농법과 계절에 따른 탈질세균의 군집 구조와 탈질율 비교

윤준범,박경량,Yoon, Jun-Beom,Park, Kyeong Ryang 한국미생물학회 2017 미생물학회지 Vol.53 No.1

농법에 따른 토양성분과 $N_2O$ 발생량, 탈질세균 수, 탈질세균의 군집 구조와 T-RFLP 패턴을 계절별로 조사하였다. 토양성분 분석결과 총 탄소량과 총 유기탄소량은 유기농법에서 각각 1.57%, 1.28%, 무농약 농법은 1.52%, 1.24%, 관행농법은 1.40%, 0.95%로 친환경농법에서 유기 탄소량이 비교적 높게 나타났다. $N_2O$ 발생량은 5월과 11월 토양이 높았지만 속도는 8월 토양이 빨랐다. 탈질세균 수는 유기농토양은 평균 $1.32{\times}10^4MPN/g$,무농약 토양은 평균 $1.17{\times}10^4MPN/g$, 관행농 토양은 평균 $6.29{\times}10^3MPN/g$으로 친환경농법 토양이 관행농법 토양에 비해 탈질세균 수가 많은 것을 확인하였다. 계통수 분석 결과, 전체 10개 Cluster 중 유기농법 토양이 6개의 Cluster에 분포되어 친환경 농법 토양이 다양한 군집을 갖는 것을 확인하였다. T-RFLP 패턴의 PCA profile 분석 결과, 유기농법은 넓은 분포를, 관행농법은 좁은 범위의 분포를 나타내고, 무농약농법은 유기농법과 관행농법의 중간에 분포하는 것으로 확인되었다. 따라서 계절과 농법에 따라 탈질세균의 분포와 군집구조가 달라지는 것을 확인하였다. We studied soil composition, $N_2O$ production, a number of denitrifying bacteria, community structure and T-RFLP patterns of denitrifying bacteria dependent on agricultural methods with the change of seasons. Analyses of the soil chemical composition revealed that total carbon and total organic carbon contents were 1.57% and 1.28% in the organic farming soil, 1.52% and 1.24% in the emptiness farming soil, and 1.40% and 0.95% in traditional farming soil, respectively. So, the amount of organic carbon was relatively high in the environment friendly farming soils than traditional farming soils. In case of $N_2O$ production, the amount of $N_2O$ production was high in May and November soils, but the rate of $N_2O$ production was fast in August soil. The average number of denitrifying bacteria were $1.32{\times}10^4MPN{\cdot}g^{-1}$ in the organic farming soil, $1.17{\times}10^4MPN{\cdot}g^{-1}$ in the emptiness farming soil, and $6.29{\times}10^3MPN{\cdot}g^{-1}$ in the traditional farming soil. It was confirmed that the environment friendly farming soil have a larger number of denitrifying bacteria than the traditional farming soil. As a result of the phylogenetic analyses, it was confirmed that six clusters were included in organic farming soil among total 10 clusters. And the result of PCA profile distribution of T-RFLP pattern on agricultural methods, the range of distribution showed wide in the organic farming method, relatively narrow in the conventional farming method, and middle in the emptiness farming method. Therefore, we could concluded that the distribution and the community structure of denitrifying bacteria were changed according to the agricultural methods and seasons.

생물 계면활성제를 생산하는 Pseudomonas sp. G314의 특성

심소희,박경량,Shim, So-Hee,Park, Kyeong-Ryang 한국미생물학회 2006 미생물학회지 Vol.42 No.4

Three hundred thirty two bacterial colonies which were able to degrade crude oil were isolated from soil samples that were contaminated with oil in Daejon area. Among them, one bacterial strain was selected for this study based on its low surface tension ability, and this selected bacterial strain was identified as Pseudomonas sp. G314 through physiological-biochemical tests and analysis of its 16S rRNA sequence. Pseudomonas sp. G314 showed a high resistance to antibiotics such as ampicillin, chloramphenicol, spectinomycin, and streptomycin, and heavy metals such as Li, Cr, and Mn. It was found that the optimal pH and temperature for biosurfactant production of Pseudomonas sp. G314 were pH 7.0 and $30^{\circ}C$, respectively. After seven hours of inoculated, the biosurfactant activity reached the maximum, and surface tension of the culture broth was decreased from 72 to 25 dyne/cm. The crude biosurfactant was obtained from the culture broth by acid precipitation, followed by solvent extraction, evaporation and then freeze drying. The CMC (critical micelle concentration) value of the crude biosurfactant was 20 mg/L. 대전일원의 유류오염 지역의 토양으로부터 원유를 단일 탄소원으로 이용하는 총 322균주를 순수분리 하였고, 이중 생물 계면활성제(biosurfactant) 생성능이 가장 우수한 한 균주를 최종 선별하여 형태 및 생리 생화학적 특성을 조사하고 16S rRNA 염기서열을 분석을 통하여 동정한 결과 Pseudomonas sp.로 확인되어 Pseudomonas sp. G314라 명명하였다. 최종 선별된 Pseudomonas sp. G314는 암피실린, 클로람페니콜, 스펙티노마이신, 스트렙토마이신 등의 항생제와 Li, Cr, Mn 등의 중금속에 대해 강한 내성을 갖고 있었고, 최적 온도와 pH는 각각 $30^{\circ}C$와 pH 7.0으로 확인되었다. Pseudomonas sp. G314가 생성하는 생물 계면활성제의 초기 표면장력은 72 dyne/cm이었으나, 배양 7시간 후 부터는 표면장력이 최대 25 dyne/cm까지 감소되었다. Pseudomonas sp. G3l4가 생산하는 생물 계면활성제를 회수하고 농축하기 위해, 산 침전 후에 유기용매로 배양액을 추출하고 이를 감압농축하여 얻은 시료를 crude biosurfactant로 사용하여 CMC (critical micelle concentration)간을 측정한 결과 20 mg/L로 확인되었다.

Salmonella typhimurium의 금속이온에 대한 유전적 반응

정주리,박경량,고상균,박용근,이인수,Jung, Ju-Ri,Park, Kyeong-Ryang,Koh, Sang-Kyun,Park, Yong-Keun,Lee, In-Soo 한국생명과학회 1998 생명과학회지 Vol.8 No.2

Metal ion-induced and it’s regulatory genes were screened in virulent salmonella typhimurium UK1 and tested cross-regulation with various stresses. Using the techniqud of P22-MudJ(Km, lacZ)-directed lacZ operon fusion, LF40 cuiA::MudJ and Lf153 cuiD::MudJ which were induced by copper were selected. cuia and cuiD were determined anaerobic coper inducible and copper tolerance response gene, respectively. Also cuiA and cuiD locus were determined at 81 and 8min, respectively, on salmonella Genetic Map. The two regulators were identified as cuaR, and cudR, which controls cuiA and cuiD, respectively. cuaR, and cudR appeared as negative regulators because the expression of cuiA-lac-Z and cuiD-lacZ were increased. Copper adapted UK1 showed high resistance to H$_{2}$O$_{2}$, but cuiD did not. The product of the cudR locus was responsible for decreasing the tolerance to copper and H$_{2}$O$_{2}$. Furthemore cuiA and cuiD locus were found to be part of a regulon under the control of a trans-acting regulators, rpoS, oxyR and relA. Therefore, the results suggest CTR participate with oxidative stress on Salmonella.

논과 밭 토양의 황산염 환원세균 군집 구조 비교

이중배,박경량,Lee, Jung Bae,Park, Kyeong Ryang 한국미생물학회 2015 미생물학회지 Vol.51 No.1

본 연구는 논과 밭 토양의 황산염 환원세균의 군집구조와 T-RFLP 패턴을 조사한 논문으로, 유기 농법 토양과 관행 농법 토양 그리고 밭 토양 총 3종류의 토양을 8월과 11월에 채집하여 실험하였다. 토양 성분 분석 결과 총 질소, 총 탄소, 총 인의 값은 모든 토양이 비슷하게 나타났고 계절별로는 수분의 함량은 8월에, 총 탄소는 11월에 가장 높게 나타났다. 황산염 환원세균은 초산보다 젖산을 기질로 이용하는 황산염 환원세균이 더 많이 분포하고, 유기 농법 토양에 황산염 환원세균이 가장 많이 분포하는 것으로 나타났다. 각 토양에서 얻은 총 181개 클론으로 계통학적 분석을 한 결과, 대부분의 클론들은 배양 가능한 황산염 환원세균과는 매우 낮은 상동성을 보였으나, 자연계에서 확인되는 클론들과는 90% 이상의 높은 상동성을 나타내었다. T-RFLP 분석 결과 91, 357, 395, 474 bp의 분포가 가장 높았고, 계절에 따라 황산염 환원세균의 군집 구조가 달라지는 것을 확인하였다. The goal of this study was to identify relationships between the composition of sulfate reducing bacterial assemblages and terminal restriction fragment length polymorphism (T-RFLP) patterns in rice paddy and dry farming soils. Samples of organic farming soils, conventional farming soils, and dry field farming soils were collected in August and November. Analyses of the soil chemical composition revealed similar total nitrogen, total carbon and total inorganic phosphorus levels; however, the moisture content and total carbon were higher than in the other soils in both August and November, respectively. Sulfate reducing bacteria utilizing lactic acid were more widely distributed than those that used acetic acid, and the number of sulfate reducing bacteria in organic farming soil was most abundant. Phylogenetic analysis based on 181 clones revealed that most showed low similarity with cultured sulfate reducing bacteria, but more than 90% similarity with an uncultured sulfate reducing bacteria isolated from the environment. T-RFLP analysis revealed that fragments of 91, 357, 395, and 474 bp were most common, and the community structure of sulfate reducing bacteria changed seasonally.

Pseudomonas rhodesiae H5가 생산하는 Parathion Hydrolase의 특성

윤남경,박경량,Yun Nam Kyung,Park Kyeong Ryang 한국미생물학회 2004 미생물학회지 Vol.40 No.3

The parathion hydrolase (OPH) produced by Pseudomonas rhodesiae H5 was purified by ammonium sulfate precipitation, DEAE-Toyopearl 650M ion exchange chromatography and Sephadex gel filtration chromatography. Parathion hydrolase from crude extracts of P. rhodesiae H5 has two components designated as OPH $I_1$ and OPH $I_2$, Optimum pH and temperature of OPH $I_1$and OPH $I_2$ were pH 7.2 and $30^{\circ}C$, and pH 7.6 and $37^{\circ}C$, respectively. The activation energy of OPH $I_1$ for the hydrolysis of parathion was 3.01 ㎉/I, II, III in the temperature range of $4^{\circ}C$ to $30^{\circ}C$, and Michaelis constant ($K_m$) for parathion was 69.2 ${\mu}M$. The activation energy of OPH $I_2$ for the hydrolysis of parathion was 4.07㎉/㏖ in the temperature range of $4^{\circ}C$ to $37^{\circ}C$, and Michaelis constant for parathion was 150.9${\mu}M$. Furthermore OPH $I_1$ was completely inhibited by 1 mM $Ca^2+$, $Cu^2+$, $Mg^2+$, $Ni^2+$, but OPH $I_2$ was less inhibited than OPH $I_1$ by the metals used in this study. Pseudomonas rhodesiae H5가 생성하는 parathion hydrolase를 $(NH_{4})_{2}SO_{4}$침전, DEAE-Toyopearl 650M ion exchange chromatography, Sephadex gel filtration의 단계로 정제한 결과, 이 균주가 생성하는 parathion hydrolase는 두 종류의 효소였으며, 이를 각각 OPH $I_1$과 OPH $I_2$라 명명하였다. OPH $I_1$과 OPH $I_2$의 최적 pH와 온도는 각각 pH7.2, $30^{\circ}C$와 PH7.6, $37^{\circ}C$이었다. OPH $I_1$의 Parathion을 가수분해하는 활성화 에너지는 $4^{\circ}C$-$30^{\circ}C$에서 3.01㎉/㏖ 이었으며 Km값은 69.2${\mu}M$이었다. 반면, OPH $I_2$의 활성화에너지는 $4^{\circ}C$-$37^{\circ}C$에서 4.07kcal/㏖이었으며 Km값은 150.9${\mu}M$이었다. OPH $I_1$은 1 mM의 $Mg^2+$, $Cu^2+$, $Ca^2+$, $Ni^2+$에 의해서 효소활성이 완전 저해되었으나 OPH $I_2$는 저해를 덜 받는 것으로 확인되었다.

유류오염지역에서 분리한 Acinetobacter sp. B2로부터의 Lipase 정제 및 특성

손성화,박경량,Son Seung Hwa,Park Kyeong Ryang 한국미생물학회 2004 미생물학회지 Vol.40 No.4

대전 근교의 유류로 오염된 토양으로부터 유류를 분해하는332개의 세균 콜로니를 분리한 후 이 중 lipase 활성이 우수한 한 균주를 최종 선별하여 생리생화학적 조사와 16S rRNA 염기서열분석 등을 통하여 동정한 결과 Acinetobactor sp. B2로 확인되었다. 최종 선별된 Acinetobactor sp. B2는 trehalose, mannitol을 제외한 다양한 당을 이용하였고, kanamycin, streptomycin, tetracycline, spectinomycin의 항생제에 대해서 약한 내성을, 그리고 Ba, Li, Mn, Al, Cr, Pb 등의 중금속에 대해서는 mg/ml 단위까지 강한 내성을 나타냈고 생장 최적 온도는 $30^{\circ}C$로 확인되었다. Acinetobactor sp. B2에서 정제된 lipase의 분자량은60 kDa이었고, 이 효소의 최적 온도와 PH는 각각 $40^{\circ}C$와 pH 10이었다. 그리고p-nitrophenyl palmitate (pNPP)를 가수분해하는데 필요한 활성에너지는 $4-37{\circ}C$의 범위에서 2.7 kcal/mol 이었고, $60^{\circ}C$ 이상의 온도에서는 불안정한 효소임이 확인되었다. 또 pNPP에 대한 이 효소의 Michaelis constant (Km)와 최대속도상수 $(V_{max})$값은 각각 21.8 ${\mu}M$과 $270.3\;{\mu}M\;min^{-1}mg^{-1}$ 이었고, 이 효소는 $Cd^{2+},\;Co^{2+},\;Fe^{2+},\;Hg^{2+},$ EDTA, 2-mercaptoethanol에 의해 강하게 억제되었다. Three hundreds thirty two bacterial colonies which were able to degrade crude oil were isolated from soil sam­ples that were contaminated with oil in Daejeon area. Among them, one bacterial strain was selected for this study based on its higher oil degrading ability, and this selected bacterial strain was identified as Acinetobactor sp. B2 through physiological-biochemical tests and analysis of its 16S rRNA sequence. Acinetobactor sp. B2 was able to utilize various carbohydrates but did not utilize trehalose and mannitol as a sole carbon source. Acinetobactor sp. B2 showed a weak resistance to antibiotics such as kanamycin, streptomycin, tetracycline and spectinomycin, but showed a high resistance up to mg/ml unit to heavy metals such as Ba, Li, Mn, AI, Cr and Pb. The optimal growth temperature of Acinetobactor sp. B2 was $30^{\circ}C.$ The lipase produced by Acinetobactor sp. B2 was purified by ammonium sulfate precipitation, DEAE-Toyopearl 650M ion exchange chromatography and Sephadex gel filtration chromatography. Its molecular mass was about 60 kDa and condition for the optimal activity was observed at $40^{\circ}C$ and pH 10, respectively. The activation energy of lipase for the hydrolysis of p­nitrophenyl palmitate was 2.7 kcal/mol in the temperature range of 4 to $37^{\circ}C,$ and the enzyme was unstable at the temperature higher than $60^{\circ}C.$ The Michaelis constant $(K_m)\;and\;V_{max}$ for p-nitrophenyl palmitate were 21.8 uM and $270.3\;{\mu}M\;min^{-1}mg^{-1},$ respectively. This enzyme was strongly inhibited by 10 mM $Cd^{2+},\;Co^{2+},\;Fe^{2+},\;Hg^{2+},$ EDTA and 2-Mercaptoethalol.

Aeromonas sp. MN44의 특성과 망간 산화에 관한 연구

구종서,박경량,Koo Jong Seo,Park Kyeong Ryang 한국생명과학회 2005 생명과학회지 Vol.15 No.1

충청남도 목천과 충청북도 오창 근교의 토양으로부터 망간을 산화하는 64 집락을 분리하고 이 중 망간 산화능이 가장 우수한 한 균주를 최종 선별하여 생리, 생화학적 특성을 조사하고, 16S rRNA 염기 서열분석 등을 통하여 동정한 결과 최종 선별된 균주는 Aeromonas sp. MN44로 확인되었다. 최종 선별된 Aeromenas sp. MN44는 lactose를 제외한 여러 당들은 이용하지 못하였으며, 중금속내성은 lithium과 manganese에 대해서는 mg/ml 단위의 높은 농도까지 중금속 내성을 가지고 있었지만 cadmium에는 전혀 내성을 나타내지 않았다. 또 kanamycin, chloramphenicol, ampicillin, tetracycline, spectinomycin등 조사한 모든 항생제에 대해 전혀 내성을 갖지 않았다. Aeromonas sp. MN44가 생성하는 망간산화물질의 최적 pH는 pH 7.4로 확인되었으며, 이 균이 생성하는 망간 산화 factor는 proteinase K와 가열처리에 의해 저해되는 단백질이고, ammonium sulfate 침전과 ion exchange chromatography 그리고 gel filtration의 단계를 통해 부분 정제한 망간 산화 factor의 분자량은 약 113 kDa로 확인되었다. Sixty four bacterial colonies which were able to oxidize the manganese were isolated from soil samples in Mokcheon and Ochang area. Among them, one bacterial strain was selected for this study based on its higher manganese oxidation, and this selected bacterial strain was identified as Aeromonas sp. MN44 through physiological-biochemical test and analysis of its 16s rRNA sequence. Aeromonas sp. MN44 was able to utilize lactose but did not utilize various carbohydrates as a sole carbon source. Aeromonas sp. MN44 showed a very sensitive to antibiotics such as kanamycin, chloramphenicol, ampicillin, tetracycline and spectinomycin, and heavy metal such as cadmium. But this strain showed a high resistance up to mg/ml unit to heavy metals such as lithium and manganese. Optimal manganese oxidation condition of Aeromonas sp. MN44 was pH 7.4 and manganese oxidation activity was inhibited by proteinase K and boiling treatment. So, we concluded that this factor was protein. The manganese oxidizing factor produced by Aeromonas sp. MN44 was partial purified by ammonium sulfate precipitation, DEAE-Toyopearl 650M ion exchange chromatography and Sephadex gel filtration chromatography. Its molecular mass was about 113 kDa.

농법에 따른 메탄생성과 메탄생성 세균의 T-RFLP 패턴

김훈수,조주식,박경량,Kim, Hun-Soo,Cho, Ju-Sik,Park, Kyeong-Ryang 한국미생물학회 2009 미생물학회지 Vol.45 No.1

농법에 따른 토양의 토양 성분과, 메탄생성량, 메탄생성 세균의 수, 그리고 terminal restriction fragment length polymorphism (T-RFLP) 패턴을 계절별로 조사하였다. 조사기간 동안의 대부분 토양 성분은 큰 차이를 나타내지 않았고, 토양내 물 함량은 5월 시료 보다 10월 시료가 높게 나타났다. 그리고 most probable number (MPN) 방법을 이용한 메탄생성 세균 계수에서, 모든 논토양에 메탄생성 세균이 비교적 많이 존재하였으나 수소를 이용하는 메탄생성 세균과 포름산을 이용하는 메탄생성 세균에 비해 아세트산을 이용하는 메탄 세균수가 상대적으로 적은 것을 확인하였다. 또 포름산 이용 실험에서 대부분 토양이 1주부터 빠른 포름산 이용능을 보여, 4주에는 미량만이 검출되었으나, 아세트산을 첨가한 실험에서는 3주까지 아세트산이 증가된 후, 그 이후에야 아세트산이 이용되어 감소됨을 확인하였다. 그리고 수소를 첨가해준 모든 시료에서는 많은 양의 메탄생성이 있음을 확인하였다. 또 Sou96I을 처리한 mcrA 유전자의 T-RFLP 패턴을 분석한 결과, 농법에 따라 토양성분이 크게 차이가 없는 것과 같이 농법과 계절에 의해서도 뚜렷한 차이를 나타내지 않았지만 일부 통계분석 자료는 유의성이 있음을 확인하였다. 따라서 토양 미생물의 군집비교 기법을 미생물학적 지표로 활용할 수 있다고 판단된다. We studied soil components, methane production, the number of methanogens, and T-RFLP patterns dependent on agricultural methods with the change of seasons. There is no regular increase or decrease tendency of the most soil components followed by sampling period. And the water content in soil was higher in October than May. Also a lot of methanogens existed in soil, and acetotrophs were relatively of smaller number than hydogenotrophs and formate utilizing methanogens using MPN (most probable number) enumeration. In the experiment using the formate, it was used from the first week, and only a minute amount was detecte after four weeks. However in the acetate, it was increased until the third week, and after that was consumed. And there was higher methane production for all soil samples which administered with the hydrogen spike. The activity of methanogens was higher in the organic and low-agrichemical agricultural method samples, and the organic agricultural method had high methanogen activity among the other samples. A result of T-RFLP pattern of mcrA gene digested with Sau96I, methanogen community have a little relation with agricultural methods and seasons. This results also agreed to no critical difference the soil components dependent on agricultural methods, but some analytical data have a positive relationship with a agricultural methods. Therefor we could concluded that the comparison study of community for soil bacteria sufficiently could be useful for the microbiological indicator.

논과 밭 토양의 메탄생성과 메탄생성세균의 군집 비교

김묘선,김주환,박경량,Kim, Myo-Sun,Kim, Joo-Hwan,Park, Kyeong-Ryang 한국미생물학회 2010 미생물학회지 Vol.46 No.4

여름과 가을의 논과 밭 토양의 토양성분과 메탄 생성, 메탄 생성세균의 분포, 그리고 메탄생성세균의 군집구조를 조사하였다. 토양성분을 분석한 결과, 전체적으로 계절에 따라 큰 변화는 나타나지 않았다. 메탄생성세균의 분포조사에서 밭 토양보다 유기농법과 무농약농법을 사용하는 논 토양에 메탄생성세균이 더 많이 존재하였고, 수소와 포름산을 이용하는 메탄생성세균에 비해 아세트산을 이용하는 메탄생성세균 수는 상대적으로 적은 것으로 확인되었다. 메탄생성 실험에서 아세트산을 첨가한 경우 배양 2주까지 메탄생성이 증가되었고, 포름산과 수소를 첨가한 경우 배양 7주까지 메탄생성 양이 증가되었다. mcrA 유전자를 이용한 계통학적 분석에서 논에는 다양한 메탄 생성세균의 cluster가 분포하는 반면, 밭에는 일부 cluster에 집중되어 분포함을 확인하였다. The purpose of this study is to investigate the soil compositions, methane production, the number of methanogens, and the community structure of methanogens in rice paddy soils and dry field farming soils in the summer and autumn seasons. As a result of the analysis of soil compositions, any regular tendency to increase or decrease has not been found in most soil samples due to the change of seasons. It has also been found that more methanogens exist in the rice paddy soil that utilize organic farming practices and emptiness farming practices than in the dry field farming soil. The fewer numbers of methanogens utilizing the acetate have been found than those of the methanogens utilizing the hydrogen or the formate. In an experiment of methane production, the methanes increased for two weeks when the acetate was added, but they continued to increase for seven weeks more when the formate and the hydrogen were added. In the phylogenetic analysis using the mcrA gene, the methanogens had diverse clusters in the rice paddy soil, whereas the methanogens were concentrated only in a few clusters in the dry field farming soil.

Pseudomonas sp. G314가 생산하는 생물 계면활성제의 특성

So Hee Shim(심소희),Kyeong Ryang Park(박경량) 한국생명과학회 2012 생명과학회지 Vol.22 No.2

대전일원의 유류오염 지역의 토양에서 분리된, 생물 계면활성제 생성능이 우수한 Pseudomonas sp. G314균주 [23]가 생산하는 생물 계면활성제의 특성을 조사하고 그 성분을 확인하였다. Pseudomonas sp. G314가 생산하는 생물 계면활성제는 상온에서 10일 보관 후에도 26.2 dyne/㎝ 정도의 표면장력을 유지해 냉장 보관한 계면활성제와 비슷하게 안정하였고, 5 l 발효조를 이용한 배양에서 회분배양의 25 dyne/㎝ 보다는 약간 높은 27 dyne/㎝ 정도의 계면활성제를 생산해 대량 배양 할 수 있음을 확인하였다. 또 이 계면활성제는 acetone과 methanol에 잘용해 되고 benzene과 toluene에 약하게 용해되어 glycolipid 계열의 생물계면활성제임이 추정되었고[23], 이를 silica gel column을 통해 용출하고, TLC로 전개하여 확인된 Rf 0.58인 spot이 bial’s reagent와 rhodamine 6G에서 양성반응을 나타내 Pseudomonas sp. G314가 생산하는 생물 계면활성제는 탄수화물과 지질이 함유된 glycolipid 계열의 생물 계면활성제임을 확인하였다. The purpose of this paper is to analyze the characteristics and chemical components of biosurfactant produced by Pseudomonas sp. G314. Pseudomonas sp. G314 was isolated from soil samples which were contaminated with oil in Daejon area. As such, it produced quality biosurfactant [23]. One type of biosurfactant was kept in a refrigerator, whereas another type of biosurfactant was kept in room temperature. The surface tension activities were then compared. As a result, the biosurfactant from Pseudomonas sp. G314 that was kept at room temperature was stable for 10 days, showing 26.2 dyne/㎝ of surface tension activity. This result was found to be similar to that of the refrigerator storage. The surface tension of batch culture was 25 dyne/㎝, but the culture in the 5 l fermentor was 27 dyne/㎝. Therefore, it can be suggested that the large-scale culture is feasible via the fermentor. Biosurfactant from Pseudomonas sp. G314 was estimated to be a kind of glycolipid because it dissolved in acetone and methanol much better than in benzene and toluene [23]. A spot was detected through the elution of silica gel column and the spread of TLC, and the Rf value was 0.58. This spot has a positive reaction with Bail’s reagent and rhodamine 6G. Hence, we can conclude that biosurfactant from Pseudomons sp. G314 was a glycolipid containing carbohydrate and lipid.