Rhamnolipids are naturally occurring biosurfactants with a wide range of potential commercial applications. As naturally derived products they present an ecological alternative to synthetic surfactants. The majority of described rhamnolipid productions are single strain Pseudomonas spp. cultivations. Here we report rhamnolipids producing bacteria Acinetobacter calcoaceticus, Enterobacter asburiae and Pseudomonas aeruginosa that were cultivated separately and as mixed populations. The ratio and composition of rhamnolipid congeners was determined by tandem mass spectrometry with negative electrospray ionization. Mono-rhamnolipid and di-rhamnolipid homologues containing one or two saturated or monounsaturated 3-hydroxy fatty acids were found in all strains. Physiochemical characterization of rhamnolipids was evaluated by the critical micelle concentration determination, the emulsification test, oil displacement test and phenanthrene solubilization. Critical micelle concentrations of rhamnolipids produced by both single strain and mixed cultures were found to be very low (10-63 mg/l) and to correspond with saturated/unsaturated fatty acid content of rhamnolipid homologues. The rhamnolipids produced by all strains effectively emulsified crude petroleum in comparison with synthetic surfactants Tween 80 and sodium dodecyl sulfate (SDS). Good performance of phenanthrene solubilization was exhibited by rhamnolipids from E. asburiae. The single strain and co-cultures cultivations were proposed as a possible way to produce rhamnolipid mixtures with a specific composition and different physiochemical properties, which could be exploited in bioremediation of various hydrophobic contaminants.
- MeSH
- Acinetobacter calcoaceticus metabolismus MeSH
- Enterobacter metabolismus MeSH
- fenantreny chemie metabolismus MeSH
- glykolipidy chemie metabolismus MeSH
- hmotnostní spektrometrie s elektrosprejovou ionizací MeSH
- povrchově aktivní látky chemie metabolismus MeSH
- průmyslová mikrobiologie MeSH
- Pseudomonas aeruginosa metabolismus MeSH
- rozpustnost MeSH
- tandemová hmotnostní spektrometrie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The possibility of utilizing volatile fatty acids (VFA)-containing waste substrates from biotechnological and industrial processes was investigated by cultivating both oleaginous (Candida sp., Rhodotorula glutinis, Trichosporon cutaneum, Yarrowia lipolytica) and non-oleaginous (Kluyveromyces polysporus, Saccharomyces cerevisiae, Torulaspora delbrueckii) yeast species on acetic acid, propionic acid and a combination of either acid with glucose as carbon and energy sources. Both oleaginous and non-oleaginous yeasts grew on VFA. Oleaginous yeasts accumulated lipids to 15-48% of dry cell weight, non-oleaginous yeasts also grew on VFA and showed comparable biomass yields but the lipid content was only 2-5%. Biomass and lipid yield increased in cultivations on VFA plus glucose. The lipid composition was comparable to plant-derived oils and therefore might be exploitable in biodiesel production; nearly all species, when cultured on propionate, showed a high content of the desirable odd-chain unsaturated FA, especially 17:1 acid. This study points at the wide array of possible applications of many yeasts, even non-oleaginous strains, for biovalorization of industrial wastes. Despite their low lipid content these species are useful because they can readily utilize VFA from waste products and, since they are not biologically hazardous, their biomass can be afterwards used, e.g. as livestock fodder.
Palmitoleic acid is found in certain dairy products and has broad applications in medicine and cosmetics. We tried to find a suitable producer of this acid among traditional biotechnological yeast species (Kluyveromyces polysporus, Torulaspora delbrueckii, Saccharomyces cerevisiae) characterized by high biomass yield and Candida krusei, Yarrowia lipolytica and Trichosporon cutaneum accumulating large amounts of lipids. The main factor affecting the content of palmitoleic acid was found to be the C/N ratio in the culture medium, with ammonium sulfate as an optimum nitrogen source leading to highest biomass yield with concomitantly increased lipid accumulation, and an increased content of ω6-linoleic acid, the precursor of prostaglandins, leukotrienes, and thromboxanes. We found that C. krusei can be conveniently used for the purpose, albeit only under certain cultivation conditions, whereas S. cerevisiae can produce high and stable amounts of palmitoleic acid in a broad range of cultivation conditions ranging from conventional to nutrient limitations.
- MeSH
- bioreaktory mikrobiologie MeSH
- druhová specificita MeSH
- kultivační média chemie metabolismus MeSH
- kvasinky klasifikace metabolismus MeSH
- kyseliny mastné mononenasycené izolace a purifikace metabolismus MeSH
- metabolismus lipidů fyziologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
The unflagging interest in the surfactants of biological origin, representing ecological alternatives to their synthetic counterparts, has enhanced R&D effort both to produce their new types and to resolve the bottlenecks of their commercialization. In this context, the rhamnolipids, offering a relatively large scale of potential applications, variety of congeners, low toxicity as well as stability towards the extremes of environment, logically attract attention. In this connection, the current state of knowledge concerning these compound exploitation, biosynthesis control and non-genetic factors affecting both production yield and final rhamnolipid product is surveyed.
Genus Pseudomonas includes a large number of species that can be encountered in biotechnological processes as well as in the role of serious human or plant pathogens. Pseudomonads easily form biofilms on various types of surfaces. The biofilm phenotype is characterized by an increased resistance to environmental influences including resistance to antibiotics and other disinfectants, causing a number of problems in health care, food industry, and other areas. Considerable attention is therefore paid to the possibilities of eradication/destruction of pseudomonads biofilms both in terms of understanding the mechanisms of biofilm formation and at the level of finding suitable antibiofilm tools applicable in practice. The first part of this review is devoted to an overview of the regulatory mechanisms that are directly or indirectly involved in the formation of biofilm. The most effective approaches to suppressing the formation of biofilm that do not cause the development of resistance are based on the application of substances that interfere with the regulatory molecules or block the appropriate regulatory mechanisms involved in biofilm development by the cells. Pseudomonads biofilm formation is, similar to other microorganisms, a sophisticated process with many regulatory elements. The suppression of this process therefore also requires multiple antibiofilm tools.
- MeSH
- antibakteriální látky farmakologie MeSH
- bakteriální léková rezistence MeSH
- biofilmy * MeSH
- fenotyp MeSH
- lidé MeSH
- nemoci rostlin mikrobiologie MeSH
- pseudomonádové infekce farmakoterapie mikrobiologie prevence a kontrola MeSH
- Pseudomonas účinky léků fyziologie MeSH
- půdní mikrobiologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
Rhamnolipid production by two non-pathogenic bacterial strains Acinetobacter calcoaceticus and Enterobacter asburiae, and established rhamnolipid producer Pseudomonas aeruginosa was investigated. Rhamnolipids were separated from supernatant and further purified by thin-layer chromatography. Mass spectrometry with negative electrospray ionization revealed rhamnolipid homologues varying in chain length and unsaturation. Tandem mass spectrometry identified mono-rhamnolipid and di-rhamnolipid homologues containing one or two 3-hydroxy fatty acids. Several media differing in carbon (sunflower oil, glycerol and sodium citrate), nitrogen (ammonium ions, nitrate) and phosphorus (total content) source, respectively, were tested to obtain enhanced rhamnolipid production. The best production (0.56g/l) was obtained when nitrate was used as a nitrogen source. Both strains produced rhamnolipids that exhibited excellent emulsification activity with aromatic and aliphatic hydrocarbons and several plant oils. Unlike P. aeruginosa the two strains, i.e. Acinetobacter and Enterobacter, are not pathogenic to humans.
- MeSH
- Acinetobacter calcoaceticus metabolismus MeSH
- Enterobacter metabolismus MeSH
- glykolipidy biosyntéza izolace a purifikace MeSH
- Pseudomonas aeruginosa metabolismus MeSH
- tandemová hmotnostní spektrometrie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
Bioremediation processes based on biofilms are usually very effective. The presence of (bio)surfactants in such processes can increase bioavailability of hydrophobic pollutants in aqueous phase. However, surfactants can affect the biofilm as well as individual microbial cells in different ways. Biosurfactants produced by a microbial population can be involved in the final structure of biofilm. An external application of synthetic surfactants or 'foreign' biosurfactants often results in partial or complete destruction of the biofilm and their high concentrations also have a toxic effect on microbial cells. Finding a suitable surfactant and its concentration, which would minimize the negative effects mentioned above, would allow to construct effective bioremediation processes using the benefits of both the biofilm and the surfactant. In this context, G(+) bacterium Rhodococcus erythropolis, which has a wide potential for biodegradation of aromatic compounds, was studied. High surface hydrophobicity of its cells, given mainly by the presence of mycolic acids in the cell envelopes, allows formation of stable biofilms. Three synthetic surfactants (Spolapon AOS 146, Novanik 0633A, Tween 80) and rhamnolipid isolated from Pseudomonas aeruginosa were used. Changes in initial adhesion and biofilm formation caused by the surfactants were monitored in a flow cell equipped with hydrophilic/hydrophobic carriers and analyzed by image analysis.
- MeSH
- aromatické uhlovodíky metabolismus MeSH
- biodegradace účinky léků MeSH
- biofilmy účinky léků MeSH
- fenoly metabolismus MeSH
- hydrofobní a hydrofilní interakce účinky léků MeSH
- látky znečišťující životní prostředí metabolismus MeSH
- micely MeSH
- počet mikrobiálních kolonií MeSH
- polyethylen MeSH
- povrchově aktivní látky farmakologie MeSH
- reologie účinky léků MeSH
- Rhodococcus účinky léků růst a vývoj fyziologie MeSH
- techniky vsádkové kultivace MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
We used reversed phase liquid chromatography-electrospray ionization tandem mass spectrometry for direct analysis of mycolic acids (MAs) from four different cultivations of Rhodococcus erythropolis. This technique enabled us to identify and quantify the specific molecular species of MAs directly from lipid extracts of the bacterium, including the determination of their basic characteristics such as retention time and mass spectra. We identified a total of 60 molecular species of MAs by means of LC/MS. In collision-induced dissociation tandem mass spectrometry, the [M-H](-) ions eliminated two residues, i.e., meroaldehyde and carboxylate anions containing α-alkyl chains. The structural information from these fragment ions affords structural assignment of the mycolic acids, including the lengths and number of double bond(s). Two strains, i.e., R. erythropolis CCM 2595 and genetically modified strain CCM 2595 pSRK 21 phe were cultivated on two different substrates (phenol and phenol with addition of humic acids as a sole carbon source). The addition of humic acids showed that there is a marked increase of unsaturated mycolic acids, mostly in the range of 20-100 %. This effect is more pronounced in the R. erythropolis CCM 2595 strain.
- MeSH
- biotransformace MeSH
- chromatografie kapalinová MeSH
- fenol metabolismus MeSH
- huminové látky MeSH
- kultivační média chemie MeSH
- kyseliny mykolové chemie metabolismus MeSH
- Rhodococcus chemie metabolismus MeSH
- tandemová hmotnostní spektrometrie MeSH
- uhlík metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Reversed phase liquid chromatography-atmospheric pressure chemical ionization mass spectrometry (RP-HPLC/MS-APCI) was used to identify and quantify triacylglycerols (TAGs) having odd-numbered ω-phenylalkanoic acids from seeds of the flower plant Dracunculus vulgaris, and TAGs from the bacterium Rhodococcus erythropolis prepared by precursor directed biosynthesis from phenylalanine and having the corresponding even-numbered ω-phenylalkanoic acids. Model compounds, which are not commercially available, were prepared by organic synthesis and this allowed us to extend the number of identified natural TAGs to nearly 140 molecular species. Both synthetic and natural compounds containing ω-phenylalkanoic acids were found to have antioxidant and free radical scavenging properties.
- MeSH
- Araceae chemie MeSH
- chromatografie s reverzní fází MeSH
- hmotnostní spektrometrie MeSH
- mastné kyseliny analýza chemie izolace a purifikace MeSH
- Rhodococcus chemie MeSH
- scavengery volných radikálů analýza chemie izolace a purifikace MeSH
- semena rostlinná chemie MeSH
- techniky syntetické chemie MeSH
- triglyceridy analýza chemie izolace a purifikace MeSH
- vysokoúčinná kapalinová chromatografie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
A biosynthetic pathway using pivalic acid as a starter unit was found in three bacterial species, Alicyclobacillus acidoterrestris, Rhodococcus erythropolis and Streptomyces avermitilis. When deuterium-labelled pivalic acid was added to A. acidoterrestris and R. erythropolis nutrient media it was incorporated into fatty acids to give rise to tert-butyl fatty acids (t-FAs). In addition, in R. erythropolis, pivalic acid was transformed into two starter units, i.e. isobutyric and 2-methylbutyric acid, which served as precursors of corresponding iso-even FAs and anteiso-FAs. In S. avermitilis the biosynthesis also yielded all three branched FAs; apart from this pathway, both pivalic and 2-methylbutyric acids were incorporated into the antibiotic avermectin.
- MeSH
- Alicyclobacillus metabolismus MeSH
- antibakteriální látky biosyntéza MeSH
- butyráty metabolismus MeSH
- ivermektin analogy a deriváty metabolismus MeSH
- kyseliny pentanové metabolismus MeSH
- mastné kyseliny biosyntéza MeSH
- Rhodococcus metabolismus MeSH
- Streptomyces metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
