• 한국생물공학회:학술대회논문집
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• 2003.04a
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• pp.393-397
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• 2003

P. aeruginosa F722 produces biosurfactant (BS) while degrading hydrocarbons. BS production was 0.78 $g/{\ell}$ on the C-medium. However, BS production increased by 1.66 $g/{\ell}$ on the condition of 0.05% (w/v) $NH_4Cl+0.1%$ (w/v) yeast extract and 3.0% (w/v) glucose, which was proved to be advantageous to BS production. In the condition of aeration of 1.0 liter per minute (LPM), BS production was increased 20% (1.94 $g/{\ell}$)more than 1.66 $g/{\ell}$ produced when the air was not supplied. Moreover, the velocity of glucose degradation at both of log and stationary growth phases increased from 0.25 and 0.18 $h^{-1}$ to 0.33 and 0.29 $h^{-1}$ respectively when the air was supplied. Besides, BS activity was more stabilized on the condition of air supply.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.6
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• pp.471-476
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• 2006

Pseudomonas aeruginosa F722 produces a biosurfactant (BS) during its degradation of carbon and hydrocarbon compounds. The culture conditions for upgrading the biosurfactant productivity were investigated. The concentration of the biosurfactant produced by P. aeruginosa F722 was 0.78 g/L in C-medium; however, this increased to 1.66 g/L in BS medium, which was experimentally adjusted to optimal conditions. $NaNO_{2}$ was found to be most effective for microbial growth, with an $O.D_{600nm}$ of 1.18 for 0.1 % $NaNO_{2}$. Microbial growths, according to the $O.D_{600nm}$ were 2.53, 2.68, 2.89, and 2.87 for glucose, glycerol, $n-C_{10},\;and\;n-C_{22}$, respectively. Clear zone diameters (cm), indicating biosurfactant activity, were 9.0, 8.8, 5.7, and 8.5 for glucose, glycerol, $n-C_{10},\;and\;n-C_{22}$, respectively. Microbial growth was not consistent with the biosurfactant activity. The best biosurfactant activity was found with a C/N ratio of 20. Under optimal culture condition, the average surface tension decreased from 70 to 30 mN/m after 5 days. With aeration of 1.0 vvm, the biosurfactant produced increased to 1.94 g/L (up to 20%) compared to that of 1.66 g/L with no aeration. With aeration, the velocities of glucose degradation during both the log and stationary growth phases increased from 0.25 and $0.18\;h^{-1}$ to 0.33 and $0.29\;h^{-1}$, respectively, and the time for the culture to arrive at the maximum clear zone diameter became shorter, from 80 down to 60 h with no aeration.

• Journal of Microbiology and Biotechnology
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• v.7 no.3
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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.

• KSBB Journal
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• v.18 no.2
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• pp.145-148
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• 2003

The hydrogen-producting bacterium was isolated from fresh water and identified as Enterbacter cloacae. The isolated was named Enterobacter cloacae YJ-1. In batch culture, The optimum cultivation temperature and pH of strain YJ-1 was 35$^{\circ}C$ and 7.5, respectively. All of the added glucose was consumed completely during fermentation even though pH was not controlled. Amount of hydrogen produced on each condition of 2% glucose, 4% sucrose and 5% fructose was 950, 1000 and 948 mL/L, respectively and resulted in increasing hydrogen production approximately 2.5-times more than controlled condition. The macimum hydrogen production was obtained when 50mM phosphate was added. was obtained when 50mM phosphate was added. In repeated0batch culture, yeast extract, but the production amount was not changed on the condition of over 0.5%, Most of the organic acides produced during the fermentation were formic and acetic acid, and propionic acid was moiety also generated.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.626-629
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• 2000

The Pseudomonas aeruginosa BYK-2(KCTC 18012p) produced three kinds of glycolipids on olive oil as a substrate and purified two types of major glycolipids(Rf=0.48, BS-1; Rf=0.65, BS-2) using silica gel chromatography, TLC, HPLC, etc. From the analysis of the chemical structure, the glycolipid of BS-1 was estimated as rhamnolipid($2-O-{\alpha}-L-rhamnopyranosyl- {\alpha}-L-rhamnopyranosyl-{\beta}-hydroxyldecanoyl-{\beta}-hydroxydecanoic$ acid; M.W. 650) and BS-2 was detected as rhamnolipid methyl ester($2-O-{\alpha}-L-rhamnopyranosyl-{\alpha}-L-rhamnopyranosyl-{\beta}-hydroxyldecanoyl-{\beta}-hydroxydecanoic$ acid methyl ester; M.W. 664) by FT-IR, FAB Mass spectrometry, $^1H-NMR$, $^{13}C$ FT-NMR, DEPT, 2D-NMR (TOCSY, RELAY, NOESY, HSQC, HMBC). In particular, It was found that a marine bacterium Pseudomonas aeruginosa BYK-2(KCTC 18012P) remarkably produced rhamnolipid and rhamnolipid methyl ester simultaneously.

• Journal of Microbiology and Biotechnology
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• v.20 no.7
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• pp.1061-1068
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• 2010

The biological synthesis of nanoparticles has gained considerable attention in view of their excellent biocompatibility and low toxicity. We isolated and purified rhamnolipids from Pseudomonas aeruginosa strain BS-161R, and these purified rhamnolipids were used to synthesize silver nanoparticles. The purified rhamnolipids were further characterized and the structure was elucidated based on one- and two-dimensional $^1H$ and $^{13}C$ NMR, FT-IR, and HR-MS spectral data. Purified rhamnolipids in a pseudoternary system of n-heptane and water system along with n-butanol as a cosurfactant were added to the aqueous solutions of silver nitrate and sodium borohydride to form reverse micelles. When these micelles were mixed, they resulted in the rapid formation of silver nanoparticles. The synthesized nanoparticles were characterized by UV-Visible spectroscopy, transmission electron microscopy, and energy dispersive X-ray spectroscopy (EDS). The nanoparticles formed had a sharp adsorption peak at 410 nm, which is characteristic of surface plasmon resonance of the silver nanoparticles. The nanoparticles were monodispersed, with an average particle size of 15.1 nm (${\sigma}={\pm}5.82$ nm), and spherical in shape. The EDS analysis revealed the presence of elemental silver signal in the synthesized nanoparticles. The formed silver nanoparticles exhibited good antibiotic activity against both Grampositive and Gram-negative pathogens and Candida albicans, suggesting their broad-spectrum antimicrobial activity.

• Journal of Microbiology and Biotechnology
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• v.17 no.1
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• pp.67-73
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• 2007

The present study compared the microbial diversity and activity during the application of various bioremediation processes to crude oil-contaminated soil. Five different treatments, including natural attenuation (NA), biostimulation (BS), biosurfactant addition (BE), bioaugmentation (BA), and a combined treatment (CT) of biostimulation, biosurfactant addition, and bioaugmentation, were used to analyze the degradation rate and microbial communities. After 120 days, the level of remaining hydrocarbons after all the treatments was similar, however, the highest rate (k) of total petroleum hydrocarbon (TPH) degradation was observed with the CT treatment (P<0.05). The total bacterial counts increased during the first 2 weeks with all the treatments, and then remained stable. The bacterial communities and alkane monooxygenase gene fragment, alkB, were compared by denaturing gradient gel electrophoresis (DGGE). The DGGE analyses of the BA and CT treatments, which included Nocardia sp. H17-1, revealed a simple dominant population structure, compared with the other treatments. The Shannon-Weaver diversity index (H') and Simpson dominance index (D), calculated from the DGGE profiles using 16S rDNA, showed considerable qualitative differences in the community structure before and after the bioremediation treatment as well as between treatment conditions.

• 한국생물공학회:학술대회논문집
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• 2003.04a
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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.

• 한국생물공학회:학술대회논문집
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• 2003.04a
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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.