Biology subjects

Zarkan, A.

Publications and source records attributed to Zarkan, A..

3 recordsLinked to original sources

bioRxiv2026

STRESS HISTORY ESTABLISHES A TRANSIENT TOLERANT STATE THAT SHAPES ANTIBIOTIC SURVIVAL UPON RESUSCITATION

Antibiotic treatment failure, often driven by non-genetic mechanisms such as tolerance and persistence, remains a major global health challenge. {beta}-lactams, the most widely prescribed antibiotic class, are particularly compromised by tolerance in dormant, non-growing cells; yet, how these drugs act on cells resuscitating from dormancy remains poorly understood. Here, we investigate the resuscitation phase at an unprecedented scale using Hi-DFA (High-throughput Dynamic Fate Analyser), a single-cell microfluidic platform integrating time-lapse imaging with machine-learning-based image analysis for dynamic cell-fate tracking. We identify a distinct survival strategy: a significant fraction of resuscitating cells transiently slow their growth, facilitating survival upon {beta}-lactam exposure. This transiently tolerant phenotype is considerably less frequent in unstressed, exponentially growing cells, indicating that prior starvation history predisposes cells to this state. Using simulated in vitro pharmacokinetic treatment profiles, we show that suboptimal dosing selectively enriches for this transient tolerance state. A population dynamics model built from this single-cell antibiotic-response data suggests that these transient-tolerant cells, not typical starvation-triggered persisters, may be the primary drivers of rapid population regrowth post-treatment under clinically relevant conditions. Together, our findings define a distinct class of antibiotic survival shaped by stress history and treatment profile, offering a quantitative framework for optimising antibiotic dosing strategy.

systems biology↗

bioRxiv2025

Nanoplasmonic SERS reveals previously uncharacterised indole derivative in E. coli metabolism

Tryptophanase (TnaA) is a promiscuous enzyme which regulates the production of amino acid-derived metabolites, essential for bacterial communication. TnaA is known to convert tryptophan into indole, an important signalling molecule influencing a range of behaviours - from bacterial quorum sensing to urinary tract infections (UTI) - but other conversion routes remain obscure. Here, we show that nanoplasmonic surface-enhanced Raman spectroscopy (SERS) combined with targeted genetic subtraction enables the label-free, cellular characterisation and structural identification of TnaA-derived metabolites directly from Escherichia coli culture solutions, without isolation or purification. We systematically profile the SERS spectra of wild-type and tnaA gene knockout strains (E. coli BW25113 and the uropathogenic 536) supplemented with each of the twenty amino acids, uncovering a previously unreported TnaA-dependent metabolic signature, I*, whose Raman fingerprint does not match free indole or any common indole derivative characterised by mass spectrometry. Using isotopomer-labelled substrates together with structural reference compounds, we perform label-free structural deduction of I* directly from SERS spectra. We find I*s structure is consistent with an intact indole ring carrying a C3 substituent, likely produced indirectly from intracellular tryptophan. These findings introduce a novel indole metabolite, raise new hypotheses regarding the biological role of TnaA-mediated byproducts in bacterial signalling and virulence, and demonstrate that nanoplasmonic SERS provides a powerful framework for probing enzyme activity and bioactive metabolite production directly from living bacterial cultures with nanomolar sensitivity and structural specificity.

microbiology↗

bioRxiv2018

Indole Pulse Signalling Regulates the Cytoplasmic pH of E. coli in a Memory-Like Manner

Bacterial cells are critically dependent upon pH regulation. Most proteins function over a limited pH range and the pH gradient across the bacterial cell membrane is central to energy production and transduction1. Here we demonstrate that indole plays a critical role in the regulation of the cytoplasmic pH of E. coli. Indole is an aromatic molecule with diverse signalling roles that in bacteria is produced from tryptophan by the enzyme tryptophanase (TnaA)2. Two modes of indole signalling have been described: persistent and pulse signalling. The latter is illustrated by the brief but intense elevation of intracellular indole during stationary phase entry3,4. We show that E. coli cells growing under conditions where no indole is produced maintain their cytoplasmic pH at 7.8 {+/-} 0.2. In contrast, under conditions permitting indole production, pH is maintained at 7.2 {+/-} 0.2. Experiments where indole was added experimentally to non-producing cultures showed that pH regulation results from pulse, rather than persistent, indole signalling. Furthermore, the application of an artificial pulse of either of two non-biological proton ionophores (DNP or CCCP) caused a similar effect, suggesting that the relevant property of indole in this context is its ability to conduct protons across the cytoplasmic membrane5. Additionally, we show that the effect of the indole pulse that occurs normally during stationary phase entry in rich medium remains as a \"memory\" to maintain the correct cytoplasmic pH until entry into the next stationary phase. The indole-mediated reduction in cytoplasmic pH may explain why indole provides E. coli with a degree of protection against stresses, including some bactericidal antibiotics.

microbiology↗