FTIR spectroscopic characterization of Escherichia coli

Fourier-Transform Infrared (FT-IR) spectroscopy is a widely utilized technique for chemical analysis (Johnston, 1991). In soil microbial biomass, IR spectroscopy appeared useful in establishing the spectral fingerprints of different cell constituents, such as proteins, lipids, carbohydrates and nucleic acid (Filip, 1978a, b). Time-consuming and difficult isolation of individual cell structural units can be omitted using this technique, when intact cells (biomass) are under investigation. Also, small amounts of biomass (1-2 mg) are sufficient for the analysis, and the spectra can be easily stored in the data files of most instruments. Naumann et al. (1990, 1991) documented the usefulness of FT-IR spectroscopy in bacterial diagnostics, including that of numerous E. coli strains. This potentially pathogenic bacterium inhabits mammalian intestine, and thus, it is widely used as an indicator of faecal water pollution. In our investigations (Filip et al., in press), the cell mass of E. coli delivered well-differentiated IR-bands (see Table 1).

As shown in Fig. 4, the FT-IR spectra of E. coli changed rather markedly with the age in hours of the culture. The same was true for biomass harvested from differently composed nutrient media (not shown in figure). From these results one can conclude that FT-IR spectra, seem only capable of delivering useful and reproducible information on bacteria under testing if the cell mass is obtained under well standardized cultural conditions.

Figure 4. FT-IR spectra of E. coli cell mass harvested after (A) 3 h, (B) 16 h, (C) 41 h, and

(D) 168 h from a minimum nutrient broth (Filip et al., in press)

Figure 4. FT-IR spectra of E. coli cell mass harvested after (A) 3 h, (B) 16 h, (C) 41 h, and

(D) 168 h from a minimum nutrient broth (Filip et al., in press)

References

Claus, H., Rötlich, H. and Filip, Z., 1992: Survival in groundwater of some bacteria with natural and recombinant plasmids. Microbial Releases, 1: 103-110.

Coates, J.D. and Achenbach, L.A., 2002: The biogeochemistry of aquifer systems. In: Hurst, C.J. (Ed.), Manual of Environmental Microbiology, Washington, DC: ASM Press, pp. 719-727.

Filip, Z., 1978a: Infrared spectroscopy of two soils and their components. In: Krumbein, W. (Ed.), Environmental Biogeochemistry and Geomicrobiology, Ann Arbor, MI: Ann Arbor Sci. Publ., pp. 747-754.

Filip, Z., 1978b: Infrarotspektren der mikrobiellen Biomasse und der Huminsäure im Podzolboden. Zeitschrift für Pflanzenernährung und Bodenkunde, 141: 711-715.

Filip, Z., Hermann, S. and Demnerova, K., 2008: FT-IR spectroscopic characteristics of differently cultivater Escherichia coli. Czech J. Food Sci. 26: 458-463.

Filip, Z., Kaddu-Mulindwa, D. and Milde, G., 1988: Survival of some pathogenic and facultative pathogenic bacteria in groundwater. Water Science and Technology, 20: 227-231.

Johnston, S.F., 1991: Fourier Transform Infrared - A Constantly Evolving Technology. New York: Ellis Horwood.

Knorr, M., 1951: Zur hygienischen Beurteilung der Ergänzung und des Schutzes großer Grundwasserverokommen. Grundwasser f., 92: 104-110.

Naumann, D., 2000: Infrared spectroscopy in microbiology. In: Meyers, R.A. (Ed.), Encyclopedia of Analytical Chemistry. Chichester: Wiley, pp. 102-131.

Naumann, D., Helm, D. and Labischinski, H., 1990: Einsatzmöglichkeiten der FT-IR Spektroskopie in Diagnostik und Epidemiologie. Bundesgesundheitsblatt, 33: 387-393.

Naumann, D., Helm, D. and Labischinski, H., 1991: Microbiological characterizations by FT-IR spectroscopy. Nature, 351: 81-82.

Parker, F.S., 1971: Infrared Spectroscopy in Biochemistry, Biology and Medicine. London: Hilger.

Pelikan, V., 1983: Groundwater Protection. Praha: SNTL (in Czech).

Schleyer, R. and Kerndorff, H., 1992: Die Grundwasserqualität westdeutscher

Trinkwasser-ressourcen. Weinheim: VCH. Smith, R.L., 2002: Determining the terminal electron-accepting reactions in the saturated subsurface. In: Hurst, C.J. (Ed.), Manual of Environmental Microbiology, Washington, DC: ASM Press, pp. 743-752. Stetzenbach, L.D., Kelley, L.M. and Sinclair, N.A., 1986: Isolation, identification, and growth of well-water bacteria. Ground Water, 24: 6-10. Ward, C.H., Giger, W. and McCarty, P.L. (Eds.), 1985: Ground Water Quality. New York: Wiley.

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