KR20040094568A - A bacteria to stimulate organic waste composting - Google Patents
A bacteria to stimulate organic waste composting Download PDFInfo
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- KR20040094568A KR20040094568A KR1020030028432A KR20030028432A KR20040094568A KR 20040094568 A KR20040094568 A KR 20040094568A KR 1020030028432 A KR1020030028432 A KR 1020030028432A KR 20030028432 A KR20030028432 A KR 20030028432A KR 20040094568 A KR20040094568 A KR 20040094568A
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Abstract
Description
우리나라의 생활폐기물의 발생량은 44,583톤(98년 기준)이며 이중 음식물 쓰레기의 발생량은 11,062톤으로 약 24.8%를 차지하고 있다. 특히, 음식물 쓰레기는 수분함량이 높고 쉽게 부패되므로 매립이나 소각에 어려운 문제점을 안고 있다. 음식물쓰레기의 재활용 비율은 '98년의 22%에서 2000년에는 43%로 증가하였으며 이중에서 퇴비화로 처리되는 재활용 비율은 35.3%에 달하고 있다(2001 환경산업총람).The amount of domestic waste generated in Korea is 44,583 tons (as of 1998), and the amount of food waste generated is 11,062 tons, accounting for 24.8%. In particular, food waste has a high moisture content and easily decayed, which makes it difficult to landfill or incinerate. The recycling rate of food waste increased from 22% in 1998 to 43% in 2000, of which 35.3% was recycled through composting (2001 Environmental Industry Show).
쓰레기의 처리방법 중 퇴비화는 유기성 폐기물의 적용에 가장 실용적인 처리방법으로 평가받고 있다. 퇴비화는 미생물에 의한 유기물의 분해과정이며(안정화 단계) 최종적으로는 리그닌과 같은 난분해성 물질을 분해하여 휴믹(humics)을 생성시켜 토양에 공급하는 개념이다. 이와 같이 퇴비화 공정은 미생물을 필요로 하며, 최근 대부분의 퇴비화공장에서는 미생물을 외부에서 첨가하는 방법을 사용하고 있다. 외국에서도 이미 균주 첨가가 각종 쓰레기의 분해 및 퇴비화를 촉진시킨다는 연구결과가 보고되었다(Appl. Environ. Microbiol. 49: 724-726(1985)). 그러나 국내에서는 사용목적에 따라 적합한 종균제용 균주 선발을 위한 체계적인 분리방법과균주의 특성에 따른 연구보고는 미비한 실정이다.Composting is regarded as the most practical method for the application of organic waste. Composting is the process of decomposition of organic matter by microorganisms (stabilization step). Finally, it is a concept of decomposing hardly decomposable substances such as lignin to generate humics and supply them to the soil. As such, the composting process requires microorganisms, and recently, most composting plants use a method of adding microorganisms externally. Overseas studies have also reported that the addition of strains promotes the decomposition and composting of various wastes ( Appl. Environ. Microbiol . 49: 724-726 (1985)). However, in Korea, there are few research reports based on systematic separation methods and strains for the selection of suitable strains for spawn.
본 발명이 이루고자하는 기술적 과제는 음식물 쓰레기의 고속발효에 관여하는 미생물을 1종 분리하고 형태, 생태학적 특성 및 효소활성 등을 조사하므로써 유기성 폐기물의 퇴비화 종균제로 적합한 균주개발의 기초자료로 활용한다.The technical problem to be achieved by the present invention is to isolate one species of microorganisms involved in the high-speed fermentation of food waste and to investigate the form, ecological characteristics and enzyme activity, and to use as a basic data for the development of strains suitable for composting spawn of organic waste.
본 발명은 유기성 폐기물의 퇴비화 촉진을 목적으로 사용 가능한 1종의 미생물을 제공한다. 본 발명에 따른 균주는 음식물 쓰레기의 호기성 발효(퇴비화)장치에서 분리한Bacillus sp. MB23균은 amylase, protease, protease와 lipase 효소활성이 모두 우수하였다.The present invention provides one microorganism that can be used for promoting composting of organic waste. The strain according to the present invention is Bacillus sp . Isolated from aerobic fermentation (composting) device of food waste. MB23 was excellent in amylase, protease, protease and lipase enzyme activities.
이하, 본 발명의 구체적인 구성과 작용을 단계별로 상세히 설명한다.Hereinafter, the specific configuration and operation of the present invention will be described in detail step by step.
균주의 분리Isolation of Strains
음식물 쓰레기의 호기성 발효장치는 안지름 430mm, 높이 700mm, 두께 10mm, 최대용량 801의 아크릴수지로 제작하였고 반응조의 외부는 30mm 두께의 아티론을 단열재로 사용하여 열에너지의 손실을 억제하였다. 수분 조절제는 톱밥과 대패밥을 2:1로 사용하였으며 통기속도는 751/m2·min로 하였고 교반은 1일 1회 수행하였다.The aerobic fermentation system for food waste was made of acrylic resin with inner diameter of 430mm, height of 700mm, thickness of 10mm, and maximum capacity of 801. The outside of the reactor used 30mm thick atyrone as insulation material to suppress the loss of thermal energy. Moisture control agent was used as a 2: 1 sawdust and large rice and aeration rate was 751 / m 2 · min and stirring was carried out once a day.
미생물의 분리는 발효장치내의 시료 5g(습윤중량)을 취하여 증류수 95ml와 섞어 40W에서 1분간 초음파 분쇄기로 균일하게 현탁한후 적절히 희석하여 접종원으로 사용하였다. 분리용 평판배지로서는 일반세균용(Soybean-casein digestmedium), 방선균(Actinomycete isolation agar medium) 그리고 균류용(Potatodextrose)의 3종류(표 1)를 사용하였다. pH는 균류용 배지가 5.6으로 조절된 것을 제외한 방선균 및 일반세균은 7.3으로 조정하였다. 중온성과 고온성 미생물의 계수를 위한 배양온도는 각각 30℃와 55℃로 하였다.The microorganisms were separated by 5 g (wet weight) of the sample in the fermentation apparatus, mixed with 95 ml of distilled water, uniformly suspended in an ultrasonic grinder for 1 minute at 40 W, and then diluted appropriately and used as an inoculum. As the plate for separation, three types (Soybean-casein digestmedium), Actinomycete isolation agar medium, and fungus (Potatodextrose) (Table 1) were used. The pH was adjusted to 7.3 for actinomycetes and general bacteria except that the fungal medium was adjusted to 5.6. The culture temperature for the count of mesophilic and pyrogenic microorganisms was 30 ℃ and 55 ℃, respectively.
균주의 선별 및 생리, 형태학적 관찰Screening, Physiology and Morphological Observation of Strains
중온균의 분리를 위해 30℃, 고온균의 분리를 위해 55℃의 배양온도에서 배양한 후 1차로 37종을 분리하였다. 이중 amylase, protease, cellulase, lipase의 활성이 우수한 고온성 세균 6종, 중온성 세균 7종을 2차로 선별하였다. 표 2 에는 2차로 선별된 고온균 6주, 중온균 7주의 형태학적, 생리학적 특징을 나타내었다. 선별된 모든 고온성 균주는 간균의 형태를 보였으며 포자를 형성하고 카탈라제 양성반응을 보였다. 또한 중온성균의 경우도 MB4와 MB18이 구균의 형태를 지닌 것을 제외하고는 MB10, MB11, MB15-1, MB15-2, MB23 모두 간균이으로 이들 중온균 7종 모두 그람양성이었다. 또한, MB4, MB11과 MB18은 포자를 형성하지 않았으며 MB10, MB15-1, MB15-2, MB23은 포자를 형성하였다. 분리된 모든 중온성 균은 카탈라제 양성반응을 보였다.For the isolation of mesophilic bacteria, 37 species were firstly isolated after incubation at a culture temperature of 55 ° C. for the separation of high temperature bacteria. Among them, 6 thermophilic bacteria and 7 mesophilic bacteria with excellent amylase, protease, cellulase and lipase activity were selected. Table 2 shows the morphological and physiological characteristics of 6 strains of 7 strains of thermophile and 7 strains of mesophilic strains. All of the selected thermophilic strains were in the form of bacillus, forming spores and positive catalase. In the case of mesophilic bacteria, MB10 and MB11, MB15-1, MB15-2, and MB23 were all bacilli, except that MB4 and MB18 had the form of cocci. MB4, MB11, and MB18 did not form spores, and MB10, MB15-1, MB15-2, and MB23 formed spores. All mesophilic isolates were positive for catalase.
2차선별을 거친 균중에서 각 기질에 대한 고른 효소활성을 나타낸 균주, 즉 중온균으로 MB15-1, MB15-2와 MB23, 고온균으로 TB1과 TB9를 3차로 선별하였다. 이렇게 분리된 균주의 동정은 "Bergey's Manual of Systematic Bacteriology"에 준하여 실시하였다. 표 3은 3차 선별을 거친 중온균 3종와 고온균 2종에 대한 탄소원 이용성과 질산염 환원성 등 생리학적 특성을 나타내었다.Among the bacteria that had undergone secondary screening, MB1-1, MB15-2 and MB23 were selected as the mesophilic bacterium, and TB1 and TB9 were selected as the mesophilic bacteria. Identification of the isolated strain was carried out according to "Bergey's Manual of Systematic Bacteriology". Table 3 shows the physiological characteristics such as carbon source availability and nitrate reducibility of three mesophilic and 2 high temperature bacteria.
본 발명에 따른 균주Bacillus sp. MB23의 형태학적 특성을 관찰하기 위해 광학현미경 및 주사현미경(SEM)을 이용하였다. 여기에서 주사현미경에 의한 관찰을 위한 시료의 전처리는 먼저, pH 7.2, 0.1M calcium carolyate 완충액으로 두 번 세척하고 0.1M calcium carolyate용액에 2.5% 농도로 녹인 glutaraldehyde용액으로 pH 4.0에서 72시간동안 고정하였다. 그후 시료를 물과 에탄올 혼합액(50-100%)에서 45분간 단계적으로 탈수과정을 거치고 100% 에탄올에서 16시간 처리하였다. 그 다음 CO2를 이용하여 임계점 건조후 탄소로 코팅하고 Jeol JSM-T300 주사현미경으로 균주의 유기물 분해능력을 20Kv에서 촬영하였다. 그림 1에서는 본 발명에서 분리, 동정한Bacillus sp. MB23의 주사현미경 사진이다.The strain Bacillus sp . Optical and scanning microscope (SEM) were used to observe the morphological characteristics of MB23. Here, pretreatment of the sample for observation by scanning microscope was first washed twice with pH 7.2, 0.1 M calcium carolyate buffer and fixed at pH 4.0 for 72 hours with glutaraldehyde solution dissolved in 2.5% concentration in 0.1 M calcium carolyate solution. . The sample was then dehydrated in 45 minutes in water and ethanol mixture (50-100%) and treated for 16 hours in 100% ethanol. Then, after drying the critical point using CO 2 and coated with carbon, the organic decomposition ability of the strain was photographed at 20Kv by Jeol JSM-T300 scanning microscope. Figure 1 shows Bacillus sp isolated and identified in the present invention. Scanning micrograph of MB23.
[실시예 1]Example 1
Bacillus sp. MB 23의 유기 고분자물질 분해활성 Bacillus sp . Degradation Activity of MB 23 Organic Polymer
본 발명에 따른Bacillus sp. MB 23의 고분자 분해능력을 평가하기 위해 amylase, protease, cellulase와 lipase 효소활성을 측정하였다. 표 4에서는Bacillus sp. MB 23의 효소활성을 1차적으로 분리한 다른 균주들과 비교하여 나타내었다. Bacillus sp according to the present invention. Amylase, protease, cellulase and lipase enzyme activities were measured to evaluate the polymer degradation capacity of MB 23. Table 4 shows Bacillus sp . Enzyme activity of MB 23 was shown in comparison with other strains isolated first.
Amylase 활성측정Amylase Activity Measurement
1%의 soluble starch를 첨가한 nutrient agar 배지를 사용하였으며, 측정방법은 지름이 17cm인 멸균 petri dish에 배지를 부어 굳히고 배지표면을 풍건한 후, 내경이 7mm인 stainless steel cylinder를 올려놓고, LB배지에서 18시간 배양한 배양액 20ul를 cylinder내로 주입한 다음 일정한 시간 배양하고 I2-KI용액을 분무하여 형성된 clear zone의 지름을 측정하였다.A nutrient agar medium containing 1% soluble starch was used.Measurement method was to pour the medium into a sterile petri dish with a diameter of 17cm, harden the medium, dry the surface of the medium, and place a stainless steel cylinder with an internal diameter of 7mm. 20ul of the culture medium incubated at 18 hours was injected into the cylinder, and then cultured for a predetermined time, and the diameter of the clear zone formed by spraying the I 2 -KI solution was measured.
Protease 활성측정Protease Activity Measurement
1%의 skim milk를 첨가한 nutrient agar 배지를 사용하였으며, skim milk가 분해되어 생긴 clear zone의 지름을 측정하였다.A nutrient agar medium containing 1% skim milk was used, and the diameter of the clear zone resulting from the decomposition of skim milk was measured.
Cellulase 활성측정Cellulase Activity Measurement
기질로서 0.5% CM-cellulose를 함유하고 있는 배지를 사용하였으며, 0.1% congored 용액을 가하고 30분간 유지시킨 후, 1M NaCl 용액으로 15분간 세척했을때 생긴 clear zone의 지름을 측정하였다.The medium containing 0.5% CM-cellulose was used as the substrate, and 0.1% congored solution was added and maintained for 30 minutes, and then the diameter of the clear zone generated when washed with 1M NaCl solution for 15 minutes was measured.
Lipase 활성측정Lipase Activity Measurement
LB배지에 0.5%의 tributyrin을 첨가한 배지를 사용하였으며, tributyrin이 분해되어 생긴 clear zone의 지름을 측정하였다.The medium to which 0.5% tributyrin was added to the LB medium was used, and the diameter of the clear zone resulting from degradation of tributyrin was measured.
본 발명은 음식물 쓰레기의 호기성 발효장치에서 퇴비화 촉진용 목적으로 미생물을 분리하고 형태, 생리학적 특성을 검토한 결과Bacillus sp.로 동정되었다. 또한, 이상의 실시예에서 확인되는 바와 같이Bacillus sp. MB23은 유기성 폐기물의 처리에 매우 유용한 균으로 판단된다.The present invention was identified as Bacillus sp . As a result of the separation of microorganisms and the morphology and physiological characteristics of the aerobic fermentation system for food waste in order to promote composting. In addition, Bacillus sp . MB23 is considered a very useful bacterium for the treatment of organic wastes.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20150107004A (en) * | 2014-03-12 | 2015-09-23 | 주식회사 템셀코리아 | Microorganism for decomposing a food waste, a composition comprising the same and the eliminating method of a food waste using the same |
| KR20160003587A (en) * | 2015-12-11 | 2016-01-11 | 주식회사 템셀코리아 | Microorganism for decomposing a food waste, a composition comprising the same and the eliminating method of a food waste using the same |
| KR20190027805A (en) * | 2019-03-08 | 2019-03-15 | 주식회사 템셀코리아 | Composition for decomposing a food waste |
| KR20190028407A (en) * | 2019-03-08 | 2019-03-18 | 주식회사 템셀코리아 | Composition for decomposing a food waste |
| KR20190028406A (en) * | 2019-03-08 | 2019-03-18 | 주식회사 템셀코리아 | Composition for decomposing a food waste |
-
2003
- 2003-05-03 KR KR1020030028432A patent/KR20040094568A/en not_active Application Discontinuation
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20150107004A (en) * | 2014-03-12 | 2015-09-23 | 주식회사 템셀코리아 | Microorganism for decomposing a food waste, a composition comprising the same and the eliminating method of a food waste using the same |
| KR20160003587A (en) * | 2015-12-11 | 2016-01-11 | 주식회사 템셀코리아 | Microorganism for decomposing a food waste, a composition comprising the same and the eliminating method of a food waste using the same |
| KR20190027805A (en) * | 2019-03-08 | 2019-03-15 | 주식회사 템셀코리아 | Composition for decomposing a food waste |
| KR20190028407A (en) * | 2019-03-08 | 2019-03-18 | 주식회사 템셀코리아 | Composition for decomposing a food waste |
| KR20190028406A (en) * | 2019-03-08 | 2019-03-18 | 주식회사 템셀코리아 | Composition for decomposing a food waste |
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