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Related Experiment Video

Updated: Jun 10, 2026

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

Published on: October 4, 2019

Diterpene production in Mycobacterium tuberculosis.

Lisa Prach¹, James Kirby, Jay D Keasling

¹Department of Molecular and Cell Biology, University of California, Berkeley, CA, USA.

The FEBS Journal

|July 31, 2010

Summary

Mycobacterium tuberculosis produces novel diterpenes, including edaxadiene and edaxadiene B, potentially by arresting phagolysosome maturation. These compounds, synthesized by unique enzymes, offer new therapeutic targets.

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Area of Science:

Microbiology
Natural Product Chemistry
Biochemistry

Background:

Diterpenes are abundant in plants but rare in bacteria.
Mycobacterium tuberculosis (Mtb) harbors unique diterpenes potentially involved in host-pathogen interactions.
Phagolysosome maturation arrest is a key mechanism in tuberculosis pathogenesis.

Purpose of the Study:

Identify and characterize novel diterpenes in Mtb.
Elucidate the biosynthetic pathway and enzymes responsible for diterpene production in Mtb.
Evaluate the potential of these diterpenes and their biosynthetic enzymes as therapeutic or diagnostic targets.

Main Methods:

Mass spectrometry (MS) for diterpene identification and analysis.
In vitro enzymatic assays using geranylgeranyl diphosphate.
Genetic manipulation of Mtb and M. smegmatis (Rv3377c and Rv3378c gene expression).

Main Results:

Three novel diterpenes identified in Mtb: edaxadiene, edaxadiene B, and tuberculosinol.
Edaxadiene and edaxadiene B are synthesized in vitro by enzymes Rv3377c and Rv3378c.
A frameshift mutation in Rv3377c abolished diterpene production in Mtb.
Expression of Rv3377c and Rv3378c in M. smegmatis produced edaxadiene and edaxadiene B, defining the in vivo pathway.
Rv3377c and Rv3378c are unique to Mtb and M. bovis.

Conclusions:

The biosynthetic pathway for edaxadiene and edaxadiene B in Mtb has been defined.
The enzymes Rv3377c and Rv3378c are responsible for the production of these novel diterpenes.
These unique enzymes represent promising targets for the development of selective therapeutics and diagnostics against Mtb and M. bovis.