# Chemoenzymatic preparation of tagged bactoprenyl diphosphate linked sugar probes for in vitro and cellular elucidation of complex bacterial polysaccharides

> **NIH NIH F31** · UNIVERSITY OF NORTH CAROLINA CHARLOTTE · 2020 · $34,266

## Abstract

Alternative therapeutic targets have become a major topic in research with growing concerns over the
alarming rate of antibacterial resistance. In contrast, many symbiotic bacteria that aid in human health prevent
disease and are often inadvertently targeted with broad-spectrum antimicrobials. New therapeutic targets that
selectively treat pathogenic but not symbiotic bacteria are highly valuable to balance the unique role they serve
in human health. Bacterial surface polysaccharides are a crucial component to this host-microbe interaction,
and underlie both disease and commensalism. These complex glycopolymers are highly unique in
composition, even among the same bacterial species, and are not otherwise found in mammalian systems
making them excellent therapeutic targets. The difficulty of studying these systems is the inherent complexity
associated with glycopolymer composition coupled with the lack of elegant tools to ubiquitously characterize
them. This hinders the ability to identify novel targets to inhibit these pathways, or methods to reconstruct
glycopolymers for glyco-based vaccines. Robust probes that can be used to track the formation of these
complex surface polysaccharides in vitro and in live cells are vital to exploiting them for therapeutic benefit.
 The use of tagged biomolecules has been invaluable in protein and nucleic acid biochemistry. Tagged
sugar residues are the closest equivalent, however these do not preserve the native identity of the
glycopolymer. Alternatively, formation of critical surface structures such as exopolysaccharides, teichoic acids,
and O-antigens are dependent on the common lipid carrier bactoprenyl phosphate (BP) – a polyisopreniod
carbon chain. Because BP is utilized in each step during polysaccharide assembly, it would provide a means to
track the formation of these materials. Installation of foreign tags to long carbon chain BP by chemical
synthesis is nontrivial and would entail several iterations of isoprene addition over many lengthy steps.1
Alternatively, research proposed here will focus on the chemoenzymatic preparation of a series of highly
fluorescent and click-enabled tags appended to BP. Such probes will greatly expand the range of glycopolymer
applications to include low-level detection or facile addition of application-dependent moieties, such as
biotinylation for immobilization, or tags to aid in characterization, such as signal enhancing mass spectrometry
tags. These types of tagged BP substrates are also well suited to in vivo reconstitution of glycopolymers as
they occur in nature. Incorporation of these robust tools in live cell cultures is an excellent method to probe the
dynamic assembly of critical surface polysaccharides and will facilitate the development of novel therapeutics
to reproduce or disrupt these pathways.

## Key facts

- **NIH application ID:** 10003340
- **Project number:** 5F31GM130065-03
- **Recipient organization:** UNIVERSITY OF NORTH CAROLINA CHARLOTTE
- **Principal Investigator:** Amanda Reid
- **Activity code:** F31 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $34,266
- **Award type:** 5
- **Project period:** 2018-09-01 → 2021-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10003340

## Citation

> US National Institutes of Health, RePORTER application 10003340, Chemoenzymatic preparation of tagged bactoprenyl diphosphate linked sugar probes for in vitro and cellular elucidation of complex bacterial polysaccharides (5F31GM130065-03). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10003340. Licensed CC0.

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