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Knoll, W.

Publications and source records attributed to Knoll, W..

2 recordsLinked to original sources

New potential antimicrobial peptides with amazing symmetrical structure in fungi and insects

We have discovered a new family of genes encoding potential antimicrobial peptides with unique compact and elegant structure in the genomes of several Fungi and some arthropod species. Their expression products are constituted of about 85 amino acids, including a signal peptide, and are folded into two -helical segments connected by a short unstructured coil. Three conserved disulphide bridges between cysteines located in symmetrically mirrored positions connect the two helical domains. The two ends of the chain are thus brought together and the elongated and compact shape suggests that of a nail, potentially able to penetrate the cell membrane. A high abundance of hydrophobic residues supports such hypothesis. These peptides, that we name Hairpin Loop Peptides (HLPs), have been found in the genomes of many Fungi species but only in selected clades. Orthologues have also been discovered in the genomes of some insects, notably Hemiptera, a few other arthropods and other organisms, but are absent in plants. They appear to have originated in Fungi and then migrated to insects through horizontal gene transfer. The antimicrobial activity of HLPs is predicted by several software programmes, although this still needs to be confirmed by experimental evidence. The occurrence of HLPs in several edible mushrooms supports potential uses of these peptides in food preservation and possibly also in medical applications. Their simple and nearly rigid structure can be easily modified to improve specificity, stability and solubility, thus making these molecular weapons suitable for a variety of different applications. Significance statementThe increasing emergence of bacteria resistant to current antibiotics has stimulated a rapid search for alternative treatments. In recent years, antimicrobial peptides have attracted considerable interest for their potential applications in medicine and as food preservatives. We have identified a new class of peptides primarily expressed in fungi, including edible mushrooms, and also detected in some insects and other arthropods, likely as a result of horizontal gene-transfer events. These peptides are particularly noteworthy because of their compact, elongated, and highly symmetrical structures, which give them a nail-like shape capable of penetrating cellular membranes. Such structural features suggest potential antimicrobial activity, a prediction supported by computational analyses. Their widespread presence in edible mushrooms further indicates the potential safety of these peptides for human use.

microbiology↗

gFET-based aptasensors technology allows sensitive and specific quantification of the ESKAPE pathogens

The rapid rise of antimicrobial resistance among the ESKAPE pathogens, Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and Enterobacter spp., requires diagnostic technologies capable of fast, simplified and sensitive microbial detection. Conventional culture-based diagnostics remain too time-consuming to guide early therapeutic interventions. Here, we present a graphene field-effect transistor (gFET) aptasensor platform enabling rapid, label-free and highly sensitive quantification of all six ESKAPE pathogens. Each gFET device was functionalized with a specific DNA aptamer selected from literature sources and immobilized via a mixed pyrene-based linker strategy on reduced graphene oxide (rGO). Exposure of the functionalized sensors to logarithmically diluted bacterial suspensions (10-105 CFU mL-) produced characteristic and concentration-dependent shifts in source-drain current ({Delta}IDS). For all pathogen-specific aptamers, {Delta}IDS correlated linearly with bacterial load (R2 = 0.90-1.00), while non-target bacteria generated only low-level, unspecific fluctuations. Limits of detection ranged from 10 to 1000 bacterial cells depending on the aptamer. Together, these results demonstrate that aptamer-functionalized rGO-FETs provide a robust, scalable and highly specific electronic biosensing architecture capable of distinguishing clinically relevant multidrug-resistant pathogens with excellent analytical performance. Author summaryAntimicrobial-resistant bacteria pose a growing threat to global health, especially the so-called ESKAPE pathogens, which frequently cause hospital-acquired infections and are increasingly difficult to treat. Current diagnostic methods can take several days, delaying the start of effective therapy. In our work, we developed a fast and highly sensitive biosensor that uses electrically conductive GO and short DNA molecules, called aptamers, to recognize specific bacteria. When a pathogen binds to its matching aptamer on the sensor surface, the electrical signal of the graphene changes in a measurable way. We tested ESKAPE species and showed that all of the investigated aptamers detect only their intended bacterial targets, even at very low concentrations. Importantly, the sensors respond within minutes and do not require any labelling or complex sample preparation. Our technology demonstrates how graphene-based aptasensors can support rapid and accurate detection of dangerous bacterial pathogens and could ultimately help clinicians make faster decisions in treating infections.

microbiology↗