Peptidoglycan and Teichoic Acid Levels and Alterations in <i>Staphylococcus aureus</i> by Cell-Wall and Whole-Cell Nuclear Magnetic Resonance

Gram-positive bacteria surround themselves with a multilayered macromolecular cell wall that is essential to cell survival and serves as a major target for antibiotics. The cell wall of <i>Staphylococcus aureus</i> is composed of two major structural components, peptidoglycan (PG) and wall teichoic acid (WTA), together creating a heterogeneous and insoluble matrix that poses a challenge to quantitative compositional analysis. Here, we present ¹³C cross polarization magic angle spinning solid-state nuclear magnetic resonance (NMR) spectra of intact cell walls, purified PG, and purified WTA. The spectra reveal the clear molecular differences in the two polymers and enable quantification of PG and WTA in isolated cell walls, an attractive alternative to estimating teichoic acid content from a phosphate analysis of completely pyrolyzed cell walls. Furthermore, we discovered that unique PG and WTA spectral signatures could be identified in whole-cell NMR spectra and used to compare PG and WTA levels among intact bacterial cell samples. The distinguishing whole-cell ¹³C NMR contributions associated with PG include the GlcNAc-MurNAc sugar carbons and glycyl α-carbons. WTA contributes carbons from the phosphoribitol backbone. Distinguishing ¹⁵N spectral signatures include glycyl amide nitrogens in PG and the esterified d-alanyl amine nitrogens in WTA. ¹³C NMR analysis was performed with samples at natural abundance and included 10 whole-cell sample comparisons. Changes consistent with altered PG and WTA content were detected in whole-cell spectra of bacteria harvested at different growth times and in cells treated with tunicamycin. This use of whole-cell NMR provides quantitative parameters of composition in the context of whole-cell activity

Mechanochemical Synthesis of Elusive Fluorinated Polyacetylene

Polymer mechanochemistry has traditionally been employed to study the effects of mechanical force on one or two chemical bonds within a polymer. It is underexploited for the scalable synthesis of wholly new materials by activating bonds along the entire polymer, especially products inaccessible by other means. Herein we utilize polymer mechanochemistry to synthesize fluorinated polyacetylene, a long-sought-after air-stable polyacetylene that has eluded synthesis by conventional means. Our synthetic approach proceeds via ultrasonication of a force-responsive precursor polymer that was synthesized in five steps on gram scale. The synthesis is highlighted by rapid incorporation of fluorine in an exotic photochemical cascade whose mechanism and exquisite diastereoselectivity were elucidated by computation. </p