Analysis of the enzymatic activity of deglycosylated DNase I and endo-1, 4-β-mannosidase obtained from co- and post-fermentation with Endo H-P.

<p>(A). M DNA molecular weight markers (the size of each band was indicated on the left); Lane 1 pHBM905A plasmid (about 300 ng); Lane 2 pHBM905A treated with 1 μl supernatant of DNase I; Lane 3–4 pHBM905A treated with 0.5 and 1 μl of deglycosylated DNase I with post-fermentation treatment; Lane 5 pHBM905A treated with 1 U commercial DNase I; Lane 6–7 pHBM905A treated with 0.5 and 1 μl of deglycosylated DNase I with co-fermentation treatment; Lane 8 pHBM905A treated with fermentation supernatant from <i>P</i>. <i>pastoris</i> bearing pHM905A plasmid (the negative control). (B). A hole of about 2mm was made with a hole puncher on a MD plate supplemented with 1% konjac powder and 0.05% trypan blue. The samples were added into the wells and the plate was incubated at 37°C, overnight. Sample 1: 1.5 μl supernatant of <i>P</i>. <i>pastoris</i> expressing glycosylated endo-1, 4-β-mannosidase; Sample 2: 2 μl supernatant of <i>P</i>. <i>pastoris</i> expressing glycosylated mannanase; Sample 3: 2 μl supernatant of deglycosylated endo-1, 4-β-mannosidase with post-fermentation treatment; Sample 4: 2 μl supernatant of deglycosylated endo-1, 4-β-mannosidase with co-fermentation treatment; Sample 5: 2 μl supernatant of EndoH-P.