Search bioRxiv⌕ Search
Home/Archive/bioRxiv

bioRxiv · 10.64898/2026.07.22.740093

The Testing and Characterization of the Bacterial Response to Novel Lipid Ether Amine-Based Antibiotics

Gibson, D. J.·Svetlov, A. S.
View original source ↗Export metadata ↓

Abstract

Bacterial biofilms exhibit enhanced resistance to antibiotics compared to planktonic bacteria, though the mechanisms underlying this tolerance remain incompletely understood. Evidence suggests biofilm-associated bacteria maintain active lipid metabolism despite reduced metabolic rates, making lipid-targeting strategies potentially effective. We investigated isopropyl lipid ether amines (LEAs), computationally designed to bind phospholipase A2, as novel antibiotics targeting biofilm bacteria through lipid metabolism disruption. LEAs consist of variable-length alkyl chain analogs of natural fatty acids connected via ether linkages to cationic head groups. Using methicillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa in collagen microplate assays, minimal inhibitory concentration studies, and ex vivo porcine biofilm models, we systematically compared LEAs against their fatty acid analogs to isolate the contribution of the lipid and amine moieties. Palmitic acid analog LEA-160 and oleic acid analog LEA-181 achieved MIC and reduced microplate biofilm colony forming units (CFU) against MRSA, while short lipid chain octanoic acid analog LEA-80 reduced Pseudomonas biofilms, with no effect for natural fatty acids. This demonstrates that the cationic amine group provides essential antibacterial function beyond the alkyl chain contribution. Comprehensive lipidomics analysis using LC-IMS-MS/MS revealed that LEA treatment induces significant alterations in MRSA lipid profiles, supporting a mechanism involving disruption of bacterial membrane lipid metabolism. Structure-activity relationships confirmed that both the lipid chain and cationic moieties are necessary for LEA antibacterial efficacy, with metabolic effects distinct from their natural fatty acid analogs. These findings establish LEAs as a mechanistically distinct antibiotic class targeting bacterial lipid metabolism pathways critical for biofilm survival.

Explore related subjects

microbiologyPreprint

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Gibson, D. J., Svetlov, A. S.. 2026-07-23. The Testing and Characterization of the Bacterial Response to Novel Lipid Ether Amine-Based Antibiotics. https://doi.org/10.64898/2026.07.22.740093

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

bioRxiv2026

Matrix-controlled emergence of biofilm architecture shapes antimicrobial survival

Biofilms are structured microbial communities whose extracellular matrix is widely regarded as a basis of their protection against antimicrobial compounds. Yet how matrix production by individual bacteria gives rise to collective architecture and antimicrobial protection remains poorly understood. Here, we systematically varied expression of the master biofilm regulator csgD in Salmonella enterica and found that increasing matrix production reorganizes biofilms from dense, isotropic packings into sparse, nematically aligned communities by altering cell-cell interactions. By combining experimentally measured biofilm architectures with reaction-diffusion modeling, we show that these structural changes produce distinct patterns of antimicrobial killing, ranging from preferential killing near the liquid-biofilm interface to more uniform killing throughout the community. Consequently, increasing matrix production unexpectedly reduces antimicrobial survival by shifting the biofilm into different transport regimes, while strain-specific physiological differences further modulate antimicrobial depletion. Rather than acting as a passive barrier, EPS therefore shapes antimicrobial susceptibility by reorganizing biofilm architecture and its transport properties. EPS thus provides a physical link between molecular regulation, collective architecture and antimicrobial survival, providing a quantitative framework for understanding how cellular matrix production generates emergent biofilm function.

microbiology↗
bioRxiv2026

Mapping virulence-associated protein interaction networks reveals regulators of thermotolerance in Cryptococcus neoformans

Protein-protein interactions (PPIs) influence critical biological processes in pathogenic microorganisms, such as the human fungal pathogen, Cryptococcus neoformans. Fungal thermotolerance and stress response pathways are key virulence determinants that directly impact pathogen adaptation and survival and the infection process. To establish a comprehensive baseline of PPIs in C. neoformans and explore these interactions to infer functional roles for uncharacterized proteins, we applied size exclusion chromatography coupled with mass spectrometry to the secreted and cellular proteomes of the fungi. As a result, 216 and 1699 unique proteins were identified across 24 secretome and proteome fractions, respectively. The predicted secretome networks included expected proteins associated with vesicles and virulence, indicating a role in extracellular defense. Whereas the cryptococcal proteome highlighted interactions among proteins with defined roles in fungal virulence for protein stability and thermotolerance, including two previously uncharacterized proteins, CNAG_00287 and CNAG_05199, putatively involved in complex formation with heat-shock proteins (HSP). Based on sequence and structure homology, we propose that CNAG_00287 is a tetratricopeptide repeat-containing co-chaperone that modulates Hsp 70 activity and CNAG_05199 functions as a Hsp70. We validated the thermotolerance role of CNAG_00287 in heat-related stress, as its absence significantly impaired fungal growth in nutrient-limited media at 37 {degrees}C. Together, this work resolves virulence-associated PPIs within C. neoformans and reveals new molecular regulators of thermotolerance that underpin fungal pathogenicity.

microbiology↗
bioRxiv2026

Environmental filtering and host identity collectively shape root-associated microbiomes of Ericaceae and ectomycorrhizal plants in fumarole fields

Background Symbiosis with microbes is a key strategy that has enabled plants to colonize extreme environments. Since the benefits conferred by root-associated microbes depend on both environmental conditions and host-microbe combinations, plant adaptation to harsh environments is closely linked to the assembly of root microbial communities. Understanding how environmental and host filtering jointly shape these communities is therefore fundamental to elucidating the mechanisms underlying plant adaptation to extreme environments. Results In this study, we investigated the differentiation of root-associated prokaryotic and fungal communities and individual operational taxonomic units (OTUs) across two contrasting habitats surrounding fumaroles, solfatara-field and forest-edge habitats, and six dominant Ericaceae and ectomycorrhizal plant taxa. Prokaryotic and fungal OTUs rarely exhibited strong preferences for both habitat and host identity. Instead, many of prokaryotic and fungal OTUs specialized to one of these niches, collectively generating root microbial communities differentiated by both factors. Nonetheless, striking specializations in habitat and host niches were observed in the fungal family Hyaloscyphaceae (Helotiales). To gain insight into the evolutionary basis of microbial specialization, we examined phylogenetic signals in preference phenotypes. The resulting weak phylogenetic signals in these preference phenotypes further suggest that this fungal clade has undergone substantial ecological divergence. Conclusion Overall, our findings indicate that root-associated microbial communities in extreme environments are assembled through the accumulation of microbial taxa specialized to either habitat or host, and that strong ecological specialization in fungi can arise with little phylogenetic constraint.

microbiology↗