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Expression and Purification of Nuclease-Free Oxygen Scavenger Protocatechuate 3,4-Dioxygenase
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Biochemistry
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JoVE Journal Biochemistry
Expression and Purification of Nuclease-Free Oxygen Scavenger Protocatechuate 3,4-Dioxygenase

Expression and Purification of Nuclease-Free Oxygen Scavenger Protocatechuate 3,4-Dioxygenase

DOI:
10.3791/59599-v

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10:14 min

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November 08, 2019

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, , ,
1Cancer Biology and Genetics, College of Medicine,The Ohio State University

Chapters

  • 00:04Title
  • 00:25Inducing PCD (protocatechuate 3,4 dioxygenase) Expression in E. coli
  • 03:39Nickel Affinity Chromatography Purification of PCD
  • 04:46Nuclease Activity Assay
  • 06:19PCA (Protocatechuic Acid) Oxidation and Nuclease Activity Assays
  • 08:01Results: Purification of PCD
  • 09:23Conclusion

Summary

Automatic Translation

Automatic Translation

Protocatechuate 3,4-dioxygenase (PCD) can enzymatically remove free diatomic oxygen from an aqueous system using its substrate protocatechuic acid (PCA). This protocol describes the expression, purification, and activity analysis of this oxygen scavenging enzyme.

Tags

Nuclease-freeOxygen ScavengerProtocatechuate 34-dioxygenasePCD ExpressionE. Coli BL21TransformationLB-ampicillin AgarProtein ExpressionIPTGAmmonium Iron SulfateCentrifugationSDS-PAGE

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