Abstract:
Microbial glycolipids consist of four major groups, rhamnolipids, sophorolipids, trehalose lipids and mannosylerythritol lipids. Extensive research has been carried out on rhamnolipids and sophorolipids, with slightly less research to date carried out on trehalose lipids and mannosylerythritol lipids. When studying these microbial glycolipids, the ability to isolate, purify and characterize the structures being produced is extremely important. This structural information provides insight into the different conditions, such as carbon sources, etc. that effect production of glycolipids. The information from analysis allows the optimization of production yields and assembly of glycolipids with different structural characteristic. Therefore, the ability to drive production in a certain direction allow the microbiologist to produce different types of glycolipids depending on the biological activity required, such as surface tension, is possible. The experimental techniques used to isolate, purify and analysis glycolipids is extremely varied, such as colorimetric assays that give rough indication of production yields, ranging to complex mass spectral techniques. Mass spectrometry provides essential information that results in the identification and quantification of individual glycolipid structures, including isomers. However, mass spectrometry requires extremely purified glycolipids for best result, which can be carried out using various chromatographic techniques. This paper therefore details information and methods that would be required to analysis glycolipids. Since there are numerous methods available, only the most commonly reported techniques are presented in this paper.
Keywords
- Nuclear Magnetic Resonance
- Thin Layer Chromatography
- Matrix Assisted Laser Desorption Ionization
- Evaporative Light Scatter Detector
- Elution Condition
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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- 1.
- 2.
When larger volumes need to be extracted, autoclaving to sterilise without removal of cells may be an advantage.
- 3.
Chloroform:methanol (2:1, v/v) can also be used for extraction, however the time taken for the two layers to separate is much greater.
- 4.
MTBE has also been shown to be a suitable solvent for extraction of trehalose lipids (Kuyukina et al., 2001).
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Use a glycolipid standard appropriate to the biosurfactant under investigation.
- 6.
Depending on how much separation is required, increments can range from 2% increases to 10% increases in methanol concentration.
- 7.
As well as a variety of columns that can be used for analyses, the other conditions such as run temperatures and times can also be varied.
- 8.
Other columns and consequently different GC-MS conditions can be used.
- 9.
Other gradient conditions or slight modifications should be used to achieve appropriate separation depending on the column selected.
- 10.
It is possible to use other columns and change the elution conditions.
- 11.
It is possible to use other columns and change the elution conditions.
- 12.
Method shown refers to the LCQ with ESI-MS carrying Excalibur software; for different equipment or software consult the manufacturers’ manuals.
- 13.
Signal is severely influenced by the presence of salts and other impurities.
References
Asmer H-J, Lang S, Wagner F, Wray V (1988) Microbial production, structure elucidation and bioconversion of sophorose lipids. J Am Oil Chem Soc 65: 1460–1466.
Asselineau C, Asselineau J (1978) Trehalose-containing glycolipids. Prog Chem Fats Other Lipid 16: 59–99.
Banat IM, Makkar RS, Cameotra SS (2000) Potential commercial applications of microbial surfactants. Appl Microbiol Biotechnol 53: 495–508.
Brandl H, Gross RA, Lenz RW, Fuller RC (1988) Pseudomonas oleovorans as a source of poly (β-hydroxyalkanoates) for potential application as biodegradable polyesters. Appl Environ Microbial 54: 1977–1982.
Chandrasekaran EV, Bemiller JN (1980) In Whistler RL (ed) Methods in Carbohydrate Chemistry, Vol 9. New York: Academic Press, p. 89.
Cavalero DA, Cooper DG (2003) The effect of medium composition on the structure and physical state of sophorolipids produced by Candida bombicola ATCC 22214. J Biotechnol 103: 31–41.
Davilla AM, Marhcal R, Monin N, Vandecasteele JP (1993) Identification and determination of individual sophorolipids in fermentation products by gradient elution high-performance liquid chromatography with evaporative light-scattering detection. J Chromatogr A 648: 139–149.
De Koster CG, Vos B, Versluis C, Heerma W, Haverkamp J (1994) High-performance TLC/fast atom bombardment (tandem) mass spectrometry of Pseudomonas rhamnolipids. Biol Mass Spectrom 23: 179–185.
De Koster CG, Heerma W, Pepermans HAM, Hroenewegen A, Peters H, Haverkamp J (1995) Tandem mass spectrometry and nuclear magnetic resonance spectroscopy studies of Candida bombicola sophorolipids and product formed on hydrolysis by cutinase. Anal Biochem 230: 135–148.
Desai JD, Banat IM (1997) Microbial production of surfactants and their commercial potential. Microbiol Mol Biol Rev 61: 47–64.
Deziel E, Lepine F, Dennie D, Boismenu D, Mamer OA, Villemur R (1999) Liquid chromatography/mass spectrometry analysis of mixtures of rhamnolipids produced by Pseudomonas aeruginosa strain 57RP grown on mannitol or naphthalene. Biochim Biophys Acta 1440: 244–252.
Deziel E, Lepine F, Milot S, Villemur R (2000) Mass spectrometry monitoring of rhamnolipids from a growing culture of Pseudomonas aeruginosa strain 57RP. Biochim Biophys Acta 1485: 145–152.
Fukuoka T, Morita T, Konishi M, Imura T, Sakai H, Kitamoto D (2007) Structural characterisation and surface-active properties of a new glycolipid biosurfactant, mono-acylated mannosylerythritol lipid, produced from glucose by Pseudozyma antarctica. Appl Microbiol Biotechnol 76: 801–810.
Heyd M, Kohnert T-H T, Nusser M, Kirschhofer F, Brenner-Weiss G, Franzreb M, Berensmeier S (2007) Development and trends of biosurfactant analysis and purification using rhamnolipids as an example. Anal Bioanal Chem 391: 1579–1590.
Hodge JE, Hofreiter BT (1962) Determination of reducing sugars and carbohydrates. Methods Carbohydr Chem 1: 380–394.
Hu Y, Ju L-K (2001a) Purification of lactonic sophorolipids by crystallisation. J Biotechnol 87: 263–272.
Hu Y, Ju L-K (2001b) Sophorolipid production from different lipid precursors observed with LC-MS. Enzyme Microb Technol 29: 593–601.
Ishigami Y, Gama Y, Nagahora H, Yamaguchi M, Nakahara H, Kamata T (1987) The pH-sensitive conversion of molecular aggregates of rhamnolipid biosurfactant. Chem Lett 16: 763–766.
Itoh S, Honda H, Tomita F, Suzuki T (1971) Rhamnolipids produced by Pseudomonas aeruginosa grown on n-paraffin. J Antibiot 24: 855–859.
Kitamoto D, Akiba S, Hioki C, Tabuchi T (1990a) Production of mannosylerythritol lipids by Candida antarctica from vegetable oil. Agric Biol Chem 54: 37–40.
Kitamoto D, Akiba S, Hioki C, Tabuchi T (1990b) Extracellular accumulation of mannosylerthritol lipids by a strain of Candida antarctica. Agric Biol Chem 54: 31–36.
Koch AK, Kappeli O, Fiechter A, Reiser J (1991) Hydrocarbon assimilation and biosurfactant production in Pseudomonas aeruginosa mutants. J Bacteriol 173: 4212–4219.
Konishi M, Morita T, Fukuoha T, Imura T, Kakugawa K, Kitamoto D (2007) Production of different types of mannosylerythritol lipids as biosurfactant by the newly isolated yeast strains belonging to the genus Pseudozyma. Appl Microbiol Biotechnol 75: 521–531.
Kosaric N (1993) Surfactant science, vol 48. New York: Marcel Dekker, p. 483.
Kretschmer A, Bock H, Wagner F (1982) Chemical and physical characterisation of interfacial-active lipids from Rhodococcus erythropolis grown on n-alkanes. Appl Environ Microbiol 44: 864–870.
Kuyukina MS, Ivshina IB, Philp JC, Christofi N, Dunbar SA, Ritchkova MI (2001) Recovery of Rhodococcus biosurfactants using methyl tertiary-butyl ether extraction. J Microbiol Methods 46: 149–156.
Lang S, Philp JC (1998) Surface-active lipids in Rhodococci. Int J General Mol Microbiol 74: 59–70.
Mata-Sandoval JC, Karns J, Torrents A (1999) High-performance liquid chromatography of rhamnolipid mixtures produced by Pseudomonas aeruginosa UG2 on corn oil. J Chromatogr A 684: 211–220.
Monteiro SA, Sassaki GL, De Souza LM, Meira JA, De Araujo JM, Mitchell DA, Ramos LP, Krieger N (2007) Molecular and structural characterization of the biosurfactant produced by Pseudomonas aeruginosa DAUPE 614. Chem Phys Lipids 147: 1–13.
Noordman WH, Brusseau ML, Janssen DB (2000) Adsorption of a multicomponent rhamnolipid surfactant to soil. Environ Sci Technol 34: 832–838.
Nunez A, Ashby R, Foglia TA, Solaiman DKY (2001) Analysis and characterization of sophorolipids by liquid chromatography with atmospheric pressure chemical ionization. Chromatographia 53: 673–677.
Rapp P, Bock H, Wray V, Wagner F (1979) Formation, isolation and characterisation of trehalose dimycolates from Rhodococcus erythropolis grown on n-alkanes. J Gen Microbiol 115: 491–503.
Rau U, Nguyen LA, Schulz S, Wray V, Nimtz M, Roeper H, Koch H, Lang S (2005) Formation and anaylsis of mannosylerythritol lipids secreted by Pseudozyma aphidis. Appl Microbiol Biotechnol 66: 551–559.
Schenk T, Schuphan I, Schmidt B (1995) High-performance liquid chromatography determination of the rhamnolipids produced by Pseudomonas aeruginosa. J Chromatogr A 693: 7–13.
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Smyth, T.J.P., Perfumo, A., Marchant, R., Banat*, I.M. (2010). Isolation and Analysis of Low Molecular Weight Microbial Glycolipids. In: Timmis, K.N. (eds) Handbook of Hydrocarbon and Lipid Microbiology. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-540-77587-4_291
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DOI: https://doi.org/10.1007/978-3-540-77587-4_291
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