Gunawan, C. | bioRxiv Science

bioRxiv2025

Molecular evolution of silver nanoparticle resistance in a bacterial pathogen and a unique adaptation response to ionic silver

This research explores the adaptive defense mechanisms of a nanosilver-resistant pathogen (NAgR) to protect and fight off the complex antimicrobial targeting of silver nanoparticles. The Gram-negative bacterium Acinetobacter baumannii upregulated expression of outer membrane proteins for cell surface defense, as well as membrane and capsule synthesis genes. Increased abundance of surface-attached biofilm colonies in NAgR was linked to the phenotypically indicated increase in membrane integrity, with the bacterium also forming more EPS, the biopolymer matrix that protects the residing colony. In response to the known reactive oxygen species (ROS) toxicity characteristics of the nanoparticle, NAgR upregulated its oxidative stress management system, specifically involving ROS scavenger enzymes and opportunistic metal efflux pumps. Many of these evolved defense mechanisms only manifested in the resistant bacterium, while they were absent in the wild-type strain. This study also details the unique defenses of an ionic silver-adapted A. baumannii variant, having evolved from the same wild-type parental strain as NAgR. Despite similarities in cell surface and biofilm defense trends, the slower-to-kill tolerant strain (AgT) exclusively upregulated multidrug efflux systems and respiratory chain enzymes, thought to maintain enhanced respiration activity, a known tolerant characteristic. Identification of these stable defense mechanisms can recommend strategies for molecular targeting to overcome the adaptation phenomena.

microbiology↗

bioRxiv2024

Novel sulfatase cancer therapeutics negatively impact Bacteroidota of the colonic microbiota in a non-sulfatase dependent manner

Excessive degradation of the colonic mucin layer by Bacteroides within the human gut microbiota drives inflammatory bowel disease in mice. Bacterial carbohydrate sulfatases are key enzymes in gut colonization, as they are elevated in human inflammatory bowel disease and correlate with disease severity. Selective inhibitors of carbohydrate sulfatases could function as sulfatase-selective drugs, allowing precise control of sulfatase activity while preserving these otherwise beneficial bacteria. Arylsulfamates are covalent inhibitors that target a catalytic formylglycine residue of steroid sulfatases, a residue that is also conserved in carbohydrate sulfatases. Here, we find that a library of aryl- and carbohydrate sulfamates is ineffective against Bacteroides carbohydrate sulfatases, yet can inhibit human gut microbiota species grown on sulfated glycans. Leveraging thermal proteome profiling, we identify a lipid kinase as the target responsible for these effects. This work highlights the imperative for developing specific inhibitors targeting carbohydrate sulfatases and reveals the adverse effects that arylsulfamates have on Bacteroides species of the human gut microbiota. Significance statementArylsulfamates are currently the only effective class of sulfatase inhibitors available and offer a potential strategy to treat inflammatory bowel disease driven by gut microbiota carbohydrate sulfatases. Although arylsulfamates inhibit the growth of microbiota Bacteroides species on sulfated glycans, this is not mediated through carbohydrate sulfatases but, via a conserved lipid kinase. Carbohydrate sulfatases are resistant to arylsulfamates whilst steroid sulfatases are susceptible despite a conserved active site. Finally, selected complex plant glycans confer a resistant/protective phenotype against the harmful effects of arylsulfamates. These data guide the future development of targeted carbohydrate sulfatase inhibitors and potential drug-prebiotic pairings.

biochemistry↗