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Updated: Jul 6, 2026
13:19
Chemical Triphosphorylation of Oligonucleotides
Published on: June 2, 2022
Chemoenzymatic synthesis of polyprenyl phosphates
Meredith D Hartley1, Angelyn Larkin, Barbara Imperiali
1Department of Chemistry, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139, USA.
Bioorganic & Medicinal Chemistry
|April 1, 2008
Summary
Streptococcus mutans diacylglycerol kinase (DGK) efficiently phosphorylates polyprenols for synthesizing essential polyprenyl phosphates. This enzyme aids in creating key intermediates for N-linked glycosylation and cell wall biosynthesis studies.
Area of Science:
- Biochemistry
- Enzymology
- Microbiology
Background:
- Polyprenyl phosphates are crucial for N-linked protein glycosylation and bacterial cell wall synthesis.
- Undecaprenol kinase activity is vital for producing these essential phosphorylated intermediates.
Purpose of the Study:
- To evaluate undecaprenol kinases for chemoenzymatic synthesis of polyprenyl phosphates.
- To assess the utility of Streptococcus mutans diacylglycerol kinase (DGK) for polyprenol phosphorylation.
Main Methods:
- Expression of target enzymes in crude cell envelope fractions.
- Quantification of enzymes using luminescent lanthanide-binding tags (LBTs).
- Chemoenzymatic synthesis using ATP as the phosphoryl donor.
Main Results:
- Streptococcus mutans DGK demonstrated high utility for polyprenol phosphorylation.
- Coupling S. mutans DGK with Campylobacter jejuni glycosyltransferases enabled efficient disaccharide synthesis.
- The synthesized undecaprenyl pyrophosphate-linked disaccharide is suitable for PglB studies.
Conclusions:
- S. mutans DGK is an effective biocatalyst for synthesizing polyprenyl phosphates.
- This chemoenzymatic approach facilitates the production of essential substrates for glycosylation research.
- The study provides a valuable method for investigating N-linked glycosylation pathways.
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