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. Author manuscript; available in PMC: 2019 Dec 12.
Published in final edited form as: Biochemistry. 2017 Jan 17;56(4):612–622. doi: 10.1021/acs.biochem.6b00774

Figure 2.

Figure 2.

(a) 13C-CPMAS spectra of whole cells of E. faecalis grown in ESM supplemented with sodium-[2-13C]pyruvate (0.1 g/L). Vancomycin was added to final concentration of 10 μg/mL to one aliquot, and cells were grown for 1 hour and harvested. The 206-ppm peak assigned to [2-13C]pyruvate is absent from the 13C-CPMAS spectrum of VRE grown without vancomycin, due to [2-13C]pyruvate being fully utilized for the biosynthesis of [2-13C]Ala visible at 55 ppm (blue circle) and MurNAc at 72 ppm, which overlaps with the natural abundance 13C from glycans in PG. 13C-CPMAS spectra of VRE grown with and without vancomycin are results of 40000 and 80000 accumulated scans respectively. The magic angle spinning was at 5000 Hz. (b) The difference spectrum was obtained by subtracting 13C-CPMAS spectrum of VRE grown without vancomycin from the spectrum of VRE grown with vancomycin, and it shows increased d-[2-13C]Lac biosynthesis in VRE consistent with VanHB activity.