Antibiofilm and wound healing efficacy of rhamnolipid biosurfactant against pathogenic bacterium Staphylococcus aureus.

The present study evaluates the in-vitro antibiofilm activity against the biofilm formed by Staphylococcus aureus, and the wound-healing efficacy of two different types of rhamnolipids produced by Pseudomonas aeruginosa strain JS29 in S.aureus infected wounds. The biosurfactant production was carried out in a mineral salt medium supplemented with 2 % Glucose and 2 % Glycerol individually and thus were designated as RL-Glu and RL-Gly respectively. 0.5 mg/ml of RL-Glu and RL-Gly demonstrated 90 % growth inhibition of S. aureus while exhibiting bactericidal activity at 4 mg/ml of RL-Glu and 1 mg/ml of RL-Gly. Both types of rhamnolipid cause changes in membrane permeability leading to pathogens' non-viability. 90 % inhibition of biofilm formation by S. aureus was observed at 2 mg/ml of RL-Glu and 0.5 mg/ml of RL-Gly, while 0.5 mg/ml of both rhamnolipid disrupted 90 % of the preformed biofilm. 0.5 mg/ml of RL-Glu and RL-Gly decreases the production of exopolysaccharides and also causes structural alteration. 0.5 mg/ml of RL-Glu and RL-Gly were found to exhibit effective wound healing efficacy in S. aureus infected wounds within 7 days of treatment. Histopathological studies of wound sites revealed efficient wound management by both the rhamnolipid. LCMS and GCMS characterization of the biosurfactant revealed that JS29 produces different rhamnolipid congeners when grown on different carbon sources, thereby influencing the antimicrobial, antibiofilm, and wound healing efficacy of rhamnolipid.

Performance evaluation of rhamnolipid biosurfactant produced by Pseudomonas aeruginosa and its effect on marine oil-spill remediation.

This study aimed to reveal that the effect of biosurfactant on the dispersion and degradation of crude oil. Whole genome analysis showed that Pseudomonas aeruginosa GB-3 contained abundant genes involved in biosurfactant synthesis and metabolic processes and had the potential to degrade oil. The biosurfactant produced by strain GB-3 was screened by various methods. The results showed that the surface tension reduction activity was 28.6 mN·m-1 and emulsification stability was exhibited at different pH, salinity and temperature. The biosurfactant was identified as rhamnolipid by LC-MS and FTIR. The fermentation conditions of strain GB-3 were optimized by response surface methodology, finally the optimal system (carbon source: glucose, nitrogen source: ammonium sulfate, C/N ratio:16:1, pH: 7, temperature: 30-35 °C) was determined. Compared with the initial fermentation, the yield of biosurfactant increased by 4.4 times after optimization. In addition, rhamnolipid biosurfactant as a dispersant could make the dispersion of crude oil reach 38% within seven days, which enhanced the bioavailability of crude oil. As a biostimulant, it could also improve the activity of indigenous microorganism and increase the degradation rate of crude oil by 10-15%. This study suggested that rhamnolipid biosurfactant had application prospect in bioremediation of marine oil-spill.

Biosurfactant-capped CuO nanoparticles coated cotton/polypropylene fabrics toward antimicrobial textile applications.

The global COVID-19 pandemic has led to an increase in the importance of implementing effective measures to prevent the spread of microorganisms. Consequently, there is a growing demand for antimicrobial materials, specifically antimicrobial textiles and face masks, because of the surge in diseases caused by bacteria and viruses like SARS-CoV-2. Face masks that possess built-in antibacterial properties can rapidly deactivate microorganisms, enabling reuse and reducing the incidence of illnesses. Among the numerous types of inorganic nanomaterials, copper oxide nanoparticles (CuO NPs) have been identified as cost-effective and highly efficient antimicrobial agents for inactivating microbes. Furthermore, biosurfactants have recently been recognized for their potential antimicrobial effects, in addition to inorganic nanoparticles. Therefore, this research's primary focus is synthesizing biosurfactant-mediated CuO NPs, integrating them into natural and synthetic fabrics such as cotton and polypropylene and evaluating the resulting fabrics' antimicrobial activity. Using rhamnolipid (RL) as a biosurfactant and employing a hydrothermal method with a pH range of 9-11, RL-capped CuO NPs are synthesized (RL-CuO NPs). To assess their effectiveness against gram-positive (Staphylococcus aureus) and gram-negative (Escherichia coli) microorganisms, the RL-CuO NPs are subjected to antibacterial testing. The RL-capped CuO NPs exhibited antimicrobial activity at much lower concentrations than the individual RL, CuO. RL-CuO NPs have shown a minimum inhibitory concentration (MIC) of 1.2 mg ml-1and minimum bactericidal concentration (MBC) of 1.6 mg ml-1forE. coliand a MIC of 0.8 mg ml-1and a MBC of 1.2 mg ml-1forS. aureus, respectively. Furthermore, the developed RL-CuO NPs are incorporated into cotton and polypropylene fabrics using a screen-printing technique. Subsequently, the antimicrobial activity of the coated fabrics is evaluated, revealing that RL-CuO NPs coated fabrics exhibited remarkable antibacterial properties against both gram-positive and gram-negative bacteria.

Common operational issues and possible solutions for sustainable biosurfactant production from lignocellulosic feedstock.

Surfactants are compounds with high surface activity and emulsifying property. These compounds find application in food, medical, pharmaceutical, and petroleum industries, as well as in agriculture, bioremediation, cleaning, cosmetics, and personal care product formulations. Due to their widespread use and environmental persistence, ensuring biodegradability and sustainability is necessary so as not to harm the environment. Biosurfactants, i.e., surfactants of plant or microbial origin produced from lignocellulosic feedstock, perform better than their petrochemically derived counterparts on the scale of net-carbon-negativity. Although many biosurfactants are commercially available, their high cost of production justifies their application only in expensive pharmaceuticals and cosmetics. Besides, the annual number of new biosurfactant compounds reported is less, compared to that of chemical surfactants. Multiple operational issues persist in the biosurfactant value chain. In this review, we have categorized some of these issues based on their relative position in the value chain - hurdles occurring during planning, upstream processes, production stage, and downstream processes - alongside plausible solutions. Moreover, we have presented the available paths forward for this industry in terms of process development and integrated pretreatment, combining conventional tried-and-tested strategies, such as reactor designing and statistical optimization with cutting-edge technologies including metabolic modeling and artificial intelligence. The development of techno-economically feasible biosurfactant production processes would be instrumental in the complete substitution of petrochemical surfactants, rather than mere supplementation.

Valorization of waste engine oil to mono- and di-rhamnolipid in a sustainable approach to circular bioeconomy.

This study aims to valorize waste engine oil (WEO) for synthesizing economically viable biosurfactants (rhamnolipids) to strengthen the circular bioeconomy concept. It specifically focuses on investigating the influence of key bioprocess parameters, viz. agitation and aeration rates, on enhancing rhamnolipid yield in a fed-batch fermentation mode. The methodology involves conducting experiments in a stirred tank bioreactor (3 L) using Pseudomonas aeruginosa gi |KP 163922| as the test organism. Central composite design and response surface methodology (CCD-RSM) are employed to design the experiments and analyze the effects of agitation and aeration rates on various parameters, including dry cell biomass (DCBM), surface tension, tensoactivity, and rhamnolipid yield. It is also essential to determine the mechanistic pathway of biosurfactant production followed by the strain using complex hydrophobic substrates such as WEO. The study reveals that optimal agitation and aeration rates of 200 rpm and 1 Lpm result in the highest biosurfactant yield of 29.76 g/L with minimal surface tension (28 mN/m). Biosurfactant characterization using FTIR, 1H NMR, and UPLC-MS/MS confirm the presence of dominant molecular ion peaks m/z 543.9 and 675.1. This suggests that the biosurfactant is a mixture of mono- and di-rhamnolipids (RhaC10C10, RhaRhaC10C12:1, RhaRhaC12:1C10). The findings present a sustainable approach for biosurfactant production in a fed-batch bioreactor. This research opens the possibility of exploring the use of pilot or large-scale bioreactors for biosurfactant production in future investigations.

The efficiency of micellar solubilization of naphthalene from aqueous solutions using rhamnolipid as a biological surfactant according to NMR diffusometry.

The micellar solubilization of naphthalene from its saturated aqueous solutions using the biosurfactant rhamnolipid was studied. Using the NMR diffusion method, selective measurements of the self-diffusion coefficients of molecules of all components of the solution-naphthalene, rhamnolipid, and water-were carried out at various rhamnolipid concentrations from 0.06 to 100 g/L. Based on the results of diffusometry, the distribution of naphthalene molecules between the states free in solution and states bound by micelles was found. With an increase in the concentration of rhamnolipids, the proportion of bound naphthalene molecules increases from 50% at CRL = 2 g/L to 100% at CRL ≥ 50 g/L. The micelle-water partition coefficient Km and the molar solubilization ratio MSR were calculated.

A Rare Mono-Rhamnolipid Congener Efficiently Produced by Recombinant Pseudomonas aeruginosa YM4 via the Expression of Global Transcriptional Regulator irrE.

Rhamnolipids (RLs) are widely used biosurfactants produced mainly by Pseudomonas aeruginosa and Burkholderia spp. in the form of mixtures of diverse congeners. The global transcriptional regulator gene irrE from radiation-tolerant extremophiles has been widely used as a stress-resistant element to construct robust producer strains and improve their production performance. A PrhlA-irrE cassette was constructed to express irrE genes in the Pseudomonas aeruginosa YM4 of the rhamnolipids producer strain. We found that the expression of irrE of Deinococcus radiodurans in the YM4 strain not only enhanced rhamnolipid production and the strain's tolerance to environmental stresses, but also changed the composition of the rhamnolipid products. The synthesized rhamnolipids reached a maximum titer of 26 g/L, about 17.9% higher than the original, at 48 h. The rhamnolipid production of the recombinant strain was determined to be mono-rhamnolipids congener Rha-C10-C12, accounting for 94.1% of total products. The critical micelle concentration (CMC) value of the Rha-C10-C12 products was 62.5 mg/L and the air-water surface tension decreased to 25.5 mN/m. The Rha-C10-C12 products showed better emulsifying activity on diesel oil than the original products. This is the first report on the efficient production of the rare mono-rhamnolipids congener Rha-C10-C12 and the first report that the global regulator irrE can change the components of rhamnolipid products in Pseudomonas aeruginosa.

Optimizing Systems for Robust Heterologous Production of Biosurfactants Rhamnolipid and Lyso-Ornithine Lipid in Pseudomonas putida KT2440.

Surfactants are amphiphilic molecules that are capable of mixing water and oil. Biosurfactants are eco-friendly, low-toxicity, and stable to a variety of environmental factors. Optimizing conditions for microorganisms to produce biosurfactants can lead to improved production suitable for scaling up. In this study, we compared heterologous expression levels of the luminescence system luxCDABE operon controlled by regulatable promoters araC-PBAD and its strong version araC-PBAD-SD in Escherichia coli K12, Pseudomonas aeruginosa PAO1, and P. putida KT2440. Real-time monitoring of luminescence levels in the three strains indicated that luxCDABE controlled by araC-PBAD-SD promoter with 0.2% arabinose supplementation in P. putida produced the highest level of luminescence. By using the araC-PBAD-SD promoter-controlled rhlAB expression in P. putida, we were able to produce mono-rhamnolipid at a level of 1.5 g L-1 when 0.02% arabinose was supplemented. With the same system to express olsB, lyso-ornithine lipid was produced at a level of 10 mg L-1 when 0.2% arabinose was supplemented. To our knowledge, this is the first report about optimizing conditions for lyso-ornithine lipid production at a level up to 10 mg L-1. Taken together, our results demonstrate that regulatable araC-PBAD-SD promoter in P. putida KT2440 is a useful system for heterologous production of biosurfactants.

Effects of rhamnolipid replacement of chlortetracycline on growth performance, slaughtering traits, meat quality and antioxidant function in broilers.

BACKGROUND: Rhamnolipids (RLS) are surfactants that improve the growth performance of poultry by improving the absorption of nutrients. This study aims to investigate the effects of RLS replacement of chlortetracycline (CTC) on growth performance, slaughtering traits, meat quality, antioxidant function and nuclear-factor-E2-related factor 2 (Nrf2) signaling pathway in broilers. A total of 600 one-day-old Arbor Acres chicks were randomly assigned to five groups with eight replicates in each group, raised for 42 days. Broilers were respectively fed a basal diet with no CTC or RLS, 75 mg kg-1 CTC, and 250, 500, 1000 mg kg-1 RLS. RESULTS: Dietary supplementation with RLS linearly increased the average daily gain, average daily feed intake, carcass yield, eviscerated yield, ether extract, enhanced total superoxide and glutathione peroxidase (GPx) activities, overexpressed the relative expressions of Nrf2, heme oxygenase 1, Copper/zinc superoxide dismutase, Manganese superoxide dismutase, GPx and catalase and decreased the lightness value at 24 h, drip loss and malondialdehyde contents of broilers (P < 0.05). Compared with the control group, broilers fed 1000 mg kg-1 RLS reduced the drip loss and broilers fed 500 mg kg-1 RLS increased muscle crude fat content (P < 0.05). Compared with the CTC group, dietary supplementation with 1000 mg kg-1 RLS increased eviscerated yield (P < 0.05). CONCLUSION: RLS could improve growth performance, crude fat content, meat quality and antioxidant capacity and activate relative expression of genes in the Nrf2 signaling pathway in broilers. It could be used as an antibiotic substitute in diets and the recommended supplemental dose of RLS in feed of broilers is 1000 mg kg-1. © 2024 Society of Chemical Industry.

Investigation of Photodynamic and Rhamnolipid Inhibition on the Dermatophyte Biofilm.

The failure to successfully treat dermatophyte-related diseases is often due to the formation of biofilms, which makes dermatophytes resistant to antifungals. Here, an attempt has been made to assess inhibition of dermatophyte biofilm production using photodynamic therapy and rhamnolipid biosurfactant. Two methods were used to inhibit biofilm formation by dermophytes Trichophyton mentagrophytes, Trichophyton rubrum and Trichophyton verrucosum, Microsporum canis and Microsporum gypseum. The first method was the use of rhamnolipid with concentrations of 39 to 1000 ppm and the second was the use of photodynamic method with concentrations of 8, 16 and 32 µg/ml of methylene blue. In addition, these two methods were evaluated simultaneously. The biofilm formation was evaluated using spectrophotometry and scanning electron microscopy. Biosurfactant has been shown to have an improved ability to inhibit the formation of biofilm by the strains. Although photodynamic therapy has not been successful, but in combination with biosurfactant, it may have a synergistic effect. By investigating the effect of rhamnolipid on the formation of biofilm, it was found that Microsporum species has a relatively stronger attachment to the surfaces of the wells compared to trichophyton species. The biofilms were evaluated with electron microscope in the simultaneous treatment of rhamnolipid and photodynamics. The results showed that after the treatment, the biofilms became discrete and their structural integrity was reduced. Even in Microsporum species, which were among the most resistant dermatophytes, the changes in the fungal biofilm after treatment were significant.

Improved phyllosphere microbiome composition of tea plant with the application of small peptides in combination with rhamnolipid.

BACKGROUND: Small peptides play a crucial role in plant growth and adaptation to the environment. Exogenous small peptides are often applied together with surfactants as foliar fertilizers, but the impact of small peptides and surfactants on the tea phyllosphere microbiome remains unknown. RESULTS: In this study, we investigated the effects of small peptides and different surfactants on the tea phyllosphere microbiome using 16S and ITS sequencing. Our results showed that the use of small peptides reduced the bacterial diversity of the tea phyllosphere microbiome and increased the fungal diversity, while the use of surfactants influenced the diversity of bacteria and fungi. Furthermore, the addition of rhamnolipid to small peptides significantly improved the tea phyllosphere microbiome community structure, making beneficial microorganisms such as Pseudomonas, Chryseobacterium, Meyerozyma, and Vishniacozyma dominant populations. CONCLUSION: Our study suggests that the combined use of small peptides and surfactants can significantly modify the tea phyllosphere microbiome community structure, particularly for beneficial microorganisms closely related to tea plant health. Thus, this preliminary study offers initial insights that could guide the application of small peptides and surfactants in agricultural production, particularly with respect to their potential for modulating the phyllosphere microbiome community in tea plant management.

Assessment of synergistic activity of rhamnolipid and linezolid against methicillin-resistant Staphylococcus aureus in-vitro and in-vivo with Galleria mellonella larvae model.

Antibiotic resistance, one of the most crucial public health problems, has increased the interest in synergy studies of antibiotics with existing antibiotics and natural compounds to make current treatment more effective in addition to new drug development. In this study, the effectiveness of rhamnolipid and linezolid on the Galleria mellonella larvae model in-vitro and in-vivo against Methicillin-resistant Staphylococcus aureus isolates, which are problematic in treatment, were investigated. Four S.aureus (One ATCC 29213 strain and three methicillin-resistant strains) were used in the study. Two MRSA isolates were resistant to linezolid, and one was susceptible. Partial synergy was observed in one resistant strain, and although no synergy was observed in the other resistant strain, the minimum inhibitory concentration of the resistant strain decreased from 16 to 4 µg/mL with a four-fold decrease and reached the susceptibility limit. No change was observed in the MIC of linezolid-susceptible strains. The G.mellonella larval model demonstrated that combined therapy was more effective than monotherapy by survival function tests and CFU determination. RML/LNZ combination improved survival compared to monotherapy and decreased the bacterial burden from 108 to 103.

Wound healing efficacy of rhamnolipid-coated zinc oxide nanoparticle along with its in vivo antibacterial efficacy against Staphylococcus aureus.

Rhamnolipids are microbial metabolites with antibacterial efficacies, which can be further boosted through the application of nanobiotechnology. In this study, the efficacy of rhamnolipid-coated zinc oxide nanoparticles (ZnRL) has been studied for their wound healing efficacy as well as in vivo antibacterial efficacy. Thus, this study evaluates the efficacy of ZnRL to heal an excised infected wound, which was compared with the healing efficacy of rhamnolipid and clindamycin. The study revealed that rhamnolipid-coated zinc oxide nanoparticles possess promising wound healing efficacy with prominent antibacterial activity in the rat model. Prominent wound healing in a Staphylococcus aureus infected excised wound was observed on the 5th day of the treatment when the wound site was treated with 100 µl of 0.5 mg/ml of ZnRL. This concentration of ZnRL was found to exhibit efficient antibacterial activity against the pathogen, thereby decreasing the amount of pathogen in the wound site. ZnRL exhibited efficient wound contraction, thereby decreasing the size of the wound prominently in 5 days. Histological study revealed efficient tissue remodelling in ZnRL-treated skin which resulted in rapid formation of the epidermis and recruitment of various dermal cells within the 5th day of treatment. The study also revealed the non-cytotoxic effect of the nanoparticles in fibroblast cell line L929 and the non-haemolytic effect against blood cells, indicating its potential in pharmaceuticals.

A novel membrane stirrer system enables foam-free biosurfactant production.

Bioreactors are the operative backbone, for example, for the production of biopharmaceuticals, biomaterials in tissue engineering, and sustainable substitutes for chemicals. Still, the Achilles' heel of bioreactors nowadays is the aeration which is based on intense stirring and gas sparging, yielding inherent drawbacks such as shear stress, foaming, and sterility concerns. We present the synergistic combination of simulations and experiments toward a membrane stirrer for the efficient bubble-free aeration of bioreactors. A digital twin of the bioreactor with an integrated membrane-module stirrer (MemStir) was developed with computational fluid dynamics (CFD) studies addressing the determination of fluid mixing, shear rates, and local oxygen concentration. Usability of the MemStir is shown in a foam-free recombinant production process of biosurfactants (rhamnolipids) from glucose with different strains of Pseudomonas putida KT2440 in a 3-L vessel and benchmarked against a regular aerated process. The MemStir delivered a maximal oxygen transfer rate (OTRmax ) of 175 mmol L-1 h-1 in completely foam-free cultivations. With a high space-time yield (STY) of 118 mgRL L-1 h-1 during a fed-batch fermentation, the effectiveness of the novel MemStir is demonstrated. Simulations show the generic value of the MemStir beyond biosurfactant production, for example, for animal cell cultivation.

Synthesis of magnetically recyclable porous cross-linked aggregates of Tramates versicolor MTCC 138 laccase for the efficient removal of pentachlorophenol from aqueous solution.

The primary objective of this study is to synthesize the magnetically separable highly active porous immobilized laccase for the removal of pentachlorophenol (PCP) in an aqueous solution. Magnetic porous cross-linked enzyme aggregates (Mp-CLEAs) of laccase were synthesized using 1% starch solution with 5 mM glutaraldehyde followed by 10 h of cross-linking time with an activity recovery of 90.85 ± 0.2%. The biocatalytic efficiency of magnetic porous CLEAs (Mp-CLEAs) was 2-fold higher than that of magnetic CLEAs. The synthesized Mp-CLEAs were mechanically stable with enhanced catalytic efficiency, and reusability thus overcoming the mass transfer limitations and enzyme loss. At 40 °C, the thermal stability of the magnetic porous immobilized laccase was improved, with a 602 min half-life compared to 207 min half-life for the free enzyme. Using 40 U/mL of laccase for the removal of 100 ppm of PCP, M-CLEAs, and Mp-CLEAs removed 60.44% and 65.53% of PCP, respectively. Furthermore, to enhance PCP removal, a laccase-aided system was harnessed by optimizing various surfactants and mediators. Of these, 0.1 mM of rhamnolipid and 2,3 dimethoxy phenol had the highest PCP removal rates of 95.12% and 99.41%, respectively, for Mp-CLEAs. This study demonstrates the efficacy of the laccase-surfactant-mediator system for the removal of PCP from the aqueous solution, which can also be proposed for real-time application.

Quantitative determination of rhamnolipid using HPLC-UV through carboxyl labeling.

Rhamnolipid, as a low-toxic, biodegradable and environmentally friendly biosurfactant, has broad application prospects in many industries. However, the quantitative determination of rhamnolipid is still a challenging task. Here, a new sensitive method for the quantitative analysis of rhamnolipid based on a simple derivatization reaction was developed. In this study, 3-[3'-(l-rhamnopyranosyloxy) decanoyloxy] decanoic acid (Rha-C10-C10) and 3-[3'-(2'-O-α-l-rhamnopyranosyloxy) decanoyloxy] decanoic acid (Rha-Rha-C10-C10) were utilized as the representative rhamnolipids. Liquid chromatography-mass spectrometry and high-performance liquid chromatography-ultra violet results showed that these two compounds were successfully labeled with 1 N1-(4-nitrophenyl)-1,2-ethylenediamine. There was an excellent linear relationship between rhamnolipid concentration and peak area of labeled rhamnolipid. The detection limits of the Rha-C10-C10 and Rha-Rha-C10-C10 were 0.018 mg/L (36 nmol/L) and 0.014 mg/L (22 nmol/L), respectively. The established amidation method was suitable for the accurate analysis of rhamnolipids in the biotechnological process. The method had good reproducibility with the relative standard deviation of 0.96% and 0.79%, respectively, and sufficient accuracy with a recovery of 96%-100%. This method was applied to quantitative analysis of 10 rhamnolipid homologs metabolized by Pseudomonas aeruginosa LJ-8. The single labeling method was used for the quantitative analysis of multiple components, which provided an effective method for the quality evaluation of other glycolipids with carboxyl groups.

Antimicrobial, antiadhesive, and antibiofilm actions of rhamnolipids on ESKAPE pathogens.

The aim of this study was to test the antimicrobial, antiadhesive, and antibiofilm activities of a rhamnolipid extracted from Pseudomonas aeruginosa UKMP14T previously isolated from oil-contaminated soil in Malaysia against ESKAPE (i.e. multidrug resistant) pathogens. Zones of inhibition in an agar well diffusion assay were observed at 50 µg mL-1 concentrations of rhamnolipid for all the ESKAPE bacteria. The MIC and MBC values ranged between 7.81-62.5 µg mL-1 and 31.25-1000 µg mL-1, respectively. Percent killing was recorded to be >90% except for Klebsiella pneumoniae (86.84%). Furthermore, antiadhesion studies showed that there was 76% hindrance in attachment of Enterococcus faecium and 91% in Acinetobacter baumannii at 4 × MIC. The highest inhibition in adhesion was found at 4 × MIC, which was 46% for Ac. baumannii and 62% for Enterococcus faecium. Finally, the antibiofilm capability of the rhamnolipid was determined, which ranged between 25%-76% in Ac. baumannii and 35%-88% in Enterococcus faecium. To the best of our knowledge, this is the first study to include research on antimicrobial, antiadhesive and antibiofilm activities of rhamnolipid from the local isolate Ps. aeruginosa UKMP14T against ESKAPE bacteria. Obtained results suggest that this rhamnolipid can be exploited commercially for the production of novel antibiotics.

Extended Biosynthesis of Rhamnolipid by Immobilized Pseudomonas aeruginosa USM-AR2 Cells in a Fluidized Bed Bioreactor.

This study aims to evaluate rhamnolipid production by immobilized Pseudomonas aeruginosa USM-AR2 cells using waste cooking oil (WCO) as the carbon source. P. aeruginosa USM-AR2 cells were entrapped in polyvinyl alcohol (PVA)-alginate hydrogel beads. The performance of entrapped cells was compared with free cells in shake flasks before cultivation in a custom-designed fluidized bed reactor (FBR). A mass of 1 g wet cells entrapped in PVA-alginate hydrogel beads were successfully recycled 3 times in shake flasks at 200 rpm, producing between 0.66-1.34 g L-1 rhamnolipid after 120 h. Meanwhile, cultivation of entrapped cells in FBR with broth recirculation showed that the suitable hydrogel beads to medium ratio was 1:20 ratio at an aeration rate of 0.5 vvm, producing between 0.77 to 1.58 g L-1 rhamnolipid, degrading 8.67 to 20.93 g L-1 of waste cooking oil in 15 cycles of repeated batch cultivation. Entrapped P. aeruginosa USM-AR2 cells achieved a higher rhamnolipid production, by 1.03-fold during cycle 3 in shake flasks and 1.19-fold during cycle 11 in an FBR, compared to free cells. These results show that entrapment enables reusability of viable cells and maintains stability of rhamnolipid production throughout the extended cultivation, increasing cell tolerance to perturbations in fermentation conditions.

Rhamnolipid Self-Aggregation in Aqueous Media: A Long Journey toward the Definition of Structure-Property Relationships.

The need to protect human and environmental health and avoid the widespread use of substances obtained from nonrenewable sources is steering research toward the discovery and development of new molecules characterized by high biocompatibility and biodegradability. Due to their very widespread use, a class of substances for which this need is particularly urgent is that of surfactants. In this respect, an attractive and promising alternative to commonly used synthetic surfactants is represented by so-called biosurfactants, amphiphiles naturally derived from microorganisms. One of the best-known families of biosurfactants is that of rhamnolipids, which are glycolipids with a headgroup formed by one or two rhamnose units. Great scientific and technological effort has been devoted to optimization of their production processes, as well as their physicochemical characterization. However, a conclusive structure-function relationship is far from being defined. In this review, we aim to move a step forward in this direction, by presenting a comprehensive and unified discussion of physicochemical properties of rhamnolipids as a function of solution conditions and rhamnolipid structure. We also discuss still unresolved issues that deserve further investigation in the future, to allow the replacement of conventional surfactants with rhamnolipids.

Holistic approach to waste mobil oil bioremediation: Valorizing waste through biosurfactant production for soil restoration.

The combustion of mobil oil leads to the emission of toxic compounds in the environment. In this study, the aromatic and aliphatic hydrocarbon fractions present in a waste mobil oil collected from automobile market were comprehensively identified and their toxicity was evaluated using wheat grain. Lysinibacillus sphaericus strain IITR51 isolated and characterized previously could degrade 30-80% of both aliphatic and aromatic hydrocarbons in liquid culture. Interestingly, the strain IITR51 produced 627 mg/L of rhamnolipid biosurfactant by utilizing 3% (v/v) of waste mobil oil in the presence of 1.5% glycerol as additional carbon source. In a soil microcosm study by employing strain IITR51, 50-86% of 3-6 ring aromatic hydrocarbons and 63-98% of aliphatic hydrocarbons (C8 to C22) were degraded. Addition of 60 µg/mL rhamnolipid biosurfactant enhanced the degradation of both aliphatic and aromatic hydrocarbons from 76.88% to 61.21%-94.11% and 78.27% respectively. The degradation of mobil oil components improved the soil physico-chemical properties and increased soil fertility to 64% as evident by the phytotoxicity assessments. The findings indicate that strain IITR51 with degradation capability coupled with biosurfactant production could be a candidate for restoring hydrocarbon contaminated soils.