TABLE 3.
Effect of various chemicals on Smlt1743 lyase activity
Enzyme was incubated in 30 mm buffer containing NaCl, CaCl2, or Vcpal at the indicated concentrations for 10 min before adding 17.5 μg (for poly-ManA and HA) or 1.75 μg (for poly-GlcA) to 1 mg/ml substrate in 30 mm buffer with the equivalent concentration of chemical. Enzyme activity was monitored by change in absorbance at 235 nm. Activity in the absence of any chemicals was taken to be 100%.
| Chemical | Concentration | Relative activity |
||
|---|---|---|---|---|
| Poly-GlcA (pH 7) | Poly-ManA (pH 9) | HA (pH 5) | ||
| mm | % | |||
| None | 100 ± 0.7 | 100 ± 4.2 | 100 ± 3.2 | |
| NaCl | 10 | 79.9 ± 5.6 | 104.0 ± 2.0 | 75.6 ± 0.2 |
| 50 | 29.6 ± 1.4 | 26.5 ± 1.0 | 0.5 ± 0.2 | |
| 200 | 9.0 ± 0.5 | 3.2 ± 0.3 | NDa | |
| CaCl2 | 1 | 68.5 ± 0.6 | 70.7 ± 8.8 | 73.5 ± 3.0 |
| 2 | 32.9 ± 0.8 | 27.1 ± 4.9 | 58.9 ± 2.3 | |
| Vcpal | 0.05 | 59.1 ± 3.7 | 64.5 ± 1.5 | 72.7 ± 5.0 |
| 0.1 | 29.9 ± 2.1 | 31.4 ± 1.1 | 45.6 ± 3.1 | |
a No detectable activity (detection limit: 0.001 absorbance units at 235 nm per minute).