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A Customizable Approach for the Enzymatic Production and Purification of Diterpenoid Natural Products
JoVE 杂志
生物化学
This content is Free Access.
JoVE 杂志 生物化学
A Customizable Approach for the Enzymatic Production and Purification of Diterpenoid Natural Products

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

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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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