Phase Behaviour, Functionality, and Physicochemical Characteristics of Glycolipid Surfactants of Microbial Origin.

Growing demand for biosurfactants as environmentally friendly counterparts of chemically derived surfactants enhances the extensive search for surface-active compounds of biological (microbial) origin. The understanding of the physicochemical properties of biosurfactants such as surface tension reduction, dispersion, emulsifying, foaming or micelle formation is essential for the successful application of biosurfactants in many branches of industry. Glycolipids, which belong to the class of low molecular weight surfactants are currently gaining a lot of interest for industrial applications. For this reason, we focus mainly on this class of biosurfactants with particular emphasis on rhamnolipids and sophorolipids, the most studied of the glycolipids.

Toxicity Profiling of Biosurfactants Produced by Novel Marine Bacterial Strains.

Surface active agents (SAAs), currently used in modern industry, are synthetic chemicals produced from non-renewable sources, with potential toxic impacts on humans and the environment. Thus, there is an increased interest for the identification and utilization of natural derived SAAs. As such, the marine environment is considered a promising source of biosurfactants with low toxicity, environmental compatibility, and biodegradation compared to their synthetic counterparts. MARISURF is a Horizon 2020 EU-funded project aiming to identify and functionally characterize SAAs, derived from a unique marine bacterial collection, towards commercial exploitation. Specifically, rhamnolipids produced by Marinobacter MCTG107b and Pseudomonas MCTG214(3b1) strains were previously identified and characterized while currently their toxicity profile was assessed by utilizing well-established methodologies. Our results showed a lack of cytotoxicity in in vitro models of human skin and liver as indicated by alamar blue and propidium iodide assays. Additionally, the use of the single gel electrophoresis assay, under oxidative stress conditions, revealed absence of any significant mutagenic/anti-mutagenic potential. Finally, both 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonicacid) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) cell-free assays, revealed no significant anti-oxidant capacity for neither of the tested compounds. Consequently, the absence of significant cytotoxicity and/or mutagenicity justifies their commercial exploitation and potential development into industrial end-user applications as natural and environmentally friendly biosurfactants.

Congener-dependent conformations of isolated rhamnolipids at the vacuum-water interface: A molecular dynamics simulation.

HYPOTHESIS: Molecular dynamics simulation can be used to differentiate between the adsorption properties of rhamnolipid congeners at a vacuum-water interface. EXPERIMENTS: Adsorption of five congeners with differing alkyl chains (two C10 chains, two C14 chains or mixed C14C10 and C10C14), number of rhamnose rings (mono- or di-) and carboxyl group charge (non-ionic or anionic) are simulated at the vacuum-water interface. FINDINGS: All rhamnolipids adsorb in the interfacial region with rhamnose and carboxyl groups closer to the water phase, and alkyl chains closer to the vacuum phase, but with differing adsorbed conformations. Headgroups of uncharged congeners show two preferred conformations, closed and partially open. Di-rhamnolipid has a low proportion of closed conformation, due to the steric constraints of the second pyranose ring. Charged congeners show strong preference for closed headgroup conformations. For rhamnolipids with equal alkyl chains lengths (C10C10, C14C14) the distribution of alkyl chain tilt angles is similar for both. Where chain lengths are unequal (C14C10, C10C14) one chain has a greater tendency to tilt towards the water phase (>90°). The order parameter of the alkyl chains shows they are disordered at the interface. Together, these results show congener-dependent adsorbed conformation differences suggesting they will have differing surface-active properties at vacuum-water and oil-water interfaces.

Investigating the biomolecular interactions between model proteins and glycine betaine surfactant with reference to the stabilization of emulsions and antimicrobial properties.

Binding effect and interaction of 2-pentadecanoyloxymethyl)trimethylammonium bromide (DMGM-14) with bovine serum albumin (BSA) and hen egg white lysozyme (HEWL) were systematically investigated by the fluorescence spectroscopy, circular dichroism (CD) spectroscopy, isothermal titration calorimetry (ITC), surface tension analysis, and molecular docking studies. The emulsion properties and particle size distribution of surfactant/protein complexes containing sunflower oil were studied using static light scattering and confocal laser scanning microscopy (CLSM). The fluorescence spectroscopy and ITC analysis confirmed the complexes formation of DMGM-14 with BSA and HEWL which was also verified by surface tension measurements. CD results explained the conformational changes in BSA and HEWL upon DMGM-14 complexation. Molecular docking study provides insight into the binding of DMGM-14 into the specific sites of BSA and HEWL. Besides, the studies drew a detailed picture on the emulsification properties of DMGM-14 with BSA and HEWL. In addition, the in vitro experiment revealed a broad antibacterial spectrum of DMGM-14 and DMGM-14/HEWL complex including activity against Gram-positive and Gram-negative bacteria. In conclusion, the present study revealed that the interaction between DMGM-14 with BSA and HEWL is important for the pharmaceutical, biological, and food products.

Surface Active Agents and Their Health-Promoting Properties: Molecules of Multifunctional Significance.

Surface active agents (SAAs) are molecules with the capacity to adsorb to solid surfaces and/or fluid interfaces, a property that allows them to act as multifunctional ingredients (e.g., wetting and dispersion agents, emulsifiers, foaming and anti-foaming agents, lubricants, etc.) in a widerange of the consumer products of various industrial sectors (e.g., pharmaceuticals, cosmetics, personal care, detergents, food, etc.). Given their widespread utilization, there is a continuously growing interest to explore their role in consumer products (relevant to promoting human health) and how such information can be utilized in order to synthesize better chemical derivatives. In this review article, weaimed to provide updated information on synthetic and biological (biosurfactants) SAAs and their health-promoting properties (e.g., anti-microbial, anti-oxidant, anti-viral, anti-inflammatory, anti-cancer and anti-aging) in an attempt to better define some of the underlying mechanism(s) by which they exert such properties.

Marine-Derived Surface Active Agents: Health-Promoting Properties and Blue Biotechnology-Based Applications.

Surface active agents are characterized for their capacity to adsorb to fluid and solid-water interfaces. They can be classified as surfactants and emulsifiers based on their molecular weight (MW) and properties. Over the years, the chemical surfactant industry has been rapidly increasing to meet consumer demands. Consequently, such a boost has led to the search for more sustainable and biodegradable alternatives, as chemical surfactants are non-biodegradable, thus causing an adverse effect on the environment. To these ends, many microbial and/or marine-derived molecules have been shown to possess various biological properties that could allow manufacturers to make additional health-promoting claims for their products. Our aim, in this review article, is to provide up to date information of critical health-promoting properties of these molecules and their use in blue-based biotechnology (i.e., biotechnology using aquatic organisms) with a focus on food, cosmetic and pharmaceutical/biomedical applications.

Biosynthesis of rhamnolipid by a Marinobacter species expands the paradigm of biosurfactant synthesis to a new genus of the marine microflora.

BACKGROUND: In comparison to synthetically derived surfactants, biosurfactants produced from microbial culture are generally regarded by industry as being more sustainable and possess lower toxicity. One major class of biosurfactants are rhamnolipids primarily produced by Pseudomonas aeruginosa. Due to its pathogenicity rhamnolipid synthesis by this species is viewed as being commercially nonviable, as such there is a significant focus to identify alternative producers of rhamnolipids. RESULTS: To achieve this, we phenotypically screened marine bacteria for biosurfactant production resulting in the identification of rhamnolipid biosynthesis in a species belonging to the Marinobacter genus. Preliminary screening showed the strain to reduce surface tension of cell-free supernatant to 31.0 mN m-1. A full-factorial design was carried out to assess the effects of pH and sea salt concentration for optimising biosurfactant production. When cultured in optimised media Marinobacter sp. MCTG107b produced 740 ± 28.3 mg L-1 of biosurfactant after 96 h of growth. Characterisation of this biosurfactant using both HPLC-MS and tandem MS showed it to be a mixture of different rhamnolipids, with di-rhamnolipid, Rha-Rha-C10-C10 being the most predominant congener. The strain exhibited no pathogenicity when tested using the Galleria mellonella infection model. CONCLUSIONS: This study expands the paradigm of rhamnolipid biosynthesis to a new genus of bacterium from the marine environment. Rhamnolipids produced from Marinobacter have prospects for industrial application due to their potential to be synthesised from cheap, renewable feed stocks and significantly reduced pathogenicity compared to P. aeruginosa strains.

Hydrocarbon-degradation and MOS-formation capabilities of the dominant bacteria enriched in sea surface oil slicks during the Deepwater Horizon oil spill.

A distinctive feature of the Deepwater Horizon (DwH) oil spill was the formation of significant quantities of marine oil snow (MOS), for which the mechanism(s) underlying its formation remain unresolved. Here, we show that Alteromonas strain TK-46(2), Pseudoalteromonas strain TK-105 and Cycloclasticus TK-8 - organisms that became enriched in sea surface oil slicks during the spill - contributed to the formation of MOS and/or dispersion of the oil. In roller-bottle incubations, Alteromonas cells and their produced EPS yielded MOS, whereas Pseudoalteromonas and Cycloclasticus did not. Interestingly, the Cycloclasticus strain was able to degrade n-alkanes concomitantly with aromatics within the complex oil mixture, which is atypical for members of this genus. Our findings, for the first time, provide direct evidence on the hydrocarbon-degrading capabilities for these bacteria enriched during the DwH spill, and that bacterial cells of certain species and their produced EPS played a direct role in MOS formation.

Bacterial properties changing under Triton X-100 presence in the diesel oil biodegradation systems: from surface and cellular changes to mono- and dioxygenases activities.

Triton X-100, as one of the most popular surfactants used in bioremediation techniques, has been reported as an effective agent enhancing the biodegradation of hydrocarbons. However efficient, the surfactant's role in different processes that together enable the satisfying biodegradation should be thoroughly analysed and verified. In this research, we present the interactions of Triton X-100 with the bacterial surfaces (hydrophobicity and zeta potential), its influence on the enzymatic properties (considering mono- and dioxygenases) and profiles of fatty acids, which then all together were compared with the biodegradation rates. The addition of various concentrations of Triton X-100 to diesel oil system revealed different cell surface hydrophobicity (CSH) of the tested strains. The results demonstrated that for Pseudomonas stutzeri strain 9, higher diesel oil biodegradation was correlated with hydrophilic properties of the tested strain and lower Triton X-100 biodegradation. Furthermore, an increase of the branched fatty acids was observed for this strain.

Modification of surface and enzymatic properties of Achromobacter denitrificans and Stenotrophomonas maltophilia in association with diesel oil biodegradation enhanced with alkyl polyglucosides.

The article concerns the influence of selected alkyl polyglucosides on biodegradation, cell surface and enzymatic properties of Stenotrophomonas maltophilia and Achromobacter denitrificans. The biodegradation of diesel oil depends on several factors including type and the amount of surfactant as well as bacterial genera used in the process. Nevertheless, a careful selection of these variables must be made as some bacterial strains prefer to use surfactants as their carbon source. This leads to the lowered biodegradation of diesel oil as can be observed for the tested S. maltophilia strain. Alkyl polyglucosides influenced the cell surface properties of both of the tested strains in slightly different ways. Especially for A. denitrificans, for which the hydrophobicity increased with concentration of both--Lutensol GD 70 and Glucopon 215 in diesel oil-surfactant systems. Moreover, judging by the efficiency of biodegradation, the most effective process was observed in the presence of Lutensol GD 70 (240 and 360 mg L(-1)) with biodegradation rising from 32% (without surfactant) to 68%. No such relation was observed for S. maltophilia.

Influence of metal ions on bioremediation activity of protocatechuate 3,4-dioxygenase from Stenotrophomonas maltophilia KB2.

The aim of this paper was to describe the effect of various metal ions on the activity of protocatechuate 3,4-dioxygenase from Stenotrophomonas maltophilia KB2. We also compared activity of different dioxygenases isolated from this strain, in the presence of metal ions, after induction by various aromatic compounds. S. maltophilia KB2 degraded 13 mM 3,4-dihydroxybenzoate, 10 mM benzoic acid and 12 mM phenol within 24 h of incubation. In the presence of dihydroxybenzoate and benzoate, the activity of protocatechuate 3,4-dioxygenase and catechol 1,2-dioxygenase was observed. Although Fe(3+), Cu(2+), Zn(2+), Co(2+), Al(3+), Cd(2+), Ni(2+) and Mn(2+) ions caused 20-80 % inhibition of protocatechuate 3,4-dioxygenase activity, the above-mentioned metal ions (with the exception of Ni(2+)) inhibited catechol 1,2-dioxygenase to a lesser extent or even activate the enzyme. Retaining activity of at least one of three dioxygenases from strain KB2 in the presence of metal ions makes it an ideal bacterium for bioremediation of contaminated areas.

Cell surface properties and fatty acids composition of Stenotrophomonas maltophilia under the influence of hydrophobic compounds and surfactants.

Surface properties of newly isolated Stenotrophomonas maltophilia strain 6 were tested. The bacteria were stored in two different ways to determine the influence of hydrocarbons and surfactants on surface and enzymatic characteristics of the isolated strain. The influence of surface active agents, natural and synthetic, on membrane's lipid composition and cell surface hydrophobicity (CSH) was investigated. Our results indicate that long-term contact with diesel oil as a hydrophobic sole carbon source leads to the increased enzymatic activity of S. maltophilia strain 6 as well as to modification of fatty acids profiles and its facility to adhere to hydrophobic compounds. Among surfactants there were saponins and Triton X-100 which changed the composition of fatty acids the most, increasing the amount of branched acids. The comparison of fatty acid profiles with CSH of systems with diesel oil, rhamnolipids, saponins and Triton X-100 points out that the growing amount of hydroxy fatty acids corresponds to lower hydrophobicity. Moreover, CSH is a dynamic parameter which can change during cultivation of microorganisms.

Biodegradation of alkyl derivatives of aromatic hydrocarbons and cell surface properties of a strain of Pseudomonas stutzeri.

Pseudomonas stutzeri strain 9 was isolated from petroleum-contaminated soil. The main purpose of this study was to investigate how the long-term contact of this strain with diesel oil influences its surface and biodegradation properties. The experiments showed that the tested strain was able to degrade aromatic alkyl derivatives (butylbenzene, sec-butylbenzene, tert-butylbenzene and isobutylbenzene) and that the storage conditions had an influence on the cell surface properties. Also greater agglomeration of the cells was observed in the scanning electron microscope (SEM) micrographs and confirmed in particle size distribution results. The results also indicated that the addition of rhamnolipids to the hydrocarbons led to modification of the surface properties of P. stutzeri strain 9, which could be observed in the zeta potential and hydrophobicity values.