# Probing the architecture, assembly, and function of amyloid-polysaccharide entanglements in bacterial biofilms

> **NIH NIH F32** · STANFORD UNIVERSITY · 2024 · $74,284

## Abstract

Project Summary
Bacteria are most commonly found in nature in multicellular communities termed biofilms. Biofilms are formed
when bacteria synthesize, secrete, and enmesh themselves with diverse biopolymers. Beneficial bacteria in the
microbiome assemble biofilms, while biofilms are unfortunately also linked to difficult-to-treat infections that
exhibit increased tolerance to antibacterials and can exhaust treatment options. However, there are no blueprints
for how bacteria build these tissue-like architectures and uncovering these details can accelerate discovery of
new anti-infectives. E. coli, in particular, are normal residents in the healthy microbiome, but emerge as
pathogens when they egress and colonize the urinary tract. E. coli, Salmonella species and other Gram-negative
organisms harness specific amyloid and polysaccharide machinery to elaborate mechanically robust
extracellular matrix architectures resembling baskets and blankets that surround cells and drive the formation of
tissue-like biofilms. Due to the complexity of biopolymer composites, there are significant challenges associated
with studying their structure and function, yet the ubiquity of these biopolymers makes them of high importance
for study. This research plan is directed to test molecular hypotheses for how bacteria employ curli and
phosphoethanolamine cellulose, a newly discovered chemically modified form of cellulose, to enmesh
themselves in extracellular matrix (ECM). The research plan will test hypotheses regarding functional roles that
we propose are ascribed to the zwitterionic phosphoethanolamine modification. Aim 1 is directed to evaluate the
temporal and spatial developments of matrix assembly beyond the bacterial cell surface using fluorescence
microscopy and creative functional biochemical assays. Aim 2 will implement a strategically designed solid-state
nuclear magnetic resonance (NMR) approach to detect molecular contacts between polysaccharides and protein
amyloids that are responsible for matrix cohesion. The functional benefit of ECM biopolymers will be determined
in Aim 3, where clinically relevant antibiotics and a novel vancomycin-conjugate will be evaluated for efficacy
against pEtN cellulose and curli containing biofilms. This work promises to formulate a molecular foundation for
future avenues of inquiry at the host-pathogen interface, involving possible immunomodulatory roles of bacterial
polysaccharides and amyloids, and possible biopolymer contributions to microbiome symbiosis and amyloid-
associated disease pathologies. The fellowship candidate will receive significant training in solid-state NMR
spectroscopy to study molecular interactions within E. coli biofilms and biochemical approaches to investigate
bacterial communities. The considerable support and mentorship structure provided through this fellowship, the
research sponsor (Prof. Lynette Cegelski) and institution (Stanford University) will facilitate the professional
development of t...

## Key facts

- **NIH application ID:** 10816991
- **Project number:** 5F32GM149117-02
- **Recipient organization:** STANFORD UNIVERSITY
- **Principal Investigator:** Schuyler A. Chambers
- **Activity code:** F32 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $74,284
- **Award type:** 5
- **Project period:** 2023-03-01 → 2025-02-28

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10816991

## Citation

> US National Institutes of Health, RePORTER application 10816991, Probing the architecture, assembly, and function of amyloid-polysaccharide entanglements in bacterial biofilms (5F32GM149117-02). Retrieved via AI Analytics 2026-06-12 from https://api.ai-analytics.org/grant/nih/10816991. Licensed CC0.

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