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. 2012 Aug 31;8(8):1085–1096. doi: 10.7150/ijbs.4406

Figure 6.

Figure 6

HPLC analysis of hydrolyzed products of glycans by OfHex2. Pyridylaminated substrates (10 μM) were incubated with purified OfHex2 at pH 5.5, 30 °C. Recombinant OfHex1 which exhibites no hydrolytic activity toward GnGn-PA, Gb4-PA and GA2-PA was used as negative control (with same activity toward pNP-β-GlcNAc). For GnGn-PA degradation analysis, OfHex1 exhibited no hydrolytic activity after incubation for 8 h (A), while OfHex2 degraded GnGn-PA to GnM-PA, MGn-PA and MM-PA after incubated for 30 min (B), and continued to degrade the intermediates GnM-PA and MGn-PA to MM-PA after a 4h-incubation (C). For Gb4-PA degradation analysis, OfHex1 exhibited no hydrolytic activity after incubation for 8 h (D), while OfHex2 degraded Gb4-PA to pyridylaminated globotriose (Gb3-PA) after 2 h-incubation (E). For GA2-PA degradation analysis, OfHex1 exhibited no drolytic activity after incubation for 8 h (F), while OfHex2 degraded GA2-PA to lactose-PA after 2 h-incubation (G).