• 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.

• 한국미생물·생명공학회지
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• 제33권4호
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• pp.295-301
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• 2005

한국 전통 젓갈로부터 항균성 lipopeptide를 생산하는 JKK238 균주를 분리하여 16S rDNA및 세포벽의 지방산 조성 분석 등에 따라 Bacillus subtilis로 동정하였다. JKK238 균주를 3일간 배양한 TSB 액체배지의 $25\%$ ammonium sulfate 침전물이 대부분의 lipopeptide를 포함하고 이들이 다양한 식물병원성 곰팡이나 세균의 성장을 강력히 저해함을 알 수 있었다. 또한 FPLC에 의해 분리 및 정제된 각 lipopeptide의 Maldi-tof분석에서, 이 균주가 iturin A, fengycin, surfactin과 같은 세 종류의 lipopeptide의 isomer들을 동시에 생산하는 균주임을 밝혔다.

• Journal of Microbiology and Biotechnology
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• 제28권5호
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• pp.784-795
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• 2018

Bacillus amyloliquefaciens Pc3 was isolated from Antarctic seawater with antifungal activity. In order to investigate the metabolic regulation mechanism in the biosynthesis of lipopeptides in B. amyloliquefaciens Pc3, GC/MS-based metabolomics was used when exogenous indole was added. The intracellular metabolite profiles showed decreased asparagine, aspartic acid, glutamine, glutamic acid, threonine, valine, isoleucine, hexadecanoic acid, and octadecanoic acid in the indole-treated groups, which were involved in the biosynthesis of lipopeptides. B. amyloliquefaciens Pc3 exhibited a growth promotion, bacterial total protein increase, and lipopeptide biosynthesis inhibition upon the addition of indole. Besides this, real-time PCR analysis further revealed that the transcription of lipopeptide biosynthesis genes ituD, fenA, and srfA-A were downregulated by indole with 22.4-, 21.98-, and 26.0-fold, respectively. It therefore was speculated that as the metabolic flux of most of the amino acids and fatty acids were transferred to the synthesis of proteins and biomass, lipopeptide biosynthesis was weakened owing to the lack of precursor amino acids and fatty acids.

• 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.

• 미생물학회지
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• 제55권3호
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• pp.286-288
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• 2019

벼 종자에서 분리한 Pseudomonas parafulva PpaJBCS1880 (PpaJBCS1880) 균주는 lipopeptide를 분비하여 식물의 세균 병원균에 대해 강력한 항균력을 나타냈다. 또한, PpaJBCS1880는 콩불마름병의 발생을 억제하였을 뿐만 아니라, 벼의 생육을 촉진하였다. 이에 따라, 본 연구에서는 PpaJBCS1880 균주의 전체염기서열을 해독하고 분석하였는데, 총 염기서열은 5,208,480 bp였고, GC 함량은 63.4%였다. 염색체는 4,487개의 단백질을 암호화하였고, 19개의 rRNA와 74개의 tRNA로 구성되어 있었다. 유전체의 분석을 통해 2차 대사산물인 lipopeptide, pyoverdine, phenazine 및 hydrogen cyanide 등을 생산하는 것을 확인하였는데, 이들 대사산물에 의해 항균력, 생물방제 및 생육촉진 효과를 나타내는 것으로 판단된다.

• 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.

• Bulletin of the Korean Chemical Society
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• 제31권1호
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• pp.205-208
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• 2010

• 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.

• The Plant Pathology Journal
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• 제36권5호
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• pp.491-496
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• 2020

An understanding of the contribution of secondary metabolites (SMs) to the antagonistic and biocontrol activities of bacterial biocontrol agents serves to improve biocontrol potential of the strain. In this study, to evaluate the contribution of each SM produced by Pseudomonas fluorescens NBC275 (Pf275) to its antifungal and biocontrol activity, we combined in silico analysis of the genome with our previous study of transposon (Tn) mutants. Thirteen Tn mutants, which belonged to 6 biosynthetic gene clusters (BGCs) of a total 14 BGCs predicted by the antiSMASH tool were identified by the reduction of antifungal activity. The biocontrol performance of Pf275 was significantly dependent on 2,4-diacetylphloroglucinol and pyoverdine. The clusters that encode for arylpolyene and an unidentified small linear lipopeptide influenced antifungal and biocontrol activities. To our knowledge, our study identified the contribution of SMs, such as a small linear lipopeptide and arylpolyene, to biocontrol efficacy for the first time.