• 한국환경보건학회지
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• 제26권3호
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• pp.1-10
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• 2000

Although microorganisms play an important role in composting process, researches on the effective microorganism (EM) in composting process are rarely reported. In this study, three stages of work performed 1) investigation of composting facility 2) sample collection and isolation of the EM 3) lab-scale composting by seeding the isolated EM. For this, purpose, physical, chemical, and biological characteristics, such as temperature, moisture content, organic matter, pH population of microorganism, etc., were investigated during the process and the results were compared with those of a control(non-seeded). In the composting facility, the most active degradation of food waste was suppsed to occur in 10-11th day of composting by observing the temperature and CO2 emission. The population of bacterial and fungi was highest in thermophilic stage. Meanwhile that of actinomycetes and yeast was relatively uniform during the process. In the lab-scale test, the thermophilic stage was maintained longer(more than 9 days in 17 days of experiment) in the seeded which was favored for the high reduction of organic matter and moisture. Reduction rates of lipids and salinity were also increased in the seeded. It confirmed these results that the population of microorganisms in the seeded was observed higher in several orders than the control. However, pH of the seeded was maintained as low as about 4.5 throughout the experiment except the final period of the process. Meanwhile, pH of the control rose in the early stage of the experiment. This was probably due that the seeded microorganisms collected from the composting facility was adapted to the low pH environment.

• 한국축산시설환경학회지
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• 제10권2호
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• pp.111-118
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• 2004

본 연구는 양돈 분뇨를 고온호기산화장치(TAO)를 이용하여 처리하였을 때, 고온 미생물 의 첨가에 의한 온도 상승과 시스템의 내부 미생물 변화 그리고 유해 미생물의 불활성화에 대하여 연구하였다. 실험은 총 용량 $18 m^3(3.0\times2.5\times2.4 m)$의 반응기에 양돈 분뇨 $6 m^3$을 투입하고 5$\~$7일간 운전하였다. 대조구는 양돈 분뇨만을 투입하였고 처리구는 6 $\iota$의 고온 미생물(Bacillus. sp)을 투입하였다. 반응기의 내부 미생물의 변화를 검토하기 위해서 호기성 중온균, 고온균 그리고 일반 세균을 분석하였다. 또한, 유해 미생물의 불활성화를 검토하기 위하여 E. coil, Salmonella. sp, Crytosporidium parvum, Giardia lamblia를 분석하였다. 대조구와 처리구의 운전기간 동안 반응기 내부의 온도 범위는 $18\~66^{\circ}C$로 $55^{\circ}C$ 이상의 높은 온도를 유지하였다. 미생물 변화에 있어서 대조구의 중온균과 고온균은 $3.1\times10^6\~1.2\times10^2$ CFU/ml, $1.0\times10^4\~8.0\times10^1$ CFU/ml로 감소하였으나 처리구의 경우, 중온균은 $3.0\times10^8\~8.6\times10^5$ CFU/ml로 감소하였으나 고온균은 $2.0\times10^6\~1.2\times10^8$ CFU/ml로 증가하는 경향을 보였다. Salmonella와 Giardia는 처리 전$\cdot$후에 검출되지 않았으며 E. coil와 Crytosporidium은 처리전 양성반응을 나타내었으나 처리 후 불활성화 되었다. 이상의 결과를 통해서, 우리는 TAO system에 고온 미생물을 첨가함으로써 유해한 미생물이 사멸된 액상비료를 생산할 수 있었고 분뇨로부터 기인하는 2차 오염을 방지할 수 있을 것으로 판단된다.

• 한국미생물생명공학회:학술대회논문집
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• 한국미생물생명공학회 1986년도 추계학술대회
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• pp.509.1-509
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• 1986

Recent advances in recombinant DNA techniques have provided a tool for breeding of microorganisms of hyper production. Enzyme production by cloned microorganism has some advantages. They are ⅰ) Enzymes can be produced by a microorganism easily cultured ⅱ) Hyper production. ⅲ) In some cases, such as thermophilic enzyme gene is cloned in a mesophilic bacteria, the enzyme purification procedure can be simplified. One example, production of thermophilic ${\beta}$-galactosidase in B. subtilis will be presented. Bacillus stearothermophilus IAM 11001 produced three ${\beta}$-galactosidases, ${\beta}$-galactosidase I, II and III (${\beta}$-gal-I, II and III). By connecting restriction fragments of the chromosomal DNA to plasmid vector, followed by transformation of Escherichia coli, two ${\beta}$-galactosidase genes (bgaA and bgaB) located close to each other on the chromosome were cloned.

• 한국식품영양학회지
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• 제13권2호
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• pp.176-180
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• 2000

Microbial protease has been interesting due to the biological roles in the producing microorganism. A thermophilic Actinomyces produing protease was isolated from soil. The optimal medium composition and culture conditions for maximum protease production was as follows 0.5% soluble starch, 0.5% yeast extract. 0.1% K2HPO4, 0.05% CaCl2, initial pH 8.0 at 50$^{\circ}C$ for 48hours. The protease was purified by the procedure of ammonium sulfate precipitation, anion exchange chromatography(LC), DEAE high performance liquid chromatography and GPC HPLC. The purification fold of the purified enzyme was increased about 22.6. The optimal pH and temperature for reaction of the purified enzyme were 7.5 and 60$^{\circ}C$. The purified enzyme was stable for the pH range from 6.0 to 8.5, but was unstable when treated at 80$^{\circ}C$ for 10 minutes. The activity of the enzyme was inhibited by Ag+ and Cu2+.

• Journal of Microbiology and Biotechnology
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• 제9권5호
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• pp.646-649
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• 1999

A thermophilic bacterium producing D-stereospecific dipeptidase was isolated from Korean soil samples. The enzyme hydrolyzed the peptide bond between D-alanyl-D-alanine (D-Ala-D-Ala). The isolated bacterial strain was rod shaped, gram-positive, motile, and formed an endospore. Morphological and physiological characteristics suggested this microorganism a thermophilic Bacillus species, and was named as Bacillus sp. BCS-l. The production of D-stereospecific dipeptidase was growth-associated and optimal at $55^{\circ}C$. The enzyme was applied for the synthesis of D-amino acid-containing peptide, N-benzyloxycarbonyl-L-aspartyl-D-alanine benzyl ester (Z-L-Asp-D-AlaOBzl), as a model reaction. A thermodynamically controlled synthesis of Z-L-Asp-D-AlaOBzl was achieved in an organic solvent.

• 한국폐기물자원순환학회지
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• 제34권2호
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• pp.140-147
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• 2017

This study was conducted to evaluate the effects of physical characteristics. Twelve specific odorous compounds and various sources of bacteria were tested via treatment of food waste using an ultra-thermophilic aerobic composting process. Food waste was mixed with seed material and operated for 47 days. During composting, the temperature was maintained at $80-90^{\circ}C$. The variations in $O_2$, $CO_2$ and $NH_3$ production suggested typical microorganism-driven organic decomposition patterns. After composting, the concentrations of 12 specific odorous compounds other than ammonia did not exceed the allowable exhaust limits for odor. After composting, thermophiles represented 50% of all bacteria. After composting, the percentage of thermophile bacterial increased by 15%. Therefore, both stable composting operation and economic benefit can be expected when an ultra-thermophilic composting process is applied to food waste.

• Journal of Microbiology and Biotechnology
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• 제18권9호
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• pp.1544-1549
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• 2008

MhMTS and MhMTH are trehalose ($\alpha$-D-glucopyranosyl-[1,1]-$\alpha$-D-glucopyranose) biosynthesis genes of the thermophilic microorganism Metallosphaera hakonensis, and encode a maltooligosyltrehalose synthase (MhMTS) and a maltooligosyltrehalose trehalohydrolase (MhMTH), respectively. In this study, the two genes were fused in-frame in a recombinant DNA, and expressed in Escherichia coli to produce a bifunctional fusion enzyme, MhMTSH. Similar to the two-step reactions with MhMTS and MhMTH, the fusion enzyme catalyzed the sequential reactions on maltopentaose, maltotriosyltrehalose formation, and following hydrolysis, producing trehalose and maltotriose. Optimum conditions for the fusion enzyme-catalyzed trehalose synthesis were around $70^{\circ}C$ and pH 5.0-6.0. The MhMTSH fusion enzyme exhibited a high degree of thermostability, retaining 80% of the activity when pre-incubated at $70^{\circ}C$ for 48 h. The stability was gradually abolished by incubating the fusion enzyme at above $80^{\circ}C$. The MhMTSH fusion enzyme was active on various sizes of maltooligosaccharides, extending its substrate specificity to soluble starch, the most abundant natural source of trehalose production.

• 산업식품공학
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• 제14권1호
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• pp.7-13
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• 2010

호열성 미생물을 검토하기 위하여 전국 각지로부터 토양과 두엄을 채취하여 그로부터 호열성 미생물을 분리하였다. 토양과 두엄으로부터 분리된 호열성 미생물 1250여 균주를 선별하였고, 이들을 대상으로 미생물이 생산하는 효소 활성을 검토하여 호열성 전분 분해 효소를 생산하는 1주의 미생물을 확인하였다. 확인된 1주의 미생물을 strain 2719라 명명하였다. Strain 2719 균주는 형태학적으로 gram 양성 간균의 특징을 나타냈고, 균주의 표면은 매끄럽지 않았으며, 비교적 다양한 길이를 가지고 있었다. 또한 다른 gram 양성 간균들에 비해서 많은 수의 균사들이 각 균주들 사이에 복잡하게 얽혀있었다. 생화학적 특성을 확인한 결과 catalase 양성, glucose 발효, arabinose 발효, mannitol 발효, casein gelatin starch 가수분해의 특징을 가지고 있었으며, 이는 Bacillus sp.로 추정되었다. 생육 pH의 범위는 pH 6-pH 8범위에서 생육이 가능했으며, 생육 온도의 범위는 50-70${^{\circ}C}$였다. 16S rDNA sequence 분석결과 Bacillus thermoglucosidasius의 16S rDNA와 99.52%가 일치하였으나, sequence의 일부분이 다른 부분이 있고, 생육 특성에서 약간의 차이를 보였다. 또한 gene bank에 등록되어 있는 균주들의 16S rDNA sequence들과 비교하여도 일치하는 균주는 확인되지 않았다. 이와 같은 실험결과에 따라 2719 균주는 기존에 발표되지는 않았으나, Bacillus thermoglucosidasius와 매우 유사한 균주로 판단되어 Bacillus thermoglucosidasius 2719로 명명하였다.

• 한국미생물·생명공학회지
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• 제41권3호
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• pp.278-283
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• 2013

A gram-positive, rod-shaped, spore-forming, motile thermophilic bacterium was isolated from a sterilization oven. The microorganism GWE1, formally named Geobacillus wiegelii identified as a member of the genus Geobacillus. GWE1 grew under aerobic conditions of between $60-80^{\circ}C$ (optimum $670^{\circ}C$), in a pH range of 3.0-8.0 (optimum $pH^{70^{\circ}C}$ 5.8), and between 0 and 2 M NaCl (optimum 0.3 M). The membrane polar lipids were dominated by branched saturated fatty acids, which included as the major constituents; iso-15:0 (13.3%), 16:1(${\omega}7$) (12.8%), 16:0 (28.5%), iso-17:0 (13.5%) and anteiso-17:0 (12.3%). The DNA G+C content was 47.2 mol% (determined by HPLC). The 16S rRNA gene sequence of GWE1 showed a high similarity with Geobacillus caldoxylosilyticus (97%). However, the level of DNA-DNA relatedness was only 58%. These data suggest that GWE1 is probably a novel specie of the genus Geobacillus.

• 한국식품과학회지
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• 제38권3호
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• pp.419-426
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• 2006

전국 각지에서 채집한 토양에서 분리한 25종의 내열성 균주 중 내열성 단백질 기수분해효소 활성을 갖는 균주 strain JE 375를 선별하였다. 본 균주는 gram 양성 간균의 특징을 나타냈으며 Bergey's Manual of Systematic Bacteriology와 Biochemical Tests for Identification of Medical Bacteria에 준하여 생화학적 특성을 검토한 결과 catalase 양성, 포자형성, motility 양성, glucose 발효, mannitol 발효, xylose 산화, hemolysis ${\beta}$균임을 보아 Bacillus sp.으로 추정 되었다. Strain JE 375의 whole cell fatty acid를 gas chromatography로 분석한 결과 $C_{15:0}$ iso 26.17%, $C_{16:0}$ iso 13.01%, $C_{17:0}$ iso 30.19%로 분석되어 Bacillus 계열로 동정되었다. 16S rDNA sequence분석 결과 strain JE 375는 Bacillus caldoxylolyticus와 sequence가 97.6% 일치하는 유사성을 보였으나 부분적으로 sequence의 차이가 있고 gene bank data base상에서 16S rDNA sequence가 일치하는 균주는 검색되지 않았다. 이 같은 실험 결과에 따라 strain JE 375는 기존에 발표되지 않은 새로운 균주로 판단되어 Bacillus sp. JE 375로 명명하였다. Bacillus sp. JE 375은 tryptone 1%, yeast extract 0.5%, NaCl 1%, maltose 1%의 배지조성분과 배양 온도 $65^{\circ}C$에서 20시간 동안 배양하였을 때 최대의 단백질 분해 효소를 생산하였다. Bacillus sp. JE 375로부터 단백질 분해 효소를 acetone으로 침전시키고 DEAE-sepharose column chromatography를 통하여 효소를 정제하여 SDS-PAGE를 통해 확인한 결과 55 kDa 크기의 band를 확인할 수 있었다. 이 효소의 최적 배양 온도는 $65^{\circ}C$이었으며 배지의 최적 pH는 6.5로 나타났다. pH에 대한 안정성은 중성 부근의 pH에서 효소 활성의 안정성이 높게 나타났다. 본 효소의 반응 조건을 검토한 결과 중성 조건에서 안정하였으며, $60^{\circ}C$의 고온에서 활성을 가졌다. 효소 활성은 1 mM $CaCl_2$ 첨가에 의해 증가하였다.