F278A/F279A |
the kcat values of the mutant enzyme are decreased about 1020% with the 5'-recess-end substrate and the nick substrate |
Pyrococcus horikoshii |
F278H/F279H |
kcat value decreases 83fold for the 5'-recess-end substrate and 150fold for the nick substrate compared with the wild-type values. 454fold decrease in kcat/Km for the 5'-recess-end substrate, 80fold decrease in kcat/Km for the nick substrate. The Km-value for the endo activity with the double flap substrate is increased 4fold, the kcat/Km-value is decreased 14fold, compared with the wild-type value |
Pyrococcus horikoshii |
F278H/F279H |
the Km value for the 5'-recess-end substrate is elevated 5 times compared with the wild-type value whereas for the nick substrate the Km value of F278H/F279H is 60% the wild-type value |
Pyrococcus horikoshii |
F278L/F279L |
Km-values are lower than the wild-type values |
Pyrococcus horikoshii |
F278L/F279L |
the kcat values of the mutant enzyme are decreased about 1020% with the 5'-recess-end substrate and the nick substrate |
Pyrococcus horikoshii |
F278W/F279W |
Km-values are lower than the wild-type values |
Pyrococcus horikoshii |
F278W/F279W |
the kcat values of the mutant enzyme are decreased about 1020% with the 5'-recess-end substrate and the nick substrate |
Pyrococcus horikoshii |
F278Y/F279Y |
Km-values are lower than the wild-type values |
Pyrococcus horikoshii |
F278Y/F279Y |
the kcat value of F278Y/F279Y is restored to around 70% of that of wil-type enzyme with the 5'-recess-end substrate and the nick substrate |
Pyrococcus horikoshii |
F35A |
kcat and Km of F35A and F35L show no significant decrease compared with the wild-type values |
Pyrococcus horikoshii |
F35L |
kcat and Km of F35A and F35L show no significant decrease compared with the wild-type values |
Pyrococcus horikoshii |
F35Y |
the Km values of the mutant enzyme are about 4- and 3fold higher than the values of wild-type enzyme with the nick and 5'-recess-end substrates, respectively |
Pyrococcus horikoshii |
F79A |
for the 5'-recess-end substrate, the kcat value of the mutant enzyme decreases 71fold compared with that of wild-type.For the nick substrate, the kcat value decreases 25fold compared with that of wild-type. 58fold decrease in kcat/Km for the 5'-recess-end substrate, 75fold decrease in kcat/Km for the nick substrate. The Km-value for the endo activity with the double flap substrate is increased 7fold, the kcat/Km-value is decreased 6fold, compared with the wild-type value.For the single flap and pseudo-Y substrates, the kcat of F79A decreases 31- and 37fold, respectively, compared with that of wild-type |
Pyrococcus horikoshii |
F79H |
te kcat value of the mutant for the 5'-recess-end substrate is decreased to about 50% of the wild-type value, and the kcat value of F79H for the nick substrate is seven times lower than that of wild-type enzyme. The Km value of F79H for the 5'-recess-end substrate is 13fold higher than that of wild-type, whereas the Km value of F79H for the nick substrate is about 2 times higher than that of wild-type |
Pyrococcus horikoshii |
F79L |
for the 5'-recess-end substrate, the kcat value of the mutant enzyme is restored to 20% of the wild-type value.For the nick substrate, the kcat value of the mutant enzyme is restored to 20% of that of the wild-type enzyme. The kcat-values for of the single flap and pseudo-Y substrates decrease to 17 and 7% of the wild-type values, respectively. The Km for the single flap and pseudo-Y substrates is varied moderately, but not significantly, compared with that of wild-type enzyme |
Pyrococcus horikoshii |
F79W |
the kcat values of the mutant enzyme for the 5'-recess-end substrate and the nick substrate are restored to almost the same level as the wild-type values |
Pyrococcus horikoshii |
F79Y |
the kcat values of the mutant enzyme for the 5'-recess-end substrate and the nick substrate are restored to almost the same level as the wild-type values. The kcat-value for the single flap and pseudo-Y substrates are restored to almost the same level as the wild-type substrates |
Pyrococcus horikoshii |
Y33A |
kcat decreases 333fold, compared with that of the wild-type enzyme, for exo-activity against the 5'-recess-end substrate. For the exo-activity against the nick substrate, the kcat values decreases 53fold.The kcat of Y33A for the single flap substrate decreases 30fold. The kcat of Y33A for the pseudo-Y substrate decreases 485fold |
Pyrococcus horikoshii |
Y33F |
kcat for exo-activity against the 5'-recess-end substrate is 20-30% of the wild-type value. The kcat values of Y33F for the single flap and pseudo-Y substrates are restored to 38 and 20% of the value of wild-type, respectively |
Pyrococcus horikoshii |
Y33H |
kcat for exo-activity against the 5'-recess-end substrate is 20-30% of the wild-type value |
Pyrococcus horikoshii |
Y33L |
kcat decreases 1180fold, compared with that of the wild-type enzyme, for exo-activity against the 5'-recess-end substrate. For the exo-activity against the nick substrate, the kcat values decreases 134fold. 2353fold decrease in kcat/Km for the 5'-recess-end substrate, 270fold decrease in kcat/Km for the nick substrate. The Km-value for the endo activity with the double flap substrate is increased 4fold, the kcat/Km-value is decreased 5fold, compared with the wild-type value. The kcat of Y33A for the single flap substrate decreases 433fold. The kcat of Y33A for the pseudo-Y substrate decreases 3233fold |
Pyrococcus horikoshii |
Y33W |
kcat for exo-activity against the 5'-recess-end substrate is 20-30% of the wild-type value |
Pyrococcus horikoshii |