• Journal of Microbiology and Biotechnology
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• v.18 no.1
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• pp.63-66
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• 2008

Anthracene is a PAH that is not readily degraded, plus its degradation mechanism is still not clear. Thus, two strains of anthracene-degrading bacteria were isolated from long-term petroleum-polluted soil and identified as Sphingomonas sp. 12A and Pseudomonas sp. 12B by a 16S rRNA sequence analysis. To further enhance the anthracene-degrading ability of the two strains, the biosurfactants produced by Pseudomonas aeruginosa $W_3$ were used, which were characterized as rhamnolipids. It was found that these rhamnolipids dramatically increased the solubility of anthracene, and a reverse-phase HPLC assay showed that the anthracene degradation percentage after 18 days with Pseudomonas sp. 12B was significantly enhanced from 34% to 52%. Interestingly, their effect on the degradation by Sphingomonas sp. 12A was much less, from 35% to 39%. Further study revealed that Sphingomonas sp. 12A also degraded the rhamnolipids, which may have hampered the effect of the rhamnolipids on the anthracene degradation.

• Journal of the Korean Applied Science and Technology
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• v.38 no.5
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• pp.1219-1228
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• 2021

Rhamnolipids are recognized as eco-friendly biosurfactants and produced by the bio-process employing bacteria. Pseudomons aeruginosa is well-known to produce rhamnolipids in high yield during fermentation process. Rhamnose and 𝛽-hydroxylated fatty acid are main chemicals for rhamnolipids, which are produced in the form of congener mixtures. In this paper, the synthetic mechanism of rhamnolipids within bacteria cells was presented in part and foam control technologies were qualitatively described. Foam control during fermentation process was important to regulate a continuous process. During last decade, the technologies are developed enough to challenge to a commercial-scale production. In particular, rhamnolipids will be more valuable if these can be applicable to value-added chemicals, such as medicines.

• Environmental Engineering Research
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• v.13 no.1
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• pp.8-13
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• 2008

The extent of solubility enhancement by biosurfactant was examined at various pHs prior to the biodegradation experiments. The molar solubilization ratio (MSR) was calculated from the batch solubilization experiments and the highest MSR was detected at pH 5. The effect of the biosurfactant, rhamnolipids, on the phenanthrene mineralization in soil-water system was investigated. The strain 3Y was selected for the mineralization assay and large amounts of phenanthrene were degraded at neutral pH in soil-water system without the biosurfactant. The addition of 150 mg/L rhamnolipids showed no effect on mineralization of phenanthrene in soil-water system, and total mineralization rates after 6 weeks incubation at each pH showed no differences in presence and absence of rhamnolipids. Our result indicated that the toxic effect of rhamnolipids can disappear when soil particles exist, and also the enhanced solubility of phenanthrene does not work for mineralization enhancement in this soil-water system.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.329-331
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• 2000

A marine bacterium, Pseudomonas aeruginosa BYK-2 KCTC 18012P was immobilized in modified polyvinyl alcohol for the continuous production of rhamnolipids. The stability of rhamnolipids production, the mechanical strength of beads and the scanning electron microscope of immobilized cell were determined in a repeated batch culture. The rhamnolipids production was maintained $80{\sim}90%$ stability of initial production, and the mechanical strength also was stable during the repeated batch culture more than 14 cycles. In the case of SEM studies, the internal distribution pattern of the cell entrapped in modified PVA beads was observed. On the basis of optimal conditions, the continuous culture was investigated in 1.8L air lift bioreactor. The result suggested 0.1g/h rhamnolipids was obtained from 1%(v/v) fish oil continuously in conditions of 1.2L working volume, 0.5vvm and 20ml/h flow rate.

• 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.22 no.1
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• pp.69-79
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• 2012

In an ongoing survey of the bioactive potential of microorganisms from Ladakh, India, the culture medium of a bacterial strain of a new Pseudomonas sp., strain ICTB-745, isolated from an alkaline soil sample collected from Leh, Ladakh, India, was found to contain metabolites that exhibited broad-spectrum antimicrobial and biosurfactant activities. Bioactivity-guided purification resulted in the isolation of four bioactive compounds. Their chemical structures were elucidated by $^1H$ and $^{13}C$ NMR, 2D-NMR (HMBC, HSQC, $^1H$,$^1H$-COSY, and DEPT-135), FT-IR, and mass spectroscopic methods, and were identified as 1-hydroxyphenazine, phenazine-1-carboxylic acid (PCA), rhamnolipid-1 (RL-1), and rhamnolipid-2 (RL-2). These metabolites exhibited various biological activities like antimicrobial and efficient cytotoxic potencies against different human tumor cell lines such as HeLa, HepG2, A549, and MDA MB 231. RL-1 and RL-2 exhibited a dose-dependent antifeedant activity against Spodoptera litura, producing about 82.06% and 73.66% antifeedant activity, whereas PCA showed a moderate antifeedant activity (63.67%) at 60 ${\mu}g/cm^2$ area of castor leaf. Furthermore, PCA, RL-1, and RL-2 exhibited about 65%, 52%, and 47% mortality, respectively, against Rhyzopertha dominica at 20 ${\mu}g/ml$. This is the first report of rhamnolipids as antifeedant metabolites against Spodoptera litura and as insecticidal metabolites against Rhyzopertha dominica. The metabolites from Pseudomonas sp. strain ICTB-745 have interesting potential for use as a biopesticide in pest control programs.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.212-215
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• 2000

Recombinant bioluminescent bacterial strains that use specific promoters fused to the bioluminescence genes (lux genes) have been applied in environmental monitoring. Advantages of using recombinant bioluminescent bacteria as blosensing cells include rapid responses, low costs, and improved reproducibility. In this study, a recombinant Escherichia coli, GC2, containing a lac::luxCDABE fusion immobilized with solid agar media and glass beads was used to estimate the effect of this soil flushing technique. This bacterium constitutively emits light under normal conditions (no toxic chemicals). When growth and metabolism of these bioluminescent bacteria is inhibited by their exposure to toxic chemicals, the bioluminescence (BL) is reduced. A biosurfactant, rhamnolipids, was used to extract phenanthrene from the soil after flushing.

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.434-437
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• 2003

A new bacterial strain, was isolated from activated sludge, identified and named P. aeruginosa EMS1. The new strain produced surface-active rhamnolipids by batch cultivation in mineral salts medium with waste flying oils. The mutant strain KH7, designated P. aeruginosa EMS1, derived by random mutagenesis with N-methyl-N-nitro-N-nitrosogoanidine treatment producing high levels of the biosurfactants was selected by an ion-pair plate assay. The mutant strain KH7 showed 4-5 times more hydrocarbon emulsification as compared to the parent when grown on waste frying oils and various hydrocarbons. Furthermore, P. aeruginosa EMS1 and mutant strain KH7 was also able to use whey as a co-substrate for growth and biosurfactant production. As results of this study, mutant strain KH7 is a very efficient biosurfactant producer, and its culture conditions are relatively inexpensive and economical. Rhamnolipid is synthesized by the rhlAB-encoded rhamnosyltransferase. To be convinced of these genes, we performed PCR based on P. aeruginosa PAO1 whole-genome database. rhl gene cluster nucleotide and amino acid sequences were compared for both parent and mutant. Comparison of nucleotide sequence of rhlAB, there were usually terminal's codons exchange.

• Biotechnology and Bioprocess Engineering:BBE
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• v.3 no.2
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• pp.61-66
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• 1998

Glycolipids produced by Pseudomonas aeruginosa YPJ-80 were characterized by chromatographic and spectorscopic techniques as a mixture of two rhamnolipids. For recovery of glycolipids from the culture broth, various isolation methods including ultrafiltration, adsorption and solvent extraction were compared. Ultrafiltration showed the best results in terms of glycolipids recovery. Further purification for spectroscopic analysis was carried out by adsorption chromatography and preparative thin layer chromatography. From the spectroscopic analysis, such as IR spectroscopy. FAB-MS, 1H-NMR and 13C-NMR and hydrolysis analysis, the glycolipids were identified as L-${\alpha}$-rhamnopyranosyl-${\beta}$-hydroxydecanoly-${\beta}$-hydroxydecanoate and 2-O-${\alpha}$-L-rhamnopyranosyl-${\alpha}$-L-rhamnopyranosyl-${\beta}$-hydroxydecanoyl-${\beta}$-hydroxydecanoate. Monorhamnolipid and dirhamnolipid lowered the surface tension of water to 28.1 mN/M and 29.3 mN/m, respectively.

• Natural Product Sciences
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• v.25 no.2
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• pp.172-180
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• 2019

Infections with Pseudomonas aeruginosa are difficult to treat not only because it is often associated with multidrug-resistant infections but also it is able to form biofilm. The aim of this study was to evaluate the antibiofilm and anti-Quorum Sensing (QS) activities of Thymbra capitata essential oils (EOs) against Beta Lactamase (BL) producing P. aeruginosa and the reference strain P. aeruginosa 10145. GC/MS analysis showed that thymol (23.25%) is the most dominant compound in T. capitata EOs. The MICs of T. capitata EOs against P. aeruginosa (BL) and P. aeruginosa 10145 were 1.11%. At sub MIC (0.041, 0.014 and 0.0046%), the EOs of T. capitata remarkably inhibited the biofilm formation of both strains tested and complete inhibition of the biofilm formation was reported at 0.041%. The EOs of T. capitata were found to inhibit the swarming motility, aggregation ability and hydrophobic ability of P. aeruginosa (BL) and P. aeruginosa 10145. Interestingly, the EOs of T. capitata reduce the production of three secreted virulence factors that regulated by QS system including pyocyanin, rhamnolipids and LasA protease. The potent antibiofilm and anti-QS activities of T. capitata EOs can propose it as a new antibacterial agent to control pseudomonas infections.