• 한국미생물·생명공학회지
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• 제27권2호
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• pp.159-165
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• 1999

Pseudomonas aeruginosa JRT-4 strain was used as a biosurfactant-producing microorganism in this study. It was one of the microorganisms isolated from the sewage sludge, the main and branch streams of Han river. The surface tension of the culture broth of P. aeruginosa JRT-4 decreased to 30mN/m. The crude biosurfactant was obtained from the culture broth by acid precipitation, solvent extraction, evaporation, and freeze drying. The CMC value of the crude biosurfactant was 0.006%(w/v). From analysis of the chemical structure of biosurfactant, it was determined as rhamnolipid 1 and 3 structures by FAB mass spectrometer. In the washing test for artificially contaminated textiles, the biosurfactant showed better bleachness than the two chemically synthesized surfactant, LAS and SLES. Finally, the biodegradation and ecotoxidolorical tests showed that the biosurfactant was readily biodegradable in the environment and a mild material for microorganisms and green algae.

• 한국미생물·생명공학회지
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• 제22권1호
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• pp.92-98
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• 1994

The bacteria which secrete surface-active agent and decrease the surface tension of culture broth were isolated from soil samples. Among them, biosurfactant producing strain KK-7 was selected and emulsification was also detected. The KK-7 produced biosurfactant not only lipid but also glucose by using carbon source. Taxonomical characterization tests have demostrated the strain KK-7 to be Pseudomonas aeruginosa. The media composition of the P. aeruginosa KK-7 for the biosurfactant production was 1% glucose, 0.5% tryptone, 0.2% yeast extract, 0.15% potas sium phosphate mono-dibasic, 0.05% MgSO$_{4}$, initial pH 8.5, at 30$\circ $C for 2 days. In this condition, the concentration of biosurfactant was reached CMC 5 in the culture broth. Surface active material was produced maximum at stationary8 phase, but emulsification power was higher at log phase than stationary phase. It was considered that P. aeruginosa KK-7 produced biosurfactant more than one type having defferent properties and each maximum production time was different. The minimun surface tension of biosurfactant in 50 mM Tris buffer (pH8.0) was 28 dyn/cm, and CMC was 1 g/L.

• 한국식품영양과학회지
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• 제31권3호
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• pp.389-393
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• 2002

청국장에서 분리한 Bacillus subtilis CH-1, Bacillus circul문 K-1과 Bacillus subtilis(natto) N-1 모두 biosurfactant를 생성하며 이 중 Bacillus subtilis CH-1가 가장 큰 생성력을 나타냈다. Biosurfactant를 대량 생산하기 위하여 AM, LM, NB과 TSB 배지중 AM을 기본배지로 선정하여 최적 탄소원과 질소원으로 glucose 2%, soy peptone 0.3%와 무기염을 포함하는 합성배지를 완성하였다. Biosurfactant의 생성은 96시간에 최대를 나타냈으며 이때 배지의 표면장력은 초기값의 약 43% 값을 나타냈다. 한편 배양온도 및 pH는 biosurfactant생산에 크게 영향을 주지 않았으며 pH5.0~8.0범위에서 대체적으로 안정한 생성을 유지하였다. 최적조건에서 배양시 crude biosurfactant 수율은 6 g/L를 얻을수 있다.

• 생명과학회지
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• 제14권2호
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• pp.320-324
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• 2004

본 연구는 biosurfactant를 생산하는 우수한 균주를 얻고자 산지 토양에서 생산능이 우수한 균주 수십 종을 분리하였다. 분리된 균주 중 원유분해능 및 biosurfactant 생성능이 우수한 균주를 선별하여, 형태학적 특성 및 생리 화학적 특성을 조사고, 16S rDNA의 부분적 염기서열 결정을 통하여 Bacillus sp. TBM911-5로 동정하였다. 동정된 균주 배양액에 생산된 biosurfactant에 의해 48시산 정도에서 표면장력이 최저 29mN/m까지 감소되었다. Bacillus sp. TBM 911-5가 생산하는 biosurfactant의 유화활성은 대두유를 기질로 사용하였을 때 최대였으며, 원유에서도 높은 편이였다. 유화안정성은 합성 계면활성제의 Tween 류나 Triton X-100등과 비슷하거나 우수하였으며, 이는 분리된 균이 생산하는 biosurfactant의 계면활성제로서의 산업적 이용 가능성을 보여준다.

• 생명과학회지
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• 제15권5호
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• pp.679-684
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• 2005

원유 분해능이 강력한 균주를 얻고자 덕유산의 토양으로부터 crude oil을 탄소원으로 이용하는 수십 종을 분리하였다. 분리된 균주 중 원유분해능 및 biosurfactant 생성능이 우수한 균주를 선별하여, 형태학적 특성을 관찰하고 165 rDNA sequence와 』gyrA gene sequence를 통하여 Bacillus atrophaeus DYL-130으로 동정하였다. 동정된 균주 배양액의 표면장력은 최저 28 mN/m까지 감소되었다. 또한 Bacillus atrophaeus DYL-130이 생산하는 biosur-factant가 column chromatography에 의하여 분리 되었으며, 분리된 biosurfactant의 toluene 유화능 및 crude biosurfactant의 유화활성과 안정성이 시험 되었다. Crude biosurfactant의 유화활성은 kerosene에서 최대였으며, soybean oil에서도 높은 편이였으나 유화안정성의 경우 soybean oil을 기질로 하였을 경우가 kerosene을 기질로 하였을 때 보다 우수하였다. 또한 crude biosurfactant의 유화안정성을 합성계면활성제와 비교 한 결과 DYL-130이 생산하는 biosurfactant의 유화안정성이 합성계면활성제와 유사하거나 뛰어남을 확인하였다. Column chroma-tography에 의하여 분리된 biosurfactant는 drop-collapsing method에 의하여 surface-activity가 확인 되었으며 또한 toluene에 대한 유화력이 매우 뛰어난 것을 확인 하였다. 따라서 DYL-130에서 추출한 biosurfactant는 합성계면활성제를 대체할 수 있는 환경친화적인 생물 계면활성제로 사용될 수 있는 가능성을 보여주고 있다. 산업적으로 이 물질을 이용하기 위해서 Bacillus atrephaeus DYL-130이 생산하는 biosurfactant에 대한 구조분석과 물리 화학적 특성, 생분해도 및 환경독성 등의 조사를 수행하여야 할 것으로 생각된다.

• KSBB Journal
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• 제17권3호
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• pp.302-306
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• 2002

탈황공정에 많이 이용되는 미생물인 Rhodococcus sp. Strain IGTS8이 hexadecane과 배지 수용액 혼합물에서 생성하는 biosurfactant의 성분분석과 계면물성을 연구하였다. TLC를 이용한 정성적 성분분석 결과 생성되는 biosurfactant는 glucose mycolate와 trehalose monomycolate임을 확인하였고 임계 미셀농도는 pH 6∼6.5와 pH 10∼10.5에서 100 mL의 수용액에 biosurfactant가 0.1∼0.15 g 첨가될 때였으며 유화파괴 속도는 pH가 높을수록 증가하였다.

• Journal of Microbiology and Biotechnology
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• 제33권9호
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• pp.1238-1249
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• 2023

In this study, we sought to investigate the production and optimization of biosurfactants by soil fungi isolated from petroleum oil-contaminated soil in Saudi Arabia. Forty-four fungal isolates were isolated from ten petroleum oil-contaminated soil samples. All isolates were identified using the internal transcribed spacer (ITS) region, and biosurfactant screening showed that thirty-nine of the isolates were positive. Aspergillus niger SA1 was the highest biosurfactant producer, demonstrating surface tension, drop collapsing, oil displacement, and an emulsification index (E₂₄) of 35.8 mN/m, 0.55 cm, 6.7 cm, and 70%, respectively. This isolate was therefore selected for biosurfactant optimization using the Fit Group model. The biosurfactant yield was increased 1.22 times higher than in the nonoptimized medium (8.02 g/l) under conditions of pH 6, temperature 35℃, waste frying oil (5.5 g), agitation rate of 200 rpm, and an incubation period of 7 days. Model significance and fitness analysis had an RMSE score of 0.852 and a p-value of 0.0016. The biosurfactant activities were surface tension (35.8 mN/m), drop collapsing (0.7 cm), oil displacement (4.5 cm), and E₂₄ (65.0%). The time course of biosurfactant production was a growth-associated phase. The main outputs of the mathematical model for biomass yield were Yx/s (1.18), and µ_max (0.0306) for biosurfactant yield was Y_p/s (1.87) and Y_p/x (2.51); for waste frying oil consumption the So was 55 g/l, and Ke was 2.56. To verify the model's accuracy, percentage errors between biomass and biosurfactant yields were determined by experimental work and calculated using model equations. The average error of biomass yield was 2.68%, and the average error percentage of biosurfactant yield was 3.39%.

• KSBB Journal
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• 제13권5호
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• pp.511-517
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• 1998

Temperature and pH conditions were studied for an effective biosurfactant production by Pseudomonas aeruginosa YPJ-80. Efficient methods of biosurfactant separation were also investigated. pH-uncontrolled experiments at 35$^{\circ}C$ and an initial pH of 8 resulted in the best cell growth (3.6 g/L) and biosurfactant production (0.073 g biosurfactant/g cell). Biosurfactant separation was most efficient using solvent extraction with chloroform/methanol (2:1 vol%) followed by acidification using 1N HCl.

• 한국미생물·생명공학회지
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• 제23권2호
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• pp.229-235
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• 1995

The surface tension-decreasing biosurfactant was purified from Rhodotorula muciloginosa G-1. The purification procedure was the solvent extraction of culture broth. To ensure complete extraction, the sample was extracted twice with equal volume of ethylacetate. The crude solution was washed with n-hexane to remove unconsumed soybean oil. The crude sample of biosurfactant was applied to Silica gel column chromatography equilibrated with chloroform, and eluted with chloroform : methanol gradient. Serveral solvent system was used to developed the thin layer chromatography (TLC). The purified biosurfactant sample gave one spot (Rf 0.78). It was estimated that biosurfactant was glycolipid about having M.W.1,500 with standard of polyethyleneglycol by Sephadex LH-20 column chromatography.

• 한국미생물·생명공학회지
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• 제22권6호
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• pp.593-598
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• 1994

A bacterium KL-57 which exhibited biosurfactant activity was isolated. This bacterium was identified as Bacillus subtilis. The biosurfactant-producing gene of B. subtilis KL-57 was cloned into R subtilis MI113 by using plasmid pTB523. The plasmid DNA from the clone was found to carry a 18 kb PstI insert. The biosurfactant-producing gene was cleaved into 4 fragments by SmaI, 3 fragments by PvulI or EcoRl, 4 fragments by PvulI and EcoRI double digestion, 5 fragments by AccI, and 2 fragments by KpnI, HindIII or BamHI. By subcloning the 18 kb Pstl insert, a 2.3 kb EcoRl fragment conferred the biosurfactant producing activity on B. subtilis cells. The 2.3 kb had one HindIII cleave site. But Two fragments, which corresponds HindIII/EcoRl termini, exhibited no biosurfactant activity.