Extended Data Fig. 2: Purification and NMR analysis of enzymatically synthesized polymer.
From: A multi-enzyme machine polymerizes the Haemophilus influenzae type b capsule
a, Hib polymer (5 mg) was purified from an enzyme reaction containing RpiA, Bcs1, and Bcs3 using preparative AEC. The remaining nucleotide substrates and nucleotide products (see Fig. 1c) were UV-detectable at 280 nm, eluted early from the column and could be separated from b, the UV-inactive Hib polymer, which was visualized by Alcian blue / silver staining after PAGE. Polymer containing fractions were pooled, dialyzed against water, and c, analyzed by 1D and 2D NMR. All correlations are in agreement with published literature (Supplementary Table 1). Correlations confirming the linkage of the moieties in the repeating unit are highlighted in red in the structure and indicated by dashed lines. HMBC: heteronuclear multiple bond correlation, HSQC: heteronuclear single-quantum coherence.
