Heating block

Hydrolysis experiments without dialysis were carried out as follows. In general, a 200 μL aliquot of a 100 μM MINP stock solution in Millipore water was diluted by water or a 10 mM MES buffer (pH 6.0) to 990 μL and sonicated for 0.5 min. To this solution, a 10 μL aliquot of a 10/20 mM oligosaccharide stock solution was added. The reaction mixture was allowed to react in a Benchmark heating block at 60 or 90 °C for the indicated time. The reaction mixture was centrifuged (20 000 RPM for 10 min) to remove the MINP catalyst before LC-MS analysis using calibration curves generated from authentic samples (Fig. S32†). Hydrolysis experiments with dialysis were carried out as follows. In general, a 200 μL aliquot of a 100 μM MINP catalyst in Millipore water was diluted with Millipore water to 990 μL and sonicated for 0.5 min, and then the solution was added to a dialysis tubing (MWCO 500), followed by the addition of a 10 μL aliquot of a 10 mM maltohexaose stock solution (or 1 mg of amylose). The reaction mixture was dialyzed against 40 mL of Millipore water at 60 °C. The hydrolysis was monitored by LC-MS analysis of the external solution using calibration curves generated from authentic samples (Fig. S32†).