An alternative infrared spectroscopy assay for the quantification of polysaccharides in bacterial samples.
The ability of bacteria to produce extracellular polysaccharides has been regarded as an indication of biofilm-forming capacity. Therefore, the determination of the sugar content in bacterial samples becomes a significant parameter. The colorimetric methods currently used are rather sensitive to the nature of the sugars and therefore require knowledge of the sugar types present in the samples. Unfortunately, the types of sugars present in bacteria are generally unknown and often composed of a complex mixture. In this article, we propose an alternative method based on Fourier transform infrared (FTIR) spectroscopy for the estimation of the total sugar content in bacterial samples. The method is based on a systematic treatment of FTIR spectra obtained from dried bacteria samples. It is assumed that the total sugar amount can be estimated from the area of characteristic bands between 970 and 1182 cm(-1). In parallel, the amide II band (1560-1530 cm(-1)) associated with proteins, or the C-H stretching region (2820-3020 cm(-1)) associated with the biomass, can be used for normalization purposes. Therefore, the ratio of the band area in the sugar window over that of the amide II or C-H stretching can be used to report the sugar content in bacterial samples. This method has been validated on model bacterial mixtures containing sugars, proteins, and DNA. Results with real bacterial samples are also provided and show conclusively that increased sugar contents in biofilms can be identified. The proposed FTIR approach requires minimal sample preparation and a single acquisition, is rapid, and may be applied to any kind of bacterial growth.
Marcotte
Lucie
L
Department of Chemistry, University of Montreal, Montreal, Que., Canada H3C 3J7.
Kegelaer
Grégory
G
Sandt
Christophe
C
Barbeau
Jean
J
Lafleur
Michel
M
D000818:Animals; D001419:Bacteria / Q000737:chemistry*; D018441:Biofilms; D002417:Cattle; D003124:Colorimetry / Q000379:methods; D004247:DNA / Q000032:analysis; D006801:Humans; D008954:Models, Biological; D011135:Polysaccharides, Bacterial / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D011550:Pseudomonas aeruginosa / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods*; D013212:Staphylococcus epidermidis / Q000737:chemistry; D013295:Streptococcus mutans / Q000737:chemistry; D016959:Xanthomonas campestris
D011135:Polysaccharides, Bacterial; D004247:DNA; C002563:xanthan gum