TABLE 4.
Sequential and simultaneous hydrolysis of CMC by CelZ and CelY
| Enzyme (relative proportion)a | Measured reducing sugar released (μmol/ml)b | Predicted activity from the arithmetic sum of CelY and CelZ (μmol/ml)c | Synergybd |
|---|---|---|---|
| CelZ (10 parts) + CelY (0 parts) | 4.65 ± 0.08 | 4.65 | 1.00 ± 0.02 |
| CelZ (0 parts) + CelY (10 parts) | 4.14 ± 0.04 | 4.14 | 1.00 ± 0.01 |
| CelZ (9 parts) + CelY (1 part) (simultaneously) | 8.28 ± 0.08 | 4.60 | 1.80 ± 0.02 |
| CelZ (9 parts) and then CelY (1 part) (sequentially) | 4.86 ± 0.23 | 4.60 | 1.06 ± 0.05 |
| CelY (1 part) and then CelZ (9 parts) (sequentially) | 8.75 ± 0.14 | 4.60 | 1.90 ± 0.03 |
a
CelZ and CelY were diluted to equal CMCase activities. Both simultaneous and sequential hydrolysis reactions (0.15 IU/ml) were investigated using 9 parts CelZ and 1 part CelY. In the sequential hydrolysis experiments, the first enzyme was incubated with the substrate for 4 h and inactivated by boiling for 20 min. After cooling, the second enzyme was added and incubated for an additional 4 h. All reactions were terminated by boiling.
b
Averages ± standard deviations (three experiments).
c
Calculated sum of individual CelY and CelZ activities.
d
Synergy was calculated as the observed activity divided by the sum of predicted contributions from CelY alone (10%) plus CelZ alone (90%).