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Decaprenyl diphosphate synthesis in Mycobacterium tuberculosis.
Devinder Kaur1, Patrick J Brennan, Dean C Crick
1Mycobacterial Research Laboratories, Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, CO 80523-1682, USA.
Journal of Bacteriology
|November 2, 2004
Summary
Mycobacterium tuberculosis Rv2361c encodes a unique Z-prenyl diphosphate synthase. This enzyme produces decaprenyl diphosphate, a key precursor for essential cell wall components, and preferentially uses Z-configured substrates.
Area of Science:
- Biochemistry
- Molecular Biology
- Microbiology
Background:
- Z-prenyl diphosphate synthases are crucial for synthesizing polyprenyl diphosphates.
- In mycobacteria, these are precursors to decaprenyl phosphate, vital for cell wall integrity.
- Rv2361c from Mycobacterium tuberculosis was identified as a potential unique prenyl diphosphate synthase.
Purpose of the Study:
- To purify and characterize the enzyme encoded by Rv2361c.
- To determine the product of the Rv2361c enzyme.
- To elucidate the substrate specificity and optimal conditions for the enzyme's activity.
Main Methods:
- Expression of Rv2361c in Escherichia coli.
- Purification of the enzyme to near homogeneity.
- Enzyme activity assays with various allylic diphosphate substrates and isopentenyl diphosphate.
Main Results:
- The purified Rv2361c enzyme produces decaprenyl diphosphate.
- The enzyme requires divalent cations, with optimal activity at pH 7.5-8.5.
- Catalytic efficiency is highest with omega,E,Z-farnesyl diphosphate, indicating it as the likely in vivo substrate.
Conclusions:
- Rv2361c is a bacterial Z-prenyl diphosphate synthase with a preference for Z-configured allylic diphosphate primers.
- Rv1086 and Rv2361c likely act sequentially in the biosynthesis of decaprenyl phosphate in M. tuberculosis.
- This finding provides insight into the unique cell wall biosynthesis pathway of mycobacteria.