Chemicals | Relative activity (%) |
|---|
Metal ions (1 mM) |
None | 100 |
Ca2+
| 92.94 ± 1.65 |
Na+
| 93.29 ± 0.57 |
Mg2+
| 95.56 ± 1.01 |
Cu2+
| 43.77 ± 1.34 |
Fe3+
| 31 ± 0.38 |
Mn2+
| 162.54 ± 2.85 |
K+
| 97.35 ± 0.93 |
Other reagents (2 mM) |
SDS | 94.20 ± 2.04 |
EDTA | 86.88 ± 1.67 |
DTT | 134.67 ± 1.13 |
Urea | 100.42 ± 0.62 |
- The reaction was conducted in 20 mM Tris/HCl (pH 8) at 45 °C with 0.2 % (w/v) agarose. Metal ions (CaCl2, MgCl2, CuSO4, FeCl3, MnCl2, NaCl and KCl) were added to the reaction mixture to a final concentration of 1 mM in the absence of 1 mM EDTA. While, the inhibitors and denaturing reagents (EDTA, SDS, DTT, and urea) were added to the reaction mixture to a final concentration of 2 mM. Various types of reagents were measured under control blank samples. Relative activity was defined as the proportion of agarase activity for each treatment relative to agarase activity with no reagent. The agarase activity without metal and EDTA was taken to be 100 % (0.93 U ml−1). Relative activity (%): average relative activity ± SD. After preincubation with the chemicals for 10 min, the enzyme assay was performed
