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A Customizable Approach for the Enzymatic Production and Purification of Diterpenoid Natural Products
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
A Customizable Approach for the Enzymatic Production and Purification of Diterpenoid Natural Products
DOI:

07:59 min

October 04, 2019

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Chapters

  • 00:04Title
  • 00:37Diterpenoid Metabolite Production and Transformation in E. coli
  • 02:14Inoculation Culture Preparation
  • 02:59Co-Expression Cultures Preparation and Induction
  • 04:34Metabolite Separation and Purification
  • 05:55Results: Representative Diterpenoid Enzymatic Production and Purification Analyses
  • 07:02Conclusion

Summary

Automatic Translation

Here we present easy to use protocols for producing and purifying diterpenoid metabolites through the combinatorial expression of biosynthetic enzymes in Escherichia coli or Nicotiana benthamiana, followed by chromatographic product purification. The resulting metabolites are suitable for various studies including molecular structure characterization, enzyme functional studies, and bioactivity assays.

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