Effect of temperature on aminoglycoside binding sites in Stenotrophomonas maltophilia

In this study we used strains of Stenotrophomonas maltophilia grown at 30 degrees C and 37 degrees C to investigate the role of lipopolysaccharide (LPS) in temperature-dependent variations in sensitivity (TDVS) to gentamicin. TDVS was scored as 'good' if a four-fold or greater difference in minimum inhibitory concentration (MIC) was found between the two incubation temperatures (good TDVS strains; n = 23), and otherwise as 'poor' (poor TDVS strains; n = 15). Phosphate content of isolated LPS in the strains exhibiting good TDVS grown at 37 degrees C was significantly (P < 0.001) higher than those grown at 30 degrees C. However, the phosphate content from LPS of strains exhibiting poor TDVS did not alter significantly with growth temperature. There was no significant difference in 3-deoxy-D-manno-octulosonic acid (KDO) content between the strains grown at the different incubation temperatures. Fluorescence-activated cell sorting analysis showed significant differences in binding of fluorescein isothiocyanate conjugated gentamicin to cells grown at 30 degrees C or 37 degrees C. We conclude that the temperature-dependent variation in the aminoglycoside susceptibility of this species was not correlated with any detectable change in KDO content, but correlated well with phosphate content of LPS and that LPS phosphate is the major site of ionic interaction for aminoglycosides in S. maltophilia.