Figure 4. Analysis of SAP(H+L) enzymatic activity on pullulan and glycogen.

(A) NMR spectra indicate the generation of maltotriose units after the addition of SAP(H+L) to the reaction mixture containing pullulan. Pullulan NMR spectra were recorded on the native polysaccharide (−SAP) and after the addition of the recombinant enzyme (+SAP). NMR experiments were recorded at 25°C on Bruker Avance 600 MHz spectrometer and using 5-mm probe (Bruker). For details see the Experimental Procedures section. (B) SEC-HPLC analysis indicates that SAP(H+L) is active on glycogen. Two chromatograms were recorded at 214nm, one on the native glycogen polysaccharide (black line) and the other 1 h later the addition of SAP(H+L) (blue line). A gel filtration analytical column with a fractionation range of Mw PEG/PEO 2×10³–3×10⁵ Da was used. For details see the Experimental Procedures section.