odor generated from wastewater or food waste generated in livestock raising and heavy metals in the wastewater and waste.
본 발명은 바실러스 퍼무스 BC9(Bacillus firmus BC9) KCCM 10865P 균주를 유효성분으로 함유하는 악취제거 및 중금속 제거용 미생물제제 및 이를 이용한 축산폐수 또는 음식물 쓰레기 자원화를 위한 정화 방법을 제공하는 것을 목적으로 한다.An object of the present invention is to provide a microbial agent for removing odor and heavy metals containing Bacillus firmus BC9 KCCM 10865P strain as an active ingredient and a method for purifying livestock wastewater or food waste using the same.
기술적 해결방법Technical solution
본 발명은 종래의 균주들 보다 매우 빠르고 효율적으로 악취를 제거하는 능력이 있으며, 중금속 또한 제거하는 능력이 있는 바실러스 퍼무스 BC9(Bacillus firmus BC9) KCCM 10865P 균주를 제공한다.The present invention provides a Bacillus firmus BC9 KCCM 10865P 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 퍼무스 BC9(Bacillus firmus BC9) KCCM 10865P 균주를 유효성분으로 함유하는 악취제거 및 중금속 제거용 미생물제제를 제공한다.The present invention provides a microbial agent for odor removal and heavy metal removal containing Bacillus firmus BC9 KCCM 10865P strain as an active ingredient.
본 발명에 따른 바실러스 퍼무스 BC9(Bacillus firmus BC9) KCCM 10865P 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 널리 사용될 수 있다. 또한 축산폐수를 이용한 유기질 액비 생산 또는 음식물 쓰레기를 이용한 가축사료 생산에 널리 사용될 수 있다. Bacillus firmus BC9 (Cacillus firmus BC9) KCCM 10865P strain according to the present invention excellently reduced the odor generated in 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 widely used for purifying 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.
도 1은 축산 폐수의 악취 제거 특성 실험장치를 나타낸다. 1 shows an experimental apparatus for removing odor of livestock wastewater.
도 2는 음식물 쓰레기의 악취 제거 특성 실험장치를 나타낸다. Figure 2 shows an experimental device for removing odor of food waste.
발명의 실시를 위한 최선의 형태Best Mode for Carrying Out the Invention
본 발명은 악취 제거 및 중금속 제거에 유용한 신균주 바실러스 퍼무스 BC9(Bacillus firmus BC9) KCCM 10865P 균주를 제공한다.The present invention provides a strain of Bacillus firmus BC9 KCCM 10865P, which is useful for removing odors and removing heavy metals.
본 발명은 또 다른 양태는 신균주 바실러스 퍼무스 BC9 KCCM 10865P를 유효량 포함하고 있는 악취 제거 및 중금속제거용 미생물제제를 제공한다.Another aspect of the present invention provides a microbial agent for odor removal and heavy metal removal comprising an effective amount of the new strain Bacillus permus BC9 KCCM 10865P.
본 발명에 따른 바실러스 퍼무스 BC9 KCCM 10865P를 제제화하는 방법은 종래의 기술을 응용할 수 있다. 미생물을 제제화하는 방법은 비드(bead) 형태의 고체로 제조하는 방법과, 액상제제로 제조하는 방법이 있다. 고체형태로 미생물제제를 제조하는 경우, 제조하는 과정에서 제품이 불균일하게 될 수 있으며, 미생물의 활성저하로 인한 제품의 적용성 감소 또는 제조공정상에 분진이 발생하는 등의 단점이 있다. 한편, 액상의 미생물제제는 분말제품에 비해 활성이 빠르고 취급이 간편한 등의 장점이 있지만, 분말제품에 비해 보존성이 떨어지는 단점이 있다.The method for formulating Bacillus permus BC9 KCCM 10865P according to the present invention can apply conventional techniques. Methods for formulating microorganisms include a method of preparing a solid in the form of beads and a method of preparing a liquid formulation. In the case of preparing the microorganism in a solid form, the product may be non-uniform in the manufacturing process, there is a disadvantage such as reduced application of the product due to the deactivation of microorganisms or dust generated in the manufacturing process. On the other hand, the liquid microbial agent has advantages such as faster activity and easier handling than the powder product, but has a disadvantage of poor shelf life compared to the powder product.
비드 형태로 제조하는 방법의 일례로는 (a) 알지네이트(alginate), 펙틴(pectin), 카라지난(carrageenan) 또는 폴리아스파르트산(polyaspartic acid)과 같 은 음전하 생분해성 고분자의 0.1∼5.0 중량% 용액과 바실러스 세레우스 BC3 KCCM 10859P 배양액의 혼합물을 Ca2+, Cu2+, Zn2+, Ni2+, Co2+, Mn2+, Al2+, Fe2+, 또는 Mg2+을 포함하는 0.5∼3.0 중량% 농도의 금속염 용액에 떨어뜨려 비드(bead)를 제조하는 단계; 및An example of a method of preparing in the form of beads is (a) 0.1-5.0 wt% solution of negatively charged biodegradable polymers such as alginate, pectin, carrageenan or polyaspartic acid. And a mixture of Bacillus cereus BC3 KCCM 10859P culture solution containing Ca 2+ , Cu 2+ , Zn 2+ , Ni 2+ , Co 2+ , Mn 2+ , Al 2+ , Fe 2+ , or Mg 2+ . Preparing a bead by dropping it in a metal salt solution at a concentration of 0.5-3.0 wt%; And
(b) 상기 비드를 키토산, 키토산 유도체 또는 폴리라이신(polylysine)과 같은 양전해 생분해성 고분자의 0.1∼3.0 중량% 용액에 침지시켜 30분∼2시간동안 교반시켜 비드를 정전기적 인력에 의해 코팅하는 단계를 포함하여 제조될 수 있다. 상기와 같이 제조된 미생물 제제를 동결건조하여 그대로 사용할 수도 있고, 미생물의 생존 및 활성효과의 지속성을 높이기 위하여; (c) 상기 비드를 50mM 소듐 아세테이트(pH 5.5)로 세척하여 여분의 양전하 생분해성 고분자 물질을 제거하는 단계; (d) 상기 비드를 0.1 내지 2.0 중량% 음전하 생분해성 고분자 용액에 침지시켜 교반하는 단계; 및 (e) 상기 비드를 생리식염수로 세척한 다음 0.1 내지 2.0 중량% 양전하 생분해성 고분자 용액에 침지시켜 교반하여 외막을 형성하는 단계를 추가로 실시하여 그 외부에 생분해성 고분자 막을 좀더 두텁게 형성하는 것에 의해 제조되는 되는 미생물 제제의 효과 지속성을 증대시킬 수 있다.(b) the beads are immersed in a 0.1 to 3.0% by weight solution of a positively-charged biodegradable polymer such as chitosan, chitosan derivatives or polylysine and stirred for 30 minutes to 2 hours to coat the beads by electrostatic attraction. It can be prepared including the step. The microbial preparation prepared as described above may be used as it is by lyophilization, in order to increase the persistence of the survival and active effect of the microorganism; (c) washing the beads with 50 mM sodium acetate (pH 5.5) to remove excess positively charged biodegradable polymeric material; (d) immersing the beads in 0.1-2.0 wt% negative charge biodegradable polymer solution and stirring; And (e) further washing the beads with physiological saline and then immersing in 0.1 to 2.0 wt% positive charge biodegradable polymer solution to stir to form an outer membrane to form a thicker biodegradable polymer membrane thereon. It is possible to increase the effect persistence of the microbial agent produced by.
본 발명의 또 다른 양태는 바실러스 메가터리움 BC1-1 KCCM 10856P(Bacillus megaterium BC1-1 KCCM 10856P), 바실러스 메가터리움 BC1-2 KCCM 10857P(Bacillus megaterium BC1-2 KCCM 10857P), 바실러스 메가터리움 BC2-1 KCCM 10858P(Bacillus megaterium BC2-1 KCCM 10858P), 바실러스 세레우스 BC3 KCCM 10859P(Bacillus cereus BC3 KCCM 10859P), 바실러스 라이케니포미스 BC4 KCCM 10860P(Bacillus licheniformis BC4 KCCM 10860P), 바실러스 세레우스 BC5 KCCM 10861P(Bacillus cereus BC5 KCCM 10861P), 바실러스 스파에리쿠스 BC6 KCCM 10862P(Bacillus sphaericus BC6 KCCM 10862P), 바실러스 클라우시 BC7 KCCM 10863P(Bacillus clausii BC7 KCCM 10863P), 바실러스 라이케니포미스 BC8 KCCM 10864P(Bacillus licheniformis BC8 KCCM 10864P), 바실러스 세레우스 BC10 KCCM 10866P(Bacillus cereus BC10 KCCM 10866P)으로 이루어진 그룹에서 선택되는 하나 이상의 바실러스 종과 유효량의 신균주 바실러스 퍼무스 BC9 KCCM 10865P로 구성된 악취제거 및 중금속 제거용 미생물제제를 제공하는 것이다.Another embodiment of the present invention is Bacillus megaterium BC1-1 KCCM 10856P ( Bacillus megaterium BC1-1 KCCM 10856P), Bacillus megaterium BC1-2 KCCM 10857P ( Bacillus megaterium BC1-2 KCCM 10857P), Bacillus megaliumium BC2 -1 KCCM 10858P (Bacillus megaterium BC2-1 KCCM 10858P), Bacillus cereus BC3 KCCM 10859P (Bacillus cereus BC3 KCCM 10859P), Bacillus Lai Kenny Po Ms BC4 KCCM 10860P (Bacillus licheniformis BC4 KCCM 10860P), Bacillus cereus BC5 KCCM 10861P (Bacillus cereus BC5 KCCM 10861P), Bacillus spa Erie kusu BC6 KCCM 10862P (Bacillus sphaericus BC6 KCCM 10862P), Bacillus claw when BC7 KCCM 10863P (Bacillus clausii BC7 KCCM 10863P), Bacillus Lai Kenny Po Ms BC8 KCCM 10864P (Bacillus licheniformis BC8 KCCM 10864P), Bacillus cereus BC10 KCCM 10866P) and at least one Bacillus species selected from the group consisting of Bacillus cereus BC10 KCCM 10866P It provides a microbial agent for odor removal and heavy metal removal composed of Permus BC9 KCCM 10865P.
본 발명의 또 다른 양태는 바실러스 퍼무스 BC9 KCCM 10865P를 악취를 내는 음식물 쓰레기 또는 축산 폐수와 함께 배양하여 악취 및 음식물 쓰레기 또는 축산폐수 내의 중금속을 제거하는 방법이다.Another aspect of the invention is a method of culturing Bacillus permus BC9 KCCM 10865P with malodorous food waste or livestock wastewater to remove odors and heavy metals in food waste or livestock wastewater.
본 발명의 또다른 양태는 바실러스 메가터리움 BC1-1 KCCM 10856P(Bacillus megaterium BC1-1 KCCM 10856P), 바실러스 메가터리움 BC1-2 KCCM 10857P(Bacillus megaterium BC1-2 KCCM 10857P), 바실러스 메가터리움 BC2-1 KCCM 10858P(Bacillus megaterium BC2-1 KCCM 10858P), 바실러스 세레우스 BC3 KCCM 10859P(Bacillus cereus BC3 KCCM 10859P), 바실러스 라이케니포미스 BC4 KCCM 10860P(Bacillus licheniformis BC4 KCCM 10860P), 바실러스 세레우스 BC5 KCCM 10861P(Bacillus cereus BC5 KCCM 10861P), 바실러스 스파에리쿠스 BC6 KCCM 10862P(Bacillus sphaericus BC6 KCCM 10862P), 바실러스 클라우시 BC7 KCCM 10863P(Bacillus clausii BC7 KCCM 10863P), 바실러스 라이케니포미스 BC8 KCCM 10864P(Bacillus licheniformis BC8 KCCM 10864P), 바실러스 세레우스 BC10 KCCM 10866P(Bacillus cereus BC10 KCCM 10866P)으로 이루어진 그룹에서 선택되는 하나 이상의 바실러스 종과 유효량의 신균주 바실러스 퍼무스 BC9 KCCM 10865P로 구성된 악취제거용 미생물제제를 악취는 내는 음식물 쓰레기 또는 축산폐수와 함께 배양하여 악취 및 음식물 쓰레기 또는 축산폐수 내의 중금속을 제거하는 방법이다.Another embodiment of the present invention Bacillus megaterium BC1-1 KCCM 10856P ( Bacillus megaterium BC1-1 KCCM 10856P), Bacillus megaterium BC1-2 KCCM 10857P ( Bacillus megaterium BC1-2 KCCM 10857P), Bacillus megaliumium BC2 -1 KCCM 10858P (Bacillus megaterium BC2-1 KCCM 10858P), Bacillus cereus BC3 KCCM 10859P (Bacillus cereus BC3 KCCM 10859P), Bacillus Lai Kenny Po Ms BC4 KCCM 10860P (Bacillus licheniformis BC4 KCCM 10860P), Bacillus cereus BC5 KCCM 10861P (Bacillus cereus BC5 KCCM 10861P), Bacillus spa Erie kusu BC6 KCCM 10862P (Bacillus sphaericus BC6 KCCM 10862P), Bacillus claw when BC7 KCCM 10863P (Bacillus clausii BC7 KCCM 10863P), Bacillus Lai Kenny Po Ms BC8 KCCM 10864P (Bacillus licheniformis BC8 KCCM 10864P), Bacillus cereus BC10 KCCM 10866P (Bacillus cereus BC10 KCCM 10866P) one or more of the novel strain Bacillus species and an effective amount selected from the group consisting of Bacillus Mousse the malodor removing microbial preparations consisting BC9 KCCM 10865P odor is a method for culturing with that food waste, or livestock waste water to remove heavy metals in the odor and food waste, or livestock waste water.
본 발명의 또 다른 양태는 신균주 바실러스 퍼무스 BC9 KCCM 10865P를 유효량 포함하고 있는 축산 폐수를 이용한 유기질 액비 생산 방법 및 음식물 쓰레기를 이용한 가축사료 생산 방법이다.Another embodiment of the present invention is an organic liquid fertilizer production method using livestock wastewater containing an effective amount of the new strain Bacillus permus BC9 KCCM 10865P and a livestock feed production method using food waste.
본 발명에 따른 축산폐수를 이용한 유기질 액비 생산 방법 및 음식물 쓰레기를 이용한 가축사료 생산 방법은 신균주 바실러스 퍼무스 BC9 KCCM 10865P를 사용하는 것 이외에 종래의 기술을 응용할 수 있다. 축산 폐수를 이용한 유기질 액비료 생산의 예를 들면, 축산폐수와 토탄(peat)을 이용한 유기질액비료의 제조방법이 있다. 이 방법은 토탄 및 유기물질과 굴껍질(oyster) 및 연탄재를 분쇄하여 적정비율로 혼합하는 단계; 탄산수소나트륨, 산화칼슘, 염화칼슘을 적정비율 혼합하여 탈취하는 단계; 미생물을 이용하여 발효시키는 단계; 유기질액비료를 추출하는 단계; 상기 추출된 유기질 액비료 에 묽은 염산을 첨가하여 중화시키는 단계; 및 상기 중화된 유기질액을 일정시간 폭기시켜 화학적 산소요구량(C.O.D)를 감소시키는 단계로 이루어져 있다.Organic liquid fertilizer production method using livestock wastewater and livestock feed production method using food waste according to the present invention can be applied to the conventional technology in addition to using the new strain Bacillus permus BC9 KCCM 10865P. Examples of organic liquid fertilizer production using livestock wastewater include a method for producing organic liquid fertilizer using livestock wastewater and peat. This method comprises the steps of pulverizing peat and organic material and oyster shells and briquettes at an appropriate ratio; Deodorizing by mixing a proper ratio of sodium bicarbonate, calcium oxide and calcium chloride; Fermentation using microorganisms; Extracting the organic liquid fertilizer; Neutralizing by adding dilute hydrochloric acid to the extracted organic liquid fertilizer; And aeration of the neutralized organic liquid for a predetermined time to reduce the chemical oxygen demand (C.O.D).
음식물 쓰레기를 이용한 발효사료화는 신균주 바실러스 퍼무스 BC9 KCCM 10865P를 이용하여, 건조발효사료화, 혼합발효사료화, 혼합 및 건조발효의 병합방법 또는 액상발효사료화 방법이 모두 사용될 수 있다. 건조 발효사료화 방법은 감량화 효과가 높으나, 건조에 투입하는 에너지 비용이 부담스럽고 영양학적으로도 미완의 상태이기 때문에 그 상태로는 이용이 어렵고, 고수분(40%전후)이기 때문에 포장 및 자동 급이라인의 적용에도 어려운 단점이 있다. 혼합발효사료화 방법은 영양적인 밸런스 유지는 가능하나 일령별로 요구량이 다른 다양한 사료를 제조하기 어려운 단점이 있다. 혼합 및 건조발효의 병합 형태는 영양적인 밸런스 유지와 수분 조절이 용이하나, 건조하기 위해 투입되는 에너지 비용과 수분조절제 비용이 포함되어 고비용인 단점이 있다. 수분함량이 85% 전후로 제조되는 액상발효사료는 제조비용이 여타 방법보다 많이 저렴하다는 이점이 있음에도 불구하고 유통의 어려운 단점이 있다.Fermentation feed using food waste using the new strain Bacillus permus BC9 KCCM 10865P, dry fermentation feed, mixed fermentation feed, mixing and dry fermentation combined method or liquid fermentation feed can be used. Dry fermentation feed method has a high weight reduction effect, but it is difficult to use in the state because the energy cost for drying is inconvenient and nutritionally unfinished state, and because it is high moisture (around 40%), packaging and automatic feeding The application of lines also has a disadvantage. The mixed fermentation feed method is capable of maintaining nutritional balance, but it is difficult to manufacture a variety of feeds having different requirements by age. The combined form of mixing and dry fermentation is easy to maintain nutritional balance and moisture control, but has a disadvantage of high cost, including the energy cost and moisture control agent input to dry. Liquid fermented feed produced at around 85% moisture content has a disadvantage in that it is difficult to distribute despite the advantage that the manufacturing cost is much cheaper than other methods.
본 발명의 또 다른 양태는 바실러스 메가터리움 BC1-1 KCCM 10856P(Bacillus megaterium BC1-1 KCCM 10856P), 바실러스 메가터리움 BC1-2 KCCM 10857P(Bacillus megaterium BC1-2 KCCM 10857P), 바실러스 메가터리움 BC2-1 KCCM 10858P(Bacillus megaterium BC2-1 KCCM 10858P), 바실러스 세레우스 BC3 KCCM 10859P(Bacillus cereus BC3 KCCM 10859P), 바실러스 라이케니포미스 BC4 KCCM 10860P(Bacillus licheniformis BC4 KCCM 10860P), 바실러스 세레우스 BC5 KCCM 10861P(Bacillus cereus BC5 KCCM 10861P), 바실러스 스파에리쿠스 BC6 KCCM 10862P(Bacillus sphaericus BC6 KCCM 10862P), 바실러스 클라우시 BC7 KCCM 10863P(Bacillus clausii BC7 KCCM 10863P), 바실러스 라이케니포미스 BC8 KCCM 10864P(Bacillus licheniformis BC8 KCCM 10864P), 바실러스 세레우스 BC10 KCCM 10866P(Bacillus cereus BC10 KCCM 10866P)로 이루어진 그룹에서 선택되는 하나 이상의 바실러스 종과 유효량의 신균주 바실러스 퍼무스 BC9 KCCM 10865P로 구성된 축산폐수를 이용한 유기질 액비 생산 방법 및 음식물 쓰레기를 이용한 가축사료 생산 방법이다.Another embodiment of the present invention is Bacillus megaterium BC1-1 KCCM 10856P ( Bacillus megaterium BC1-1 KCCM 10856P), Bacillus megaterium BC1-2 KCCM 10857P ( Bacillus megaterium BC1-2 KCCM 10857P), Bacillus megaliumium BC2 -1 KCCM 10858P (Bacillus megaterium BC2-1 KCCM 10858P), Bacillus cereus BC3 KCCM 10859P (Bacillus cereus BC3 KCCM 10859P), Bacillus Lai Kenny Po Ms BC4 KCCM 10860P (Bacillus licheniformis BC4 KCCM 10860P), Bacillus cereus BC5 KCCM 10861P (Bacillus cereus BC5 KCCM 10861P), Bacillus spa Erie kusu BC6 KCCM 10862P (Bacillus sphaericus BC6 KCCM 10862P), Bacillus claw when BC7 KCCM 10863P (Bacillus clausii BC7 KCCM 10863P), Bacillus Lai Kenny Po Ms BC8 KCCM 10864P (Bacillus licheniformis BC8 KCCM 10864P), one or more Bacillus species selected from the group consisting of Bacillus cereus BC10 KCCM 10866P and an effective amount of the new strain Bacillus fur. Organic liquid fertilizer production method using livestock wastewater consisting of mousse BC9 KCCM 10865P and livestock feed production method using food waste.
발명의 실시를 위한 형태DETAILED DESCRIPTION OF THE INVENTION
<실시예 1> 바실러스 퍼무스 BC9(<Example 1> Bacillus permus BC9 ( Bacillus firmusBacillus firmus BC9) 균주의 분리 및 동정 BC9) Isolation and Identification of Strains
본 발명에 사용되는 바실러스 퍼무스 BC9(Bacillus firmus BC9) 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. Bacillus permus BC9 ( Bacillus firmus BC9) bacteria used in the present invention was obtained from mushrooms and mycelium masses inhabiting 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 soil sugar, and then incubated at 28-30 ° C.
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.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 BC1-1 KCCM 10856P(Bacillus megaterium BC1-1 KCCM 10856P) 종을 포함하여, 바실러스 메가터리움 BC1-2 KCCM 10857P(Bacillus megaterium BC1-2 KCCM 10857P), 바실러스 메가터리움 BC2-1 KCCM 10858P(Bacillus megaterium BC2-1 KCCM 10858P), 바실러스 세레우스 BC3 KCCM 10859P(Bacillus cereus BC3 KCCM 10859P), 바실러스 라이케니포미스 BC4 KCCM 10860P(Bacillus licheniformis BC4 KCCM 10860P), 바실러스 세레우스 BC5 KCCM 10861P(Bacillus cereus BC5 KCCM 10861P), 바실러스 스파에리쿠스 BC6 KCCM 10862P(Bacillus sphaericus BC6 KCCM 10862P), 바실러스 클라우시 BC7 KCCM 10863P(Bacillus clausii BC7 KCCM 10863P), 바실러스 라이케니포미스 BC8 KCCM 10864P(Bacillus licheniformis BC8 KCCM 10864P), 바실러스 퍼무스 BC9(Bacillus firmus BC9), 바실러스 세레우스 BC10 KCCM 10866P(Bacillus cereus BC10 KCCM 10866P) 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. 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, Bacillus megaterium BC1-2 KCCM 10857P ( Bacillus megaterium BC1-2 KCCM 10857P), including Bacillus megaterium BC1-1 KCCM 10856P, Bacillus megaterium BC2- 1 KCCM 10858P (Bacillus megaterium BC2-1 KCCM 10858P), Bacillus cereus BC3 KCCM 10859P (Bacillus cereus BC3 KCCM 10859P), Bacillus Lai Kenny Po Ms BC4 KCCM 10860P (Bacillus licheniformis BC4 KCCM 10860P), Bacillus cereus BC5 KCCM 10861P ( Bacillus cereus BC5 KCCM 10861P), Bacillus spa Erie kusu BC6 KCCM 10862P (Bacillus sphaericus BC6 KCCM 10862P), Bacillus claw when BC7 KCCM 10863P (Bacillus clausii BC7 KCCM 10863P), Bacillus Lai Kenny Po Ms BC8 KCCM 10864P (Bacillus licheniformis BC8 KCCM 10864P ), Bacillus firmus BC9, Bacillus cereus BC10 KCCM 10866P ( Bacillus cereus BC10 KCCM 10866P) species. It was deposited with the Korean spawn center on April 27, 2007 and each KCCM accession number was assigned.
<실시예 2> 바실러스 퍼무스 BC9(<Example 2> Bacillus permus BC9 ( Bacillus firmusBacillus firmus BC9) 및 바실러스 퍼무스 BC9( BC9) and Bacillus permus BC9 ( Bacillus firmusBacillus firmus BC9)와 다른 바실러스 균의 혼합미생물 조성물에 의한 축산폐수의 악취제거 효과 Deodorization Effect of Livestock Wastewater by Mixed Microbial Composition of BC9) and Other Bacillus
바실러스 퍼무스 BC9에 의한 축산 폐수의 악취 저감 효율을 측정하기 위하여 바실러스 퍼무스 BC9와 바실러스 메가터리움 BC1-1 KCCM 10856P(Bacillus megaterium BC1-1 KCCM 10856P), 바실러스 메가터리움 BC1-2 KCCM 10857P(Bacillus megaterium BC1-2 KCCM 10857P), 바실러스 메가터리움 BC2-1 KCCM 10858P(Bacillus megaterium BC2-1 KCCM 10858P), 바실러스 세레우스 BC3 KCCM 10859P(Bacillus cereus BC3 KCCM 10859P), 바실러스 라이케니포미스 BC4 KCCM 10860P(Bacillus licheniformis BC4 KCCM 10860P), 바실러스 세레우스 BC5 KCCM 10861P(Bacillus cereus BC5 KCCM 10861P), 바실러스 스파에리쿠스 BC6 KCCM 10862P(Bacillus sphaericus BC6 KCCM 10862P), 바실러스 클라우시 BC7 KCCM 10863P(Bacillus clausii BC7 KCCM 10863P), 바실러스 라이케니포미스 BC8 KCCM 10864P(Bacillus licheniformis BC8 KCCM 10864P), 바실러스 퍼무스 BC9(Bacillus firmus BC9), 바실러스 세레우스 BC10 KCCM 10866P(Bacillus cereus BC10 KCCM 10866P) 혼합 미생물군을 일정한 축산 폐수가 들어있는 미생물 반응용기(도 1 참조)에 투입하여 4일 경과 후에 용기 내의 악취를 포집하여 측정하였다. Bacillus megaterium BC1-1 KCCM 10856P, Bacillus megaterium BC1-2 KCCM 10857P ( Bacillus megaterium BC1-1 KCCM 10857P) Bacillus megaterium BC1-2 KCCM 10857P), Bacillus megaterium BC2-1 KCCM 10858P ( Bacillus megaterium BC2-1 KCCM 10858P), Bacillus cereus BC3 KCCM 10859P ( Bacillus cereus BC3 KCCM 10859P) (Bacillus licheniformis BC4 KCCM 10860P), Bacillus cereus BC5 KCCM 10861P (Bacillus cereus BC5 KCCM 10861P), Bacillus spa Erie kusu BC6 KCCM 10862P (Bacillus sphaericus BC6 KCCM 10862P), Bacillus claw when BC7 KCCM 10863P (Bacillus clausii BC7 KCCM 10863P) Bacillus Lai Kenny Po Ms BC8 KCCM 10864P (Bacillus licheniformis BC8 KCCM 10864P), Bacillus buffer mousse BC9 (Bacillus firmus BC9), Bacillus cereus BC10 KCCM 10866P (Bacill us cereus BC10 KCCM 10866P) The mixed microbial group was put into a microbial reaction vessel containing a fixed livestock wastewater (see FIG. 1 ), and 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 유지 시켜 주었다(도 1 참조).The microbial reaction vessel condition of livestock wastewater was set up to maintain the temperature at 37.5 ℃ by using abandonment of constant flow rate and heating band and thermostat for the active activity of 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 퍼무스 BC9 배양액 5ml, 또다른 용기에는 바실러스 메가터리움 BC1-1 KCCM 10856P(Bacillus megaterium BC1-1 KCCM 10856P), 바실러스 메가터리움 BC1-2 KCCM 10857P(Bacillus megaterium BC1-2 KCCM 10857P), 바실러스 메가터리움 BC2-1 KCCM 10858P(Bacillus megaterium BC2-1 KCCM 10858P), 바실러스 세레우스 BC3 KCCM 10859P(Bacillus cereus BC3 KCCM 10859P), 바실러스 라이케니포미스 BC4 KCCM 10860P(Bacillus licheniformis BC4 KCCM 10860P), 바실러스 세레우스 BC5 KCCM 10861P(Bacillus cereus BC5 KCCM 10861P), 바실러스 스파에리쿠스 BC6 KCCM 10862P(Bacillus sphaericus BC6 KCCM 10862P), 바실러스 클라우시 BC7 KCCM 10863P(Bacillus clausii BC7 KCCM 10863P), 바실러스 라이케니포미스 BC8 KCCM 10864P(Bacillus licheniformis BC8 KCCM 10864P), 바실러스 퍼무스 BC9(Bacillus firmus BC9), 바실러스 세레우스 BC10 KCCM 10866P(Bacillus cereus BC10 KCCM 10866P) 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, two 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 permus BC9 culture medium and the other vessel of Bacillus megalithium BC1-1 Bacillus megaterium BC1-1 KCCM 10856P, Bacillus megaterium BC1-2 KCCM 10857P, Bacillus megaterium BC1-2 KCCM 10857P, Bacillus megaterium BC2-1 KCCM 10858P, Bacillus megaterium BC2-1 KCCM 10858P Bacillus cereus BC3 KCCM 10859P ( Bacillus cereus BC3 KCCM 10859P), Bacillus rickeniformis BC4 KCCM 10860P ( Bacillus licheniformis BC4 KCCM 10860P), Bacillus cereus BC5 KCCM 10861P ( Bacillus cereus BC5 KCCM 10861P) 10862P ( Bacillus sphaericus BC6 KCCM 10862P), Bacillus Clauci BC7 KCCM 10863P ( Bacillus clausii BC7 KCCM 10863P), Bacillus rickeniformis BC8 KCCM 10864P ( Bacillus licheniformis BC8 KC CM 10864P), Bacillus firmus BC9 ( Bacillus firmus BC9), Bacillus cereus BC10 KCCM 10866P Was used as a control wastewater.
(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 of 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. The absorbed solution is then transferred to the volumetric flask and the total volume 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 solution for analysis, shake well, mix 5 mL of sodium hypochlorite solution, and leave at 25 to 30 ° C for 1 hour. 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 시험방법2) 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 퍼무스 BC9 및 바실러스 퍼무스 BC9와 다른 바실러스 균의 혼합미생물 조성물을 넣고 4일 배양한 것과 대조폐수만을 넣은 것에 대한 결과를 하기 표 1 에 나타내었다.Bacillus permus BC9 and Bacillus permus BC9 mixed with the microorganism composition of other Bacillus bacteria and the result of the four days culture and only the control waste water is shown in Table 1 below.
표 1TABLE 1
바실러스 퍼무스 BC9는 축산폐수에서 암모니아, 황화수소, 메틸멀캅탄, 다이메틸다이설파이드, 트리메틸아민를 높게 제거하였다. 다이메틸설파이드, 아세트알데히드는 투입 전의 농도와 투입 4일 후 대조시료, 혼합미생물 투입 시료가 거의 유사한 농도로 검출되어 다이메틸설파이드, 아세트알데히드에 대한 제거 효율은 미미한 것으로 보인다.Bacillus permus BC9 had high removal of ammonia, hydrogen sulfide, methyl mercaptan, dimethyl disulfide and trimethylamine from the livestock wastewater. Dimethyl sulfide and acetaldehyde were detected at almost the same concentration as before control, 4 days after the control sample and mixed microbial input sample, and the removal efficiency for dimethyl sulfide and acetaldehyde seems to be insignificant.
<실시예 3> 바실러스 퍼무스 BC9 및 바실러스 퍼무스 BC9와 다른 바실러스 균의 혼합미생물 조성물에 의한 음식물쓰레기의 악취 분석 결과<Example 3> Odor analysis result of food waste by mixed microorganism composition of Bacillus permus BC9 and Bacillus permus BC9 and other Bacillus bacteria
바실러스 퍼무스 BC9에 의한 음식물 쓰레기의 악취 저감 효율을 측정하기 위하여 바실러스 퍼무스 BC9와 바실러스 메가터리움 BC1-1 KCCM 10856P(Bacillus megaterium BC1-1 KCCM 10856P), 바실러스 메가터리움 BC1-2 KCCM 10857P(Bacillus megaterium BC1-2 KCCM 10857P), 바실러스 메가터리움 BC2-1 KCCM 10858P(Bacillus megaterium BC2-1 KCCM 10858P), 바실러스 세레우스 BC3 KCCM 10859P(Bacillus cereus BC3 KCCM 10859P), 바실러스 라이케니포미스 BC4 KCCM 10860P(Bacillus licheniformis BC4 KCCM 10860P), 바실러스 세레우스 BC5 KCCM 10861P(Bacillus cereus BC5 KCCM 10861P), 바실러스 스파에리쿠스 BC6 KCCM 10862P(Bacillus sphaericus BC6 KCCM 10862P), 바실러스 클라우시 BC7 KCCM 10863P(Bacillus clausii BC7 KCCM 10863P), 바실러스 라이케니포미스 BC8 KCCM 10864P(Bacillus licheniformis BC8 KCCM 10864P), 바실러스 퍼무스 BC9(Bacillus firmus BC9), 바실러스 세레우스 BC10 KCCM 10866P(Bacillus cereus BC10 KCCM 10866P) 혼합 미생물군을 일정한 음식물 쓰레기가 들어있는 미생물 반응용기(도 2 참조)에 투입하여 4일 경과 후에 용기 내의 악취를 포집하여 측정하였다.Bacillus permutus BC9 and Bacillus megalium BC1-1 KCCM 10856P ( Bacillus megaterium BC1-1 KCCM 10856P), Bacillus megalium BC1-2 KCCM 10857P Bacillus megaterium BC1-2 KCCM 10857P), Bacillus megaterium BC2-1 KCCM 10858P ( Bacillus megaterium BC2-1 KCCM 10858P), Bacillus cereus BC3 KCCM 10859P ( Bacillus cereus BC3 KCCM 10859P) (Bacillus licheniformis BC4 KCCM 10860P), Bacillus cereus BC5 KCCM 10861P (Bacillus cereus BC5 KCCM 10861P), Bacillus spa Erie kusu BC6 KCCM 10862P (Bacillus sphaericus BC6 KCCM 10862P), Bacillus claw when BC7 KCCM 10863P (Bacillus clausii BC7 KCCM 10863P) Bacillus licheniformis BC8 KCCM 10864P, Bacillus firmus BC9, Bacillus cereus BC10 KCCM 10866P Bacillus cereus BC10 KCCM 10866P) The mixed microbial group was put into a microbial reaction vessel containing a certain food waste (see FIG. 2 ), and after 4 days, 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 유지 시켜 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 퍼무스 BC9 배양액 5ml, 또다른 용기에는 바실러스 메가터리움 BC1-1 KCCM 10856P(Bacillus megaterium BC1-1 KCCM 10856P), 바실러스 메가터리움 BC1-2 KCCM 10857P(Bacillus megaterium BC1-2 KCCM 10857P), 바실러스 메가터리움 BC2-1 KCCM 10858P(Bacillus megaterium BC2-1 KCCM 10858P), 바실러스 세레우스 BC3 KCCM 10859P(Bacillus cereus BC3 KCCM 10859P), 바실러스 라이케니포미스 BC4 KCCM 10860P(Bacillus licheniformis BC4 KCCM 10860P), 바실러스 세레우스 BC5 KCCM 10861P(Bacillus cereus BC5 KCCM 10861P), 바실러스 스파에리쿠스 BC6 KCCM 10862P(Bacillus sphaericus BC6 KCCM 10862P), 바실러스 클라우시 BC7 KCCM 10863P(Bacillus clausii BC7 KCCM 10863P), 바실러스 라이케니포미스 BC8 KCCM 10864P(Bacillus licheniformis BC8 KCCM 10864P), 바실러스 퍼무스 BC9(Bacillus firmus BC9), 바실러스 세레우스 BC10 KCCM 10866P(Bacillus cereus BC10 KCCM 10866P) 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 vessel of 5 ml of Bacillus permus BC9 culture medium and the other vessel of Bacillus megacium BC1-1 KCCM 10856P ( Bacillus megaterium BC1-1 KCCM 10856P), Bacillus megaterium BC1-2 KCCM 10857P ( Bacillus megaterium BC1-2 KCCM 10857P), Bacillus megaterium BC2-1 KCCM 10858P ( Bacillus megaterium BC2-1 KCCM 10858P), Bacillus serenity mouse BC3 KCCM 10859P (Bacillus cereus BC3 KCCM 10859P), Bacillus Lai Kenny Po Ms BC4 KCCM 10860P (Bacillus licheniformis BC4 KCCM 10860P), Bacillus cereus BC5 KCCM 10861P (Bacillus cereus BC5 KCCM 10861P), Bacillus spa Erie kusu BC6 KCCM 10862P ( Bacillus sphaericus BC6 KCCM 10862P), Bacillus Klaus BC7 KCCM 10863P ( Bacillus clausii BC7 KCCM 10863P), Bacillus rickeniformis BC8 KCCM 10864P ( Bacillus licheniformis B C8 KCCM 10864P), Bacillus firmus BC9, Bacillus cereus BC10 KCCM 10866P 5 ml of waste was added and used as a control.
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>
표 2Table 2
바실러스 퍼무스 BC9는 음식물 쓰레기에서 암모니아, 황화수소, 메틸멀캅탄,다이메틸다이설파이드, 트리메틸아민을 높게 제거하였다. 다만, 다이메틸설파이드, 아세트알데히드는 투입 전의 농도와 투입 4일 후 대조시료, 혼합미생물 투입 시료가 거의 유사한 농도로 검출되어 다이메틸설파이드, 아세트알데히드에 대한 제거 효율은 미미한 것으로 보인다.Bacillus permus BC9 removes ammonia, hydrogen sulfide, methyl mercaptan, dimethyl disulfide and trimethylamine from food waste. However, dimethyl sulfide and acetaldehyde were detected at almost the same concentration as the control sample and the mixed microbial input sample after 4 days after the addition, and the removal efficiency of dimethyl sulfide and acetaldehyde seems to be insignificant.
<실시예 4> 축산폐수 및 음식물쓰레기의 중금속 저감효과 분석<Example 4> Heavy Metal Reduction Effect of Livestock Wastewater and Food Waste
바실러스 퍼무스 BC9에 의한 축산폐수 및 음식물 쓰레기에서의 중금속 저감 효과를 측정하기 위하여 11개의 미생물 반응 용기에 음식물쓰레기 각각 2리터씩 일정량 주입한 후 1개의 용기에는 바실러스 퍼무스 BC9 배양액 5ml, 또다른 용기에는 바실러스 메가터리움 BC1-1 KCCM 10856P(Bacillus megaterium BC1-1 KCCM 10856P), 바실러스 메가터리움 BC1-2 KCCM 10857P(Bacillus megaterium BC1-2 KCCM 10857P), 바실러스 메가터리움 BC2-1 KCCM 10858P(Bacillus megaterium BC2-1 KCCM 10858P), 바실러스 세레우스 BC3 KCCM 10859P(Bacillus cereus BC3 KCCM 10859P), 바실러스 라이케니포미스 BC4 KCCM 10860P(Bacillus licheniformis BC4 KCCM 10860P), 바실러스 세레우스 BC5 KCCM 10861P(Bacillus cereus BC5 KCCM 10861P), 바실러스 스파에리쿠스 BC6 KCCM 10862P(Bacillus sphaericus BC6 KCCM 10862P), 바실러스 클라우시 BC7 KCCM 10863P(Bacillus clausii BC7 KCCM 10863P), 바실러스 라이케니포미스 BC8 KCCM 10864P(Bacillus licheniformis BC8 KCCM 10864P), 바실러스 퍼무스 BC9(Bacillus firmus BC9), 바실러스 세레우스 BC10 KCCM 10866P(Bacillus cereus BC10 KCCM 10866P) 배양액이 균등하게 혼합된 혼합미생물 5ml투입, 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.In order to measure the effect of reducing heavy metals in livestock wastewater and food waste by Bacillus permus BC9, 2 liters of food waste were injected into each of 11 microbial reaction vessels, and then one vessel was 5 ml of Bacillus permus BC9 culture solution and another vessel. Bacillus megaterium BC1-1 KCCM 10856P ( Bacillus megaterium BC1-1 KCCM 10856P), Bacillus megaterium BC1-2 KCCM 10857P ( Bacillus megaterium BC1-2 KCCM 10857P), Bacillus megalium BC2-1 KCCM 10858P ( Bacillus megaterium BC2-1 KCCM 10858P), Bacillus cereus BC3 KCCM 10859P (Bacillus cereus BC3 KCCM 10859P), Bacillus Lai Kenny Po Ms BC4 KCCM 10860P (Bacillus licheniformis BC4 KCCM 10860P), Bacillus cereus BC5 KCCM 10861P (Bacillus cereus BC5 KCCM 10861P ), Bacillus spa Erie kusu BC6 KCCM 10862P (Bacillus sphaericus BC6 KCCM 10862P), Bacillus claw when BC7 KCCM 10863P (Bacillus clausii BC7 KCCM 10863P), Basil Srei Kenny Po Ms BC8 KCCM 10864P (Bacillus licheniformis BC8 KCCM 10864P), Bacillus buffer mousse BC9 (Bacillus firmus BC9), Bacillus cereus BC10 KCCM 10866P (Bacillus cereus BC10 KCCM 10866P) culture solution is evenly mixed mixture microorganism 5ml input, One container was used as a control by putting 5ml of food waste without adding microorganisms. 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, hot distilled water is added and filtered using a No. 6 filter paper to a 50 or 100 ml mass flask, followed by rinsing the digestion flask with hot distilled water and thoroughly filtered using a policymen.
(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.
표 3TABLE 3
표 4Table 4
바실러스 퍼무스 BC9는 축산폐수에서 카드뮴, 크롬, 구리, 아연, 납을 높게 제거하였다. 또한, 음식물쓰레기에서 카드뮴, 구리, 아연을 높게 제거하였다. 다만, 음식물쓰레기에서 크롬, 납의 양은 증가되는 것으로 나타나, 축산폐수와는 다른 결과를 보여, 바실러스 퍼무스 BC9에 의한 크롬, 납의 제거효과는 불확실하였다.Bacillus permus BC9 removed cadmium, chromium, copper, zinc and lead from livestock wastewater. In addition, cadmium, copper, and zinc were removed from food waste. However, the amount of chromium and lead was increased in food waste, showing different results from the livestock wastewater, and the effect of removing chromium and lead by Bacillus permus BC9 was uncertain.
본 발명에 따른 바실러스 퍼무스 BC9 KCCM 10865P는 축산폐수 및 음식물쓰레기에서 악취제거 효과가 뛰어났으며, 종래에 사용되는 미생물혼합제(이하 EM이라 칭함)이 악취가 제거되는데 대략 30일의 시간이 소요되는 점(데이타 미첨부)과 비교하여 볼 때 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 permus BC9 KCCM 10865P according to the present invention was excellent in removing odor from livestock wastewater and food waste, and it takes about 30 days to remove the odor of the conventional microbial mixture (hereinafter referred to as EM) Compared to the point (without data), the odor is removed at a very high speed, and the odor removal ability is excellent. 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.
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KR20030023231A (en) * | 2001-09-12 | 2003-03-19 | 최형규 | The composition for composting and deodorizing of animal wastes |
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KR101551699B1 (en) | 2013-12-04 | 2015-09-09 | 경상대학교산학협력단 | Novel bacterial strains used for feeding water flea and clearing culture solution and uses thereof |
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