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. 2017 Jan 16;7:40478. doi: 10.1038/srep40478

Figure 7. Enzymatic activity of BtGH53 on GOS.

Figure 7

(a) TLC analysis of the products formed by enzymatic digestion of BtGH53 on GOS mixture. Lane 1: Pectic galactan products standard. Lane 2: GOS, Lane 3: GOS + BtGH53. 10 μl of BtGH53 was incubated with 10 μl of 1% GOS in reaction buffer for 16 h at 37 °C. Afterwards, 2 × 2 μl of each reaction was spotted on a TLC plate and run for 4 h in solvent system containing 3:1:1 isopropanol, ethyl acetate, water. Spots were visualized using stain containing 20% sulfuric acid, 80% methanol and 0.5% orcinol after heating at 110 °C for 20 min. (b) Corresponding HPAEC-PAD analysis of BtGH53 activity on GOS (dotted line: before incubation; solid line: after incubation) showing depletion of higher DP GOS molecules and accumulation of galactobiose. (A) DP3 Branched GOS (left to right) β-D-Galp-(1 → 4)-[β-D-Galp-(1 → 6)-]D-Glcp, β-D-Galp-(1 → 2)-[β-D-Galp-(1 → 4)-]D-Glcp and β-D-Galp-(1 → 2)-[β-D-Galp-(1 → 6)-]D-Glcp, β-D-Galp-(1 → 3)-[β-D-Galp-(1 → 6)-]D-Glcp. (B) compounds in (A) with an additional β-D-Galp-(1 → 4)-linked residue on the non-reducing end on either galactose residue.