Rhamnolipids are glycolipidic biosurfactants produced by various bacterial species. They were initially found as exoproducts of the opportunistic pathogen Pseudomonas aeruginosa and described as a mixture of four congeners: α-L-rhamnopyranosyl-α-L-rhamnopyranosyl-β-hydroxydecanoyl-β-hydroxydecanoate (Rha-Rha-C10-C10), α-L-rhamnopyranosyl-α-L-rhamnopyranosyl-β-hydroxydecanoate (Rha-Rha-C10), as well as their mono-rhamnolipid congeners Rha-C10-C10 and Rha-C10. The development of more sensitive analytical techniques has lead to the further discovery of a wide diversity of rhamnolipid congeners and homologues (about 60) that are produced at different concentrations by various Pseudomonas species and by bacteria belonging to other families, classes, or even phyla. For example, various Burkholderia species have been shown to produce rhamnolipids that have longer alkyl chains than those produced by P. aeruginosa. In P. aeruginosa, three genes, carried on two distinct operons, code for the enzymes responsible for the final steps of rhamnolipid synthesis: one operon carries the rhlAB genes and the other rhlC. Genes highly similar to rhlA, rhlB, and rhlC have also been found in various Burkholderia species but grouped within one putative operon, and they have been shown to be required for rhamnolipid production as well. The exact physiological function of these secondary metabolites is still unclear. Most identified activities are derived from the surface activity, wetting ability, detergency, and other amphipathic-related properties of these molecules. Indeed, rhamnolipids promote the uptake and biodegradation of poorly soluble substrates, act as immune modulators and virulence factors, have antimicrobial activities, and are involved in surface motility and in bacterial biofilm development.
The nonconventional, oleaginous yeast, Yarrowia lipolytica is rapidly emerging as a valuable host for the production of a variety of both lipid and nonlipid chemical products. While the unique genetics of this organism pose some challenges, many new metabolic engineering tools have emerged to facilitate improved genetic manipulation in this host. This review establishes a case for Y. lipolytica as a premier metabolic engineering host based on innate metabolic capacity, emerging synthetic tools, and engineering examples. The metabolism underlying the lipid accumulation phenotype of this yeast as well as high flux through acyl-CoA precursors and the TCA cycle provide a favorable metabolic environment for expression of relevant heterologous pathways. These properties allow Y. lipolytica to be successfully engineered for the production of both native and nonnative lipid, organic acid, sugar and acetyl-CoA derived products. Finally, this host has unique metabolic pathways enabling growth on a wide range of carbon sources, including waste products. The expansion of carbon sources, together with the improvement of tools as highlighted here, have allowed this nonconventional organism to act as a cellular factory for valuable chemicals and fuels.
Physical and chromatographic characterization of the surfactin biosurfactant produced by Bacillus subtilis isolate BS5 has been conducted to study its potentiality for industrial application. The crude extract of test surfactin appeared as off-white to buff flake-like amorphous residue with bad odor similar to sour pomegranate. Test surfactin showed solubility in aqueous solution at pH>5 with optimum solubility at pH 8-8.5. It was also soluble in organic solvents like ethanol, acetone, methanol, butanol, chloroform, and dichloromethane. Surfactin crystals appeared rectangular with blunt corners and were arranged perpendicular to each other making a plus sign. Extracted surfactin showed high surface activity, as it could lower the surface tension of water from about 70 to 36 mN/m at approximately 15.6 mg/l. Moreover, test surfactin exhibited excellent stabilities at high temperatures (100 degrees C for up to 1 h at and autoclaving at 121 degrees C for 10 min), salinities (up to 6% NaCl), and over a wide range of pH (5-13). Test surfactin in the cell-free supernatant or crude culture broth forms showed high emulsification indices against kerosene (62.5% and 59%, respectively), diesel (62.5% and 66%, respectively), and motor oil (62% and 66%, respectively). These characters can effectively make test surfactin, in its crude forms, a potential candidate for the use in bioremediation of hydrocarbon-contaminated sites or in the petroleum industry. Chromatographic characterization of test surfactin, using high-performance liquid chromatography technique, revealed that the extracted surfactin contained numerous isoforms, of which six were found in the standard surfactin preparation (Fluka). Additional peaks appeared in the test surfactin and not in the standard one. These peaks may correspond to new surfactin isoforms that may be present in the test surfactin produced by B. subtilis isolate BS5.
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