Hydrolysis of Antimicrobial Peptides by Extracellular Peptidases in Wastewater.

Several antimicrobial peptides (AMPs) are emerging as promising novel antibiotics. When released into wastewater streams after use, AMPs might be hydrolyzed and inactivated by wastewater peptidases─resulting in a reduced release of active antimicrobials into wastewater-receiving environments. A key step towards a better understanding of the fate of AMPs in wastewater systems is to investigate the activity and specificity of wastewater peptidases. Here, we quantified peptidase activity in extracellular extracts from different stages throughout the wastewater treatment process. For all four tested municipal wastewater treatment plants, we detected highest activity in raw wastewater. Complementarily, we assessed the potential of enzymes in raw wastewater extracts to biotransform 10 selected AMPs. We found large variations in the susceptibility of AMPs to enzymatic transformation, indicating substantial substrate specificity of extracted enzymes. To obtain insights into peptidase specificities, we searched for hydrolysis products of rapidly biotransformed AMPs and quantified selected products using synthetic standards. We found that hydrolysis occurred at specific sites and that these sites were remarkably conserved across the four tested wastewaters. Together, these findings provide insights into the fate of AMPs in wastewater systems and can inform the selection and design of peptide-based antibiotics that are hydrolyzable by wastewater peptidases.

Photochemical Production of Carbon Monoxide from Dissolved Organic Matter: Role of Lignin Methoxyarene Functional Groups.

Carbon monoxide (CO) is the second most abundant identified product of dissolved organic matter (DOM) photodegradation after CO2, but its formation mechanism remains unknown. Previous work showed that aqueous photodegradation of methoxy-substituted aromatics (ArOCH3) produces CO considerably more efficiently than aromatic carbonyls. Following on this precedent, we propose that the methoxy aromatic groups of lignin act as the C source for the photochemical formation of CO from terrestrial DOM via a two-step pathway: formal hydrolytic demethylation to methanol and methanol oxidation to CO. To test the reasonableness of this mechanism, we investigated the photochemistry of eight lignin model compounds. We first observed that initial CO production rates are positively correlated with initial substrate degradation rates only for models containing at least one ArOCH3 group, regardless of other structural features. We then confirmed that all ArOCH3-containing substrates undergo formal hydrolytic demethylation by detecting methanol and the corresponding phenolic transformation products. Finally, we showed that hydroxyl radicals, likely oxidants to initiate methanol oxidation to CO, form during irradiation of all models. This work proposes an explicit mechanism linking ubiquitous, abundant, and easily quantifiable DOM functionalities to CO photoproduction. Our results further hint that methanol may be an abundant (yet overlooked) DOM photoproduct and a likely precursor of formaldehyde, formic acid, and CO2 and that lignin photodegradation may represent a source of hydroxyl radicals.

Charge Effects on the Photodegradation of Single Optically Trapped Oleic Acid Aerosol Droplets.

It has recently been reported that reactions can occur faster in microdroplets than in extended condensed matter. The electric charge of droplets has also been suggested as a possible cause of this phenomenon. Here, we investigate the influence of electric charges on the photodegradation of single, optically trapped oleic acid aerosol droplets in the absence of other reactive species. The temporal evolution of the chemical composition and the size of droplets with charge states ranging from 0 to 104 elementary charges were retrieved from Raman spectra and elastic light scattering, respectively. No influence of the droplet charge was observed, either on the chemical composition or on the kinetics. Based on a kinetic multilayer model, we propose a reaction mechanism with the photoexcitation of oleic acid into an excited state, subsequent decay into intermediates and further photoexcitation of intermediates and their decay into nonvolatile and volatile products.