ABSTRACT
Discovering new antibacterial agents is crucial to addressing the increasing risk of bacterial infections induced by antimicrobial resistance in food and agricultural industries. Here, biocompatible acidic-type sophorolipids (ASLs) and glucolipids (GLs) prepared via chemical modification of natural sophorolipids from fermentation were functionalized via amide modification for use as potential antibacterial agents. It was found that the arginine methyl ester derivative of GLs (GLs-d-Arg-OMe) showed excellent antibacterial activity, killing more than 99.99% of Escherichia coli at 200 mg/L. The sterilization dosage of the GLs against Bacillus subtilis, Bacillus cereus, and Staphylococcus aureus was 16-64 mg/L, in contrast to 32-64 mg/L for the fungus Candida albicans. In particular, GLs-d-Arg-OMe showed the best biocompatibility with a therapeutic index of up to 18. It was shown that amide modification of glycolipids can effectively improve antibacterial activity while maintaining biocompatibility, which can be exploited for the development of novel antibiotics in food and agricultural fields.
Subject(s)
Anti-Bacterial Agents , Glycolipids , Anti-Bacterial Agents/pharmacology , Glycolipids/chemistry , Candida albicans , Bacillus subtilis , Bacillus cereus , Microbial Sensitivity TestsABSTRACT
This investigation developed a novel strategy for efficiently preparing glucolipids (GLs) by chemically modifying crude sophorolipids. Running this strategy, crude sophorolipids were effectively transformed into GLs through deglycosylation and de-esterification, with a yield of 54.1%. The acquired GLs were then purified via stepwise extractions, and 66.2% of GLs with 95% purity was recovered. GLs are more hydrophobic and present a stronger surface activity than acidic sophorolipids (ASLs). More importantly, these GLs displayed a superior antimicrobial activity to that of ASLs against the tested Gram-positive food pathogens, with a minimum inhibitory concentration of 32-64 mg/L, except against E. coli . This activity of GLs is pH-dependent and especially more powerful under acidic conditions. The mechanism involved is possibly associated with the more efficient adsorption of GLs, as demonstrated by the hydrophobicity of the cell membrane. These GLs could be used as antimicrobial agents for food preservation and health in the food industry.
Subject(s)
Escherichia coli , Surface-Active Agents , Surface-Active Agents/chemistry , Escherichia coli/metabolism , Oleic Acids/chemistry , Food Industry , Glycolipids/chemistryABSTRACT
Recent upgrades in the electrochemiluminescence (ECL) technique showcased its brilliant knack in probing microscopic biointerfacial events, many of which were actually underlain by the ionotropic membrane processes, yet not being ostensive. Here, by modeling an artificial lipoid-supported porin ensemble, we explore and establish the ECL potency in profiling ion-channel activities. A lipophilic hollowed construct dubbed ZnPC was made out of the dynamic covalent chemistry, and its unique geometry was characterized that configured stoichiometric ECL-emissive units in a cubic stance; while the aliphatic vertices of ZnPC helped it safely snorkel and steadily irradiate in a biofilm fusion. After expounding basic ECL properties, the brightness was traced out in response to halogen contents that was lit up by F-/Cl- but down by Br-/I-. The overall pattern fitted the Langmuir isotherm, from which the membrane-binding strengths of the four were analyzed, compared, and collaterally examined in impedimetrics. On the other hand, one could derive anionic transmembrane kinetics from the time-dependent ECL statistics that pinpointed the ECL signaling via the nanocage-directed mass-transfer pathway. More data mining unveiled an ECL-featured Hofmeister series and the thermodynamic governing force behind all scenes. Finally, combining with halide-selective fluorometry, the synthetic conduit was identified as an ECL symporter. In short, this work develops a novel ECL model for the evaluation of life-mimicking membrane permeation. It might intrigue the outreach of ECL applications in the measurement of diverse surface-confined transient scenarios, e.g., in vitro gated ion or molecule trafficking, which used to be handled by nanopore and electrofluorochromic assays.
Subject(s)
Electrochemical Techniques , Luminescent Measurements , Electrochemical Techniques/methods , Luminescent Measurements/methods , PhotometryABSTRACT
The development of new preservatives is an ongoing investigation in the food industry, especially those which are safe and environmentally friendly. In this study, biosurfactant sophorolipids (SLs) functionalized with amino acids were developed as efficient preservative agents. SLs were first isolated from fermentation broth by Candida bombicola ATCC 22214, hydrolyzed, and purified by extraction. The typical recovery is around 70%, while the extracted material consists of over 90% deacetylated acidic SLs (SL-COOH). Four types of SL derivatives were then synthesized via dicyclohexylcarbodiimide amidation reactions from prepared SL-COOH. Among the derivatives produced, the arginine SL conjugates (SL-d-Arg) displayed the highest activity against Gram-positive bacteria and fungi and even inhibited the cell growth of Gram-negative bacteria and mildew. Furthermore, the arginine conjugates performed the broadest antimicrobial activity among the derivatives evaluated. The sterilization dosage of the arginine conjugates against the food-spoilage pathogen Bacillus spp. was 63-125 mg/L, in contrast to 250 mg/L for the enterotoxin producer Staphylococcus aureus and 500 mg/L for fungi. More importantly, SL-d-Arg displayed excellent biocompatibility, with a therapeutic index of over 7.94. SL-d-Arg has excellent potential as an alternative to traditional chemical preservatives.
Subject(s)
Amino Acids , Glycolipids , Oleic Acids , SaccharomycetalesABSTRACT
Rhamnolipids have extensive potential applications and are the most promising biosurfactants for commercialization. The efficient and accurate identification and analysis of these are important to their production, application and commercialization. Accordingly, significant efforts have been made to identify and analyse rhamnolipids during screening of producing strains, fermentation and application processes. Cationic cetyltrimethylammonium bromide-methylene blue (CTAB-MB) test combines a series of indirect assays to efficiently assist in the primary screening of rhamnolipids-producing strains, while the secretion of rhamnolipids by these strains can be identified through TLC, FTIR, NMR, electrospray ionization mass spectrometry (ESI-MS) and HPLC-MS analysis. Rhamnolipids can be quantified by colorimetric methods requiring the use of concentrated acid, and this approach has the advantages of reliability, simplicity, low-cost and excellent reproducibility with very low technological requirements. HPLC-MS can also be employed as required as a more accurate quantification method. In addition, HPLC-ELSD has been established as the internationally acceptable measure of rhamnolipids for commercial purposes. The preparation of well-accepted rhamnolipids standards and modifications of analysis operations are essential to further enhance the accuracy and improve the simplicity of rhamnolipid analysis.Key points⢠Current status of R&D works on determination of rhamnolipids is listed⢠Advantages and disadvantages of various types analysis are summarized⢠Limitations of current rhamnolipid quantification are discussed Graphical abstract.
Subject(s)
Glycolipids , Pseudomonas aeruginosa , Chromatography, High Pressure Liquid , Reproducibility of ResultsABSTRACT
Tissue and cellular stiffening is associated with pathologies including fibrosis and cancer. Healthy cells also exhibit a wide range of membrane cortical tensions, which have been studied in the field of mechanobiology. Here, we quantify the mechanosensitivity of the lysis agent the di-rhamnolipid (RHA), which is a bacterially produced biosurfactant. RHA exhibited selective lysis correlated strongly with cortical membrane tension in osteoblasts, smooth muscle cells, fibroblasts, epithelial cells, and erythrocytes. Reducing cortical membrane tension by cytoskeleton inhibitors (cytochalasin D and nocodazole) or osmotic regulators (sucrose, polyethylene glycol, and nonionic surfactants) attenuated the RHA toxicity. The selective toxicity of RHA toward human chronic myeloid leukemia K562 cells over healthy blood cells suggests a potential therapy for blood cancer. Targeted killing of myofibroblasts transformed from either fibroblasts or epithelial cells indicates its antifibrotic effect. Combined, these studies showed the potential for specific targeting of cells with differential mechanical properties rather than chemical or biological pathways.
Subject(s)
Pseudomonas aeruginosa , Surface-Active Agents , Glycolipids , Humans , Myofibroblasts , Surface-Active Agents/pharmacologyABSTRACT
The global market for rhamnolipids production holds great promise, and is in need of an economically viable mass-production scheme. Accordingly, several strategies have been employed to improve the efficiency of rhamnolipid production in the past few decades. Currently, rhamnolipids can be produced by Pseudomonas aeruginosa at a high yield (over 70 g/L) when vegetable oil is used as the carbon source under optimized fed-batch cultivations. However, severe foaming during rhamnolipid fermentation inhibits scaling-up and production efficiency. Stop valve was found to effective break the extremely stable rhamnolipids foams during fermentation, and production efficiency of rhamnolipids was highly improved, while its scale-up mechanism needs further study. In addition, the combination of both chemical and mechanical approaches is likely to be more efficiently resolving the foam problem existed in rhamnolipids fermentation than either chemical or mechanical methods alone.
Subject(s)
Glycolipids , Pseudomonas aeruginosa , Carbon , Fermentation , Surface-Active AgentsABSTRACT
Efficient oil separation is the most desirable, but still challenging solution for the waste crude oil problem. This study developed biosurfactant surfactin as a novel pH-switchable biodemulsifier for efficient oil separation. As found, surfactin demulsification achieved a quite well oil separation ratio of over 95% on model emulsions after 20â¯minâ¯at 50⯰C. The validity of this demulsification process should be mainly based on the readily lost stabilization ability of surfactin in emulsions triggered by acid addition. Then, surfactin (0.2â¯g/L) treatment with the aid of ethanol (2%) to improve its distribution could recover over 95% of oil from waste crude oil. After treated by surfactin, the separated oil phase contains tiny water (less than 0.5%) and thus can be reused for resource recycling to reach a compromised balance between satisfying the strict environmental regulations and decreasing the high treatment costs. Hence, in consideration of high demulsification efficiency, environmental-friendly properties and cost-efficiency, surfactin has a great potential for industrial applications for oil recovery from waste crude oil which is a severe problem presents in most of the petroleum-related factories.
Subject(s)
Emulsions/chemistry , Fuel Oils , Petroleum , Surface-Active Agents/pharmacology , Hydrogen-Ion ConcentrationABSTRACT
Parkinson disease-associated mutations within the GTPase domain Ras of complex proteins (ROC) of leucine rich repeat kinase 2 (LRRK2) result in an abnormal over-activation of its kinase domain. However, the mechanisms involved remain unclear. Recent study has shown that LRRK2 G-domain cycles between monomeric and dimeric conformations upon binding to GTP or guanosine diphosphate, and that the Parkinson's disease (PD)-associated R1441C/G/H mutations impair the G-domain monomer-dimer dynamics and trap the G-domain in a constitutive monomeric conformation. That led us to question whether other disease-associated mutations in G-domain would also affect its conformation. Here, we report that another PD-associated N1437H mutation also impairs its monomer-dimer conformational dynamics and GTPase activity. In contrast with mutations at R1441, ROCN1437H was found to be locked in a stable dimeric conformation in solution and its GTPase activity was â¼4-fold lower than that of the wild-type. Furthermore, the N1437H mutation reduced the GTP binding affinity by â¼2.5-fold when compared with other pathogenic G-domain mutations. Moreover, ROCN1437H was found to have a slower GTP dissociation rate, indicating that N1437H might interrupt the nucleotide exchange cycle. Taken together, our data support that conformational dynamics is important for LRRK2 GTPase activity and that the N1437H mutation impairs GTPase activity by locking the ROC domain in a persistently dimeric state.-Huang, X., Wu, C., Park, Y., Long, X., Hoang, Q. Q., Liao, J. The Parkinson's disease-associated mutation N1437H impairs conformational dynamics in the G domain of LRRK2.
Subject(s)
Leucine-Rich Repeat Serine-Threonine Protein Kinase-2/chemistry , Leucine-Rich Repeat Serine-Threonine Protein Kinase-2/metabolism , Parkinson Disease/genetics , Blotting, Western , Chromatography, Gel , Circular Dichroism , Electrophoresis, Polyacrylamide Gel , GTP Phosphohydrolases/chemistry , GTP Phosphohydrolases/genetics , GTP Phosphohydrolases/metabolism , Humans , Leucine-Rich Repeat Serine-Threonine Protein Kinase-2/genetics , Mutation/genetics , Protein ConformationABSTRACT
It is of great practical importance but rarely reported to design a multifunctional scavenger for water purification. In this study, we describe a sophisticated preparation of an inorganic/organic composite sample for the simultaneous removal of anionic dyes and metal ions, as well as disinfection. The sample has a stable structure formed by the covalent connection between a magnetic silica (MS) core and a polyethylenimine derived quaternary ammonium compound (QAC) corona. We characterized the sample in details by SEM, TEM, EDX, FT-IR, XRD, TGA, VSM, and zeta potential. Our QAC-MS sample exhibited superior performance and reusability in the disinfection and adsorption experiments towards acid fuchsin and Cu2+. With the virtue of easy separation from solution, our sample should be an ideal candidate for water purification application.
ABSTRACT
In the present study, the pH-regulated emulsification activity of surfactin was studied and its potential application in oil separation towards enhanced oil recovery (EOR) was investigated. As demonstrated, surfactin can stabilize emulsions quite well beyond pH 7.4. An oil emulsification ratio of about 98% was obtained at pH 11.0; while this emulsification activity was rapidly and completely lost when pH decreased to below 3.0, having an oil separation ratio of over 98%. This pH-sensitive property is probably due to surfactin dissolution-precipitation induced by the ionization-protonation of a carboxyl group in its structure under alkaline or acidic conditions. This property allows oil emulsification or oil separation to be readily achieved via simple pH adjustments when surfactin is used as an emulsifier. Furthermore, surfactin sustained its activity after demulsification and can be readily reused many times. The above obtained results indicated surfactin-based EOR processes have great application feasibility.
Subject(s)
Refuse Disposal , Emulsifying Agents , Emulsions , Hydrogen-Ion Concentration , Surface-Active AgentsABSTRACT
Rhamnolipids are the most promising biosurfactants, have widespread applications in many fields. However, low yield and productivity in fermentation caused a high production cost and thus prohibited the bulk applications of rhamnolipids in industry. In this study, a sequential fed-batch fermentation process with fill-and-draw operation was developed to improve rhamnolipids production. By utilizing this strategy, the total produced rhamnolipids reached over 150g/L, had a 163% and 102% increase over the traditional batch and fed-batch processes, respectively. This remarkable high production efficiency was achieved by the well-maintained high productivity of 0.4g/Lh for a period of 17 d. Astonishingly, the conversion yield was high as 84%, while this value was only 53.2% and 42.7% in the traditional batch and fed-batch process, respectively. The high-efficiency rhamnolipids production in this sequential fed-batch fermentation could be largely explained by a high presence of cell coupled with the replenishment of nutrients and dilution of toxic byproducts via fill-and-draw operation. In all, this validated fermentation strategy offers a great prospect for high-efficiency production of rhamnolipids in industry.
Subject(s)
Bioreactors , Glycolipids/chemistry , Batch Cell Culture Techniques , Surface-Active Agents/chemistryABSTRACT
In this study, a stop valve was used as a foam breaker for dealing with the massive overflowing foam in rhamnolipid fermentation. As found, a stop valve at its tiny opening could break over 90% of the extremely stable rhamnolipid foam into enriched liquid when foam flows through the sharp gap in valve. The efficient foam-control by the stop valve considerably improved the rhamnolipid fermentation and significantly enhanced the rhamnolipid productivity by 83% compared to the regular fermentation. This efficient foam breaking was mainly achieved by a high shear rate in combination with fast separation of air from the collapsed foam. Altogether, the stop valve possessed a great activity in breaking rhamnolipid foam, and the involving mechanism holds the potential for developing efficient foam breakers for industrial rhamnolipid fermentation.
Subject(s)
Biotechnology/instrumentation , Biotechnology/methods , Glycolipids/biosynthesis , Equipment Design , Fermentation , Pseudomonas aeruginosa/metabolism , Viscoelastic Substances/chemistryABSTRACT
With recent environmental and health concerns, biosurfactants have obtained increasing interest in replacing conventional surfactants for diverse applications. In agriculture, the use of surfactant in stimulating foliar uptake is mainly for wetting leaf surface, resisting deposition/evaporation, enhancing penetration across cuticular membrane (CM) and translocation. This paper aimed to address the improved foliar uptake by rhamnolipid (RL) in comparison with the currently used alkyl polyglucoside (APG). As found, compared with APG at 900mg/L (1×critical micellar concentration, CMC), RL at a much lower concentration of 50mg/L (1×CMC) showed much better wettability and surface activity, indicative of its high effectiveness as surfactants. Its performance on resistance to deposition and evaporation was at least as same as APG. Moreover, RL could significantly improve the penetration of herbicide glyphosate and other two small water-soluble molecules (phenol red and Fe(2+)) across CM at an equivalent efficiency as APG at 1×CMC. Finally, the greatly enhanced herbicidal actitivity of glyphosate on greenhouse plants confirmed that RL and APG could both enhance the foliar uptake including translocation. Overall, RL should be more applicable than APG in agriculture due to its more promising properties on health/environmental friendliness.
Subject(s)
Glycolipids/pharmacology , Plant Leaves/drug effects , Surface-Active Agents/pharmacology , Chlorophyll/metabolism , Glucosides/chemistry , Glycine/analogs & derivatives , Glycine/toxicity , Iris Plant/chemistry , Phenolsulfonphthalein/pharmacology , Plant Epidermis/drug effects , Plant Leaves/ultrastructure , Solubility , Surface Tension/drug effects , Volatilization , Water/chemistry , Wettability , GlyphosateABSTRACT
In this study, a novel submerged membrane bioreactor (SMBR) with pendulum type oscillation (PTO) hollow fiber membrane modules was developed to treat oily wastewater and control the problem of membrane fouling. To assess the potential of PTO membrane modules, the effect of oscillation orientation and frequency on membrane permeability was investigated in detail. The forces exerted on sludge flocs in the oscillating SMBR were analyzed to evaluate the impact of membrane oscillating on the cake layer resistance reduction. Results showed that the optimized PTO SMBR system exhibited 11 times higher membrane permeability and better fouling controllability than the conventional MBR system. By hydrodynamic analysis, it was found that the cooperative effect of bubble-induced turbulence and membrane oscillation in PTO SMBR system generated strong shear stress at liquid-membrane interface in vertical and horizontal direction and effectively hindered the particles from depositing on membrane surface.
Subject(s)
Biofouling/prevention & control , Bioreactors , Membranes, Artificial , Oils/isolation & purification , Wastewater/chemistry , Water Purification/instrumentation , Water Purification/methods , Biodegradation, Environmental , Biological Oxygen Demand Analysis , Biopolymers/chemistry , Flocculation , Hydrophobic and Hydrophilic Interactions , Particle Size , Permeability , Sewage , Shear Strength , Stress, Mechanical , Time FactorsABSTRACT
Biosurfactant rhamnolipids have been claimed to show biological activities of inhibiting the proliferation of cancer cells. In this study, the cytotoxicity of rhamnolipids was examined on four cancer cells (HepG2, Caco-2, Hela, MCF-7 cells) and two normal cells (HK-2 cell, primary hepatocyte). Interestingly, both cancer cells and normal cells exhibited similar sensitivities to the addition of rhamnolipids in culture medium, and the cytotoxicity was largely attenuated by the presence of fetal bovine serum (FBS) in culture medium. In correlation of the mono-/di-rhamnolipid cytotoxicity with the surface tension of culture medium, it was found that rhamnolipids triggered cytotoxicity whenever the surface tension of culture medium decreased below 41 mN/m irrespective of the FBS content in culture medium, cell line, or rhamnolipid congener. Similarly, each chemical surfactant (Tween-80, sodium dodecyl sulfate, and sodium dodecyl benzene sulfonate) could cause cytotoxicity on HepG2 cells whenever its addition made the surface tension under 41 mN/m in culture medium with or without the presence of FBS. It seems that rhamnolipids, like chemical surfactants, exhibited cytotoxicity by reducing the surface tension of culture medium rather than by changing its specific molecular structure, which had no selection on tumor cells. This study could offer helps to correct the misleading biological activity of rhamnolipids and to avoid the possible large wastes of time and expenses on developing the applications in antitumor drugs.
Subject(s)
Culture Media/chemistry , Glycolipids/toxicity , Surface Tension , Surface-Active Agents/toxicity , Cell Line , Cell Survival/drug effects , HumansABSTRACT
Oily sludge dewatering is practically needed prior to sludge treatments. However, the conventional use of physical treatments with or without chemical conditionings presented poor feasibility in industrial applications due to either poor cost-efficiency or lacking environmental friendliness. In this paper, biosurfactant rhamnolipid was for the first time applied for dewatering of oily sludge. Rhamnolipid treatments under the concentration of 300-1000 mg/L, pH of 5-7 and temperature of 10-60 °C could directly separate 50-80% of water from the stable oily sludge. And both mono-rhamnolipid and di-rhamnolipid were identified to be of equivalent dewatering ability, which is closely related to their equivalent performance in breaking the emulsified oil droplets. Demulsification was found to be involved in settling water from oily sludge. Furthermore, the effectiveness of rhamnolipid was further demonstrated at pilot scale (1000 L) treatment of oily sludge. After pilot treatment, the settled water with residual oil of 10 mg/L and soluble COD of about 800 mg/L could be directly effluxed into the biotreatment system while the concentrated oil sludge with a reduced volume by 60-80% can be pumped into coking tower, achieving completely harmless treatment. It seems that rhamnolipid as dewatering agent was of great prospects in the industrial dewatering of oily sludge.
Subject(s)
Glycolipids/chemistry , Oils/chemistry , Sewage/chemistry , Water Purification , Emulsions/chemistry , Hydrogen-Ion Concentration , Pilot Projects , Surface-Active Agents/chemistry , TemperatureABSTRACT
This work aimed to evaluate the applicability of rhamnolipids as permeation enhancers for oral drugs. In this study, rhamnolipids were found to effectively increase the paracellular and transcellular transport of Transwell(®)-cultured Caco-2 cells, an in vitro model of the human small intestinal epithelium, in a concentration-dependent manner. Rhamnolipids at 150 mg/L increased the paracellular apparent permeability (Papp) of phenol red almost 7- to 8-fold, the largest enhancement ever reported, while Tween-80 exhibited no such effect. Regarding the transcellular pathway, rhamnolipids at 150 mg/L enhanced the Papp of propranolol 2-fold, similar to the performance of Tween-80 at 400mg/L. Moreover, rhamnolipids like Tween-80, significantly inhibited P-glycoprotein (P-gp) activity reflected by the reduced efflux ratio (basolateral-to-apical/apical-to-basolateral) of rhodamine 123 (R123), a P-gp substrate, on Caco-2 cells. Inhibition of P-gp activity was confirmed on plate cultured Caco-2 monolayers by assaying accumulation/efflux of R123 and R110, a non-P-gp substrate. Finally, rhamnolipids were demonstrated to be safe by cell viability and hemolysis assays. In conclusion, rhamnolipids were highly effective regulators of all three transport pathways, suggesting their use as a safe absorption enhancer for oral drugs.
Subject(s)
Glycolipids/pharmacology , Intestinal Absorption , Animals , Biological Transport/drug effects , Caco-2 Cells , Cell Membrane Permeability/drug effects , Cell Survival/drug effects , Cells, Cultured , Erythrocytes/drug effects , Erythrocytes/pathology , Fluorescent Dyes/metabolism , Hemolysis , Humans , Intestinal Mucosa/metabolism , Permeability/drug effects , Rats , Rhodamine 123/metabolism , Rhodamines/metabolismABSTRACT
Waste crude oil emulsion was inevitably produced in the petroleum industrial process, causing harmful impact on the ecological and social environment. In this study, rhamnolipid was for the first time investigated for demulsification of waste crude oil. As found in this paper, rhamnolipid treatment could obtain over 90% of dewatering efficiency on refractory waste crude oil and such efficient demulsification was confirmed on model emulsions. As further demonstrated on the pilot scale (100 L), rhamnolipid treatment could recover over 98% of crude oil from the wastes. The recovered oil contained less than 0.3% of water and thus can directly re-enter into refinery process while the aqueous phase can be disposed into dischargeable water due to largely reduced soluble COD after subjected to 5 days of active sludge treatment. It seems that rhamnolipids as biodemulsifiers were of great prospects in the industrial demulsification of waste crude oil.
Subject(s)
Conservation of Natural Resources/methods , Emulsions/chemistry , Excipients/chemistry , Glycolipids/chemistry , Industrial Waste/prevention & control , Petroleum/analysis , Waste Disposal, Fluid/methods , Biodegradation, Environmental , Pilot ProjectsABSTRACT
Oily wastewater generated by various industries creates a major ecological problem throughout the world. The traditional methods for the oily wastewater treatment are inefficient and costly. Surfactants can promote the biodegradation of petroleum hydrocarbons by dispersing oil into aqueous environment. In the present study, we applied rhamnolipid-containing cell-free culture broth to enhance the biodegradation of crude oil and lubricating oil in a conventional aerobically-activated sludge system. At 20 degrees C, rhamnolipids (11.2 mg/L) increased the removal efficiency of crude oil from 17.7% (in the absence of rhamnolipids) to 63%. At 25 degrees C, the removal efficiency of crude oil was over 80% with the presence of rhamnolipids compared with 22.3% in the absence of rhamnolipids. Similarly, rhamnolipid treatment (22.5 mg/L) for 24 h at 20 degrees C significantly increased the removal rate of lubricating oil to 92% compared with 24% in the absence of rhamnolipids. The enhanced removal of hydrocarbons was mainly attributed to the improved solubility and the reduced interfacial tension by rhamnolipids. We conclude that a direct application of the crude rhamnolipid solution from cell culture is effective and economic in removing oily contaminants from wastewater.