20*25
PJSRV05-4
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Control experiment of recombinant enterokinase digestion at 4°C, 25°C, and 37°C temperatures.
Recombinant enterokinase test batch: REK160401. The substrate was digested according to the following method, and electrophoresis was used to confirm the results after certain time intervals. A total of 0.1 U of recombinant enterokinase was used to digest 50 µg of substrate for analysis.
| Time / h | 8 | 16 | 24 | 40 | 48 | 64 |
| Temperature | 4–8°C and 25°C respectively | |||||
| Time / h | 2 | 4 | 6 | 8 | 24 | |
| Temperature | 37°C |
The digestion at 4°C showed the following results:
RESULTS: The substrate was successfully digested at 4°C, but the process required an extended incubation time of 48–64 hours. This suggests that the enzyme activity is significantly lower at this temperature. The digestion at 25°C produced the following outcomes:
RESULTS: At 25°C, the digestion took 16 hours to complete, with ideal results observed at 24 hours. Even after extending the time to 64 hours, no non-specific digestion was detected, indicating good specificity of the enzyme under these conditions. The digestion at 37°C is shown below:
RESULTS: At 37°C, the enzyme completely digested the substrate in just 8 hours. However, the substrate also showed signs of degradation. Notably, the control group did not degrade, suggesting that EK may exhibit some non-specific digestion after prolonged incubation beyond 8 hours. This indicates a need for careful monitoring when using the enzyme at higher temperatures.
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