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Anaerobic Protein Purification and Kinetic Analysis via Oxygen Electrode for Studying DesB Dioxygenase Activity and Inhibition
JoVE Journal
Biochemie
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JoVE Journal Biochemie
Anaerobic Protein Purification and Kinetic Analysis via Oxygen Electrode for Studying DesB Dioxygenase Activity and Inhibition
DOI:

08:31 min

October 03, 2018

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Kapitel

  • 00:04Titel
  • 00:33Anaerobic DesB Purification
  • 02:29Anaerobic Protein Concentration, Buffer Exchange, and DesB Desalting
  • 04:10Oxygen Electrode Preparation
  • 05:41Oxygen Electrode Kinetic Assay
  • 06:48Results: Representative DesB Dioxygenase Induction, Activity, and Inhibition
  • 07:50Conclusion

Summary

Automatische Übersetzung

Here we present a protocol for anaerobic protein purification, anaerobic protein concentration, and subsequent kinetic characterization using an oxygen electrode system. The method is illustrated using the enzyme DesB, a dioxygenase enzyme which is more stable and active when purified and stored in an anaerobic environment.

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