TABLE 5.
Formation of maltose-1-P from glycogen by mycobacterial enzyme fraction
Each incubation mixture contained 25,000 cpm of [14C]glycogen, Tris buffer, and enzyme (25 μg of protein) from the DE52 column. Various amounts of inorganic phosphate or arsenate were added, and incubations were for 30 min. The amount of radioactivity binding to DE52 (i.e. sugar-P) was determined. The DE52 eluted radioactivity was treated with alkaline phosphatase to remove covalently bound phosphate, and the neutralized sugars were subjected to paper chromatography as shown in Fig. 6.
| Additions to incubation | Radioactivity bound to DE52 |
|---|---|
| None | 460 |
| 0.5 mmol of Pi | 2100 |
| 1 mmol of Pi | 6600 |
| 5 mmol of Pi | 6500 |
| 2.5 mmol of A5O4 | 480 |
| Heat-killed enzyme + 2.5 mmol of Pi | 610 |