Skip to main content
. 2014 Apr 15;9(4):e95024. doi: 10.1371/journal.pone.0095024

Figure 2. Enzymatic transformation of Lac-Enk to Gal-Lac-Enk.

Figure 2

A) RP-HPLC analysis of one-pot reaction for Lac-Enk with UDP-Glc and galactose epimerase. From top to bottom: substrate (Lac-Enk) only; positive control – reaction mixture containing UDP-Gal and LgtC; negative control – reaction mixture containing UDP-Glc and LgtC; reaction mixture containing UDP-Glc, LgtC and galactose epimerase. All reactions were conducted in 50 mM HEPES (pH 7), 10 mM MnCl2 buffer at 37°C. RP-HPLC was conducted on an Alltima C18 column using a gradient of 0–90% solvent B, tR = 17.2 min (substrate, Lac-Enk) and tR = 16.8 min (product, Gal-Lac-Enk). B) ES-MS analysis of the acceptor, Lac-Enk m/z (C44H63N7O18, 977.4) m/z 978.5 [M+H]+ (calcd. 978.4). C) ES-MS analysis of the product, Gal-Lac-Enk (C52H76N8O23, 1139.5) m/z 1140.5 [M+H]+ (calcd. 1140.5).