Fig. 1.

(A) HPTLC analysis of mono-O-acetylated Neu5,9Ac₂ and (B) Neu5,4 Ac₂ incubated with NanS-p EHEC O-acetylesterases. Three μl of 1 mg/mL Neu5Ac (Fig. 1A,B, lanes 1,8), Neu5,9Ac₂ (Fig. 1A, lanes 2,9) and Neu5,4 Ac₂ (Fig. 1B, lanes 2,9) preparations each, were applied onto HPTLC plates as controls. In addition, three μl of 1 mg/ml Neu5,9Ac₂ (Fig. 1A, lane 3), and Neu5,4 Ac₂ (Fig. 1B, lane 3) containing reaction buffer without enzymes were applied as buffer controls. Neu5,9Ac₂ was digested with NanS-His (Fig.1A, lane 4), NanS-p1-His (Fig.1A, lane 5), NanS-p2-His (Fig.1A, lane 6) and NanS-p4-His (Fig.1A, lane 7). Neu5,4 Ac₂ was digested with NanS-His (Fig.1B, lane 4), NanS-p1-His (Fig. 1B, lane 5), NanS-p2-His (Fig. 1B, lane 6), and NanS-p4-His (Fig. 1B, lane 7).