• Journal of Microbiology and Biotechnology
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• 제23권3호
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• pp.390-396
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• 2013

Biosurfactants have versatile properties and potential industrial applications. A new producer, B. subtilis TU2, was isolated from the underground oil-extraction wastewater of Shengli Oilfield, China. Preliminary flask culture showed that the titer of biosurfactant obtained from the broth of TU2 was ~1.5 g/l at 48 h (718 mg/l after purification), with a reduced surface tension of 32.5 mN/m. The critical micelle concentration was measured as 50 mg/l and the surface tension maintained stability in solution with 50 g/l NaCl and 16 g/l $CaCl_2$ after 5 days of incubation at $70^{\circ}C$. FT-IR spectra exhibited the structure information of both glycolipid and lipopeptide. MALDI-TOF-MS analyses confirmed that the biosurfactant produced by B. subtilis TU2 was a blend of glycolipid and lipopeptide, including rhamnolipid, surfactin, and fengycin. The blended biosurfactant showed 86% of oil-washing efficiency and fine emulsification activity on crude oil, suggesting its potential application in enhanced oil recovery.

• Environmental Engineering Research
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• 제16권4호
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• pp.219-225
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• 2011

The objective of this study was to investigate the bacterial adhesion to iron (hydr)oxide-coated sand (IHCS) and aluminum oxidecoated sand (AOCS) in the presence of Tween 20 (nonionic surfactant) and lipopeptide biosurfactant (anionic surfactant) through column experiments. Results show that in the presence of Tween 20, bacterial adhesion to the coated sands was slightly decreased compared to the condition of deionized water; the mass recovery (Mr) increased from 0.491 to 0.550 in IHCS and from 0.279 to 0.380 in AOCS. The bacterial adhesion to the coated sands was greatly reduced in lipopeptide biosurfactant; Mr increased to 0.980 in IHCS and to 0.797 in AOCS. Results indicate that the impact of lipopeptide biosurfactant on bacterial adhesion to metal oxide-coated sands was significantly greater than that of Tween 20. Our results differed from those of the previous report, showing that Tween 20 was the most effective while the biosurfactant was the least effective in the reduction of bacterial adhesion to porous media. This discrepancy could be ascribed to the different surface charges of porous media used in the experiments. This study indicates that lipopeptide biosurfactant can play an important role in enhancing the bacterial transport in geochemically heterogeneous porous media.

• Journal of Microbiology
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• 제43권3호
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• pp.272-276
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• 2005

Bacillus subtilis ATCC 6633 was grown in BHIB medium supplemented with $Mn^{2+}$ for 96 h at $37^{\circ}C$ in a shaker incubator. After removing the microbial biomass, a lipopeptide biosurfactant was extracted from the supernatant. Its structure was established by chemical and spectroscopy methods. The structure was confirmed by physical properties, such as Hydrophile-Lipophile Balance (HLB), surface activity and erythrocyte hemolytic capacity. The critical micelle concentration (cmc) and erythrocyte hemolytic capacity of the biosurfactant were compared to those of surfactants such as SDS, BC (benzalkonium chloride), TTAB (tetradecyltrimethylammonium bromide) and HTAB (hexadecyltrimethylammonium bromide). The maximum hemolytic effect for all surfactants mentioned was observed at concentrations above cmc. The maximum hemolytic effect of synthetic surfactants was more than that of the biosurfactant produced by B. subtilis ATCC 6633. Therefore, biosurfactant would be considered a suitable surface-active agent due to low toxicity to the membrane.

• Journal of Microbiology and Biotechnology
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• 제7권3호
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• pp.180-188
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• 1997

Bacillus subtilis C9 was selected by measuring the oil film-collapsing activity and produced biosurfactant in a medium containing glucose as a sole carbon source. The biosurfactant emulsified hydrocarbons, vegetable oils and crude oil, and lowered the surface tension of culture broth to 28 dyne/cm. A biosurfactant, C9-BS produced by B. subtilis C9 was purified by ultrafiltration, extraction with chloroform and methanol, adsorption chromatography, and preparative reversed phase HPLC. Structural analyses, IR spectroscopy, FAB mass spectroscopy, amino acid composition, and NMR analyses, demonstrated that C9-BS was a lipopeptide comprising a fatty acid tail and peptide moiety. The lipophilic part consisting of $C_{14}\;or\;C_{15}$ hydroxy fatty acid was linked to the hydrophilic peptide part, which contained seven amino acids (Glu-Leu-Leu-Val-Asp-Leu-Leu) with a lactone linkage.

• Journal of Microbiology and Biotechnology
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• 제16권5호
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• pp.716-723
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• 2006

A biosurfactant-producing bacterial strain was selected from oil-contaminated soil because of its ability to degrade crude oil and tributyrin $(C_{4:0})$. The strain was identified as Bacillus subtilis A8-8 based on its morphological, biochemical, and physiological characteristics. When B. subtilis A8-8 was grown with crude oil as the sole carbon source, the biosurfactant from the strain emulsified crude oil, vegetable oil, and hydrocarbons. Soybean oil was the optimum substrate for the emulsifying activity and emulsion stability of the biosurfactant, both of which were superior to those of several commercially available surfactants. The biosurfactant was purified by a procedure including HCl precipitation, methanol treatment, and silica-gel chromatography. The partially purified biosurfactant was analyzed by TLC (thin-layer chromatography), SDS-PAGE, and HPLC and it reduced the surface tension of water from 72 mN/m to 26 mN/m at a concentration of 30 mg/l. Therefore, the purified lipopeptide biosurfactant has strong properties as an emulsifying agent and acts as an emulsion-stabilizing agent.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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• pp.528-531
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• 2000

식물의 토양전염병을 유발하는 Pythium sp. 및 Rhizoctonia sp.에 대한 길항력과 생장속도를 고려한 RPI (relative performance indices) 기법을 이용하여 100 여개 후보균 중 #19, #42 등 길항균을 선정하였다. 후보균들은 일반적으로 Pythium sp.보다 Rhizoctonia sp.에 대한 길항력이 우수하였고 Bacillus sp.의 배양 상등액은 항진균활성과 blood agar에서 용혈현상을 보여 상등액으로부터 항균물질인 lipopeptide 계열물질을 분리하였다.

• KSBB Journal
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• 제14권6호
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• pp.737-741
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• 1999

본 연구에서 분리된 phenanthrene 분해균주의 대부분이 소수성을 나타내었고, 미생물계면활성제를 생산하면서 분해하는 균주보다 phenanthrene 분해능이 높았다. 균주 H6의 경우 phenanthrene 분해시 생산되는 미생물계면활성제는 포도당을 분해할 때도 생산되면 미생물의 성장과 더불어 증가하다가 stationary phase에서 일정하게 유지되었다. H6는 Bacillus subtilis로서 분리된 미생물계면활성제는 lipopeptide이었으며 H6에서 추출한 미생물계면활성제는 pH 2 이하에서 거의 회수되었고, 분산력이 Tween 80이나 Brij 30 등 화학계면활성제보다 높았고 열에 강한 특성을 보였다.

• 미생물학회지
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• 제54권3호
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• pp.244-253
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• 2018

이 연구는 분리된 토양 세균에 의해 생성된 항균물질의 효과를 평가하기 위해 수행되었다. 2000여개의 세균 분리주 중 Paenibacillus elgii DS381이 여러 인간 피부 상재균과 병원성 세균에 대해 높은 항균활성을 나타내었다. DS381 균주는 agar well diffusion test에서 모든 대상 세균과 효모에 대해 15.3~26.0 mm 직경의 저해대를 형성하였다. DS381이 생성한 항균 펩티드는 모든 대상 미생물에 낮은 최소저해농도 (0.039-5.000 mg/ml)를 나타내었다. DS381 균주는 lipopeptide 같은 생물계면활성제 생산을 나타내었는데, 배양 상등액의 표면장력을 60.0에서 40.3 mN/m으로 낮추었다. DS381은 또한 $1.56{\pm}0.13U/ml$의 chitinase 활성도 나타내었다. 이 결과들은 P. elgii DS381이 일부 중요한 인간 피부 상재균과 병원성 세균에 대한 효율적인 생물제어제로 사용될 수 있음을 가리킨다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XII)
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• pp.40-44
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• 2003

Soil-borne infectious disease including Pythium aphanidermatum and Rhizoctonia solani causes severe damage to plants, such as cucumber. This soil-borne infectious disease was not controlled effectively by chemical pesticide. Since these diseases spread through the soil, chemical agents are usually ineffective. Instead, biological control, including antagonistic microbe can be used as a preferred control method. An efficient method was developed to select an antagonistic strain to be used as a biological control agent strain. In this new method, surface tension reduction potential of an isolate was included in the ‘decision factor’ in addition to the other factors, such as growth rate, and pathogen inhibition rate. Considering these 3 decision factors by a statistical method, an isolate from soil was selected and was identified as Bacillus sp. GB16. In the pot test, this strain showed the best performance among the isolated strains. The lowest disease incidence rate and fastest seed growth was observed when Bacillus sp. GB16 was used. Therefore this strain was considered as plant growth promoting rhizobacteria (PGPR). The action of surface tension reducing component was deduced as the enhancement of wetting, spreading, and residing of antagonistic strain in the rhizosphere. This result showed that new selection method was significantly effective in selecting the best antagonistic strain for biological control of soil-borne infectious plant pathogen. The antifungal substances against P. aphanidermatum and R. solani were partially purified from the culture filtrates of Bacillus sp. GB16. In this study, lipopeptide possessing antifungal activity was isolated from Bacillus sp. GB16 cultures by various purification procedures and was identified as a surfactin-like lipopeptide based on the Fourier transform infrared spectroscopy (FT-IR), nuclear magnetic resonance (NMR), high performance liquid chromatography mass spectroscopy (HPLC-MS), and quadrupole time-of-flight (Q-TOF) ESI-MS/MS data. The lipopeptide, named GB16-BS, completely inhibited the growth of Pythium aphanidermatum, Rhizoctonia solani, Penicillium sp., and Botrytis cineria at concentrations of 10 and 50 mg/L, respectively. A novel method to prevent the foaming and to provide oxygen was developed. During the production of surface active agent, such as lipopeptide (surfactin), large amount of foam was produced by aeration. This resulted in the carryover of cells to the outside of the fermentor, which leads to the significant loss of cells. Instead of using cell-toxic antifoaming agents, low amount of hydrogen peroxide was added. Catalase produced by cells converted hydrogen peroxide into oxygen and water. Also addition of corn oil as an oxygen vector as well as antifoaming agent was attempted. In addition, Ca-stearate, a metal soap, was added to enhance the antifoam activity of com oil. These methods could prevent the foaming significantly and maintained high dissolved oxygen in spite of lower aeration and agitation. Using these methods, high cell density, could be achieved with increased lipopeptide productivity. In conclusion to produce an effective biological control agent for soil-borne infectious disease, following strategies were attempted i) effective screening of antagonist by including surface tension as an important decision factor ii) identification of antifungal compound produced from the isolated strain iii) novel oxygenation by $H_2O_2-catalase$ with vegetable oil for antifungal lipopeptide production.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XII)
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• pp.240-244
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• 2003

A novel integrated method for increasing dissolved oxygen concentration in culture media has been developed. It involves adding hydrogen peroxide to the medium, which is then decomposed to oxygen and water by catalase and adding vegetable oil to the medium as antifoam agent and oxygen vector. A new apparatus for automated addition of hydrogen peroxide to the bioreactor to keep the dissolved oxygen concentration constant over the range $10-100%\;{\pm}\;5%$ was tested. A significant increase (over threefold) of cultivation time was obtained while the dissolved oxygen concentration remained stable ($30%\;{\pm}\;5%$). Therefore, use of corn oil mixed with Ca-stearate as oxygen vector and antifoam and hydrogen peroxide as oxygen source to control excessive foam that was generated by microorganism biosurfactant, GB16-BS produced at Bacillus sp. GB16 cultivation was appropriate for stable cultivation.