burgdorferi can efficiently transport and utilize chitobiose in the absence of free GlcNAc to grow to optimal cell densities in one exponential phase, with optimal

growth occurring at chitobiose concentrations ≥ 18 μM. We confirmed those observations and also demonstrated that B. burgdorferi exhibits biphasic growth when cultured with low concentrations (≤ 15 μM) of chitobiose this website (Fig. 4A). This observation suggests that free chitobiose, and potentially longer free GlcNAc oligomers, are not the source of GlcNAc for growth in the second exponential phase, as was previously suggested [10]. In fact, growth of the wild type without GlcNAc but supplemented with longer GlcNAc oligomers, chitotriose and chitohexose, results in optimal cell densities and only one exponential phase (Rhodes and Nelson, manuscript in preparation).

This observation suggests that B. burgdorferi employs one or more enzymes for the breakdown of longer GlcNAc oligomers, and that this mechanism of obtaining sequestered (or MK0683 mw bound) GlcNAc in the form of chitin is turned on during the first exponential phase. Chitin and chitobiose may serve as an important nutrient source during the tick molt, as the peritrophic membrane encasing the blood meal is turned over and GlcNAc oligomers are released [8]. Previous laboratory studies by Tilly et al [11] demonstrated that chbC is not necessary for B. burgdorferi to complete an infectious cycle, leading them to suggest that the genome is still evolving and retains non-essential functional genes. However, we argue that selective pressure must be involved in the retention of this three component PTS, MycoClean Mycoplasma Removal Kit as it is also found in other Borrelia species (garinii and afzelli) that cause

Lyme borreliosis and to our knowledge there has not been a strain isolated in which this transport system is not present. This may be an instance in which mixed infection studies would be appropriate to determine the competitive index (i.e. degree of virulence attenuation) for the chbC mutant as compared to the wild type. To further demonstrate that free chitobiose or longer GlcNAc oligomers are not the source of GlcNAc in the second exponential phase, we followed the growth of cells in a medium lacking free GlcNAc and yeastolate (Fig. 8). Yeastolate is the only component of BSK-II that may contain GlcNAc oligomers, as it is derived from an organism with a chitinous cell wall. Tilly et al [10] previously reported that there was no second exponential phase by 250 hours when cells were cultured without free GlcNAc and yeastolate, and therefore, suggested that chitobiose and possibly other GlcNAc oligomers present in yeastolate may be the source of GlcNAc for growth in the second exponential phase. However, our results demonstrate that wild-type cells do exhibit a second exponential phase in the absence of free GlcNAc and yeastolate, and reach a peak cell density in the second exponential phase by 434 hours.