Blanchet, J. | Search bioRxiv

bioRxiv2026

Hydration and hydrolysis define antibiotic resistance conferred by macrolide esterases

Macrolides are an antibiotic class widely used in both human and veterinary medicine, and function by interfering with protein synthesis. Regrettably, numerous strategies for evading the antibiotic properties of macrolides have been found in bacteria, including enzyme-mediated inactivation. These mechanisms are now widely disseminated among pathogenic, animal-associated and environmental bacteria making them a One Health issue. Macrolide esterases, which hydrolyze the macrolactones ester bond, confer one such resistance mechanism. Two types of macrolide esterases have thus far been identified, the well-studied erythromycin esterases and the recently discovered Est-type enzymes that belong to the /{beta}-hydrolase superfamily. We present detailed structure-function studies for four diverse Est type esterases: which only share 44-66% sequence identity (EstTSf, EstTSt, EstTBc, and EstXEc). In addition to resistance profiling and substrate specificity studies, we present structures for all four enzymes, including structures for EstTBc and EstXEc in complex with tylosin and tylvalosin macrolides, post hydrolysis. Complementing the data with mutational and kinetic studies allowed for a detailed analysis of the structural basis for macrolide-enzyme interactions. Combined the data suggest that promiscuous binding and imprecise positioning, mediated by a water-cage, dictate substrate specificity for Est-type macrolide resistance enzymes. These insights may prove beneficial for next-generation antibiotic development.

biochemistry↗

bioRxiv2026

Broadleaved hedgerows as complementary habitats for small mammals in pine plantation landscapes

O_LIThe global decline of natural forests is accompanied by a rapid expansion of commercial tree plantations, which are expected to further increase to meet growing demand for wood products. However, planted forests generally support lower biodiversity than natural forests, particularly when monospecific and intensively managed. In this context, broadleaved hedgerows have been proposed as a nature-based solution to enhance biodiversity within conifer-dominated plantation landscapes. Such features may be especially beneficial for small mammals, including rodents and shrews, which are key contributors to forest ecosystem functioning. However, their effects on small mammal communities remain largely unquantified. C_LIO_LIHere, we assessed variation in small mammal communities among habitat types within a native pine plantation-dominated landscape in southwestern France. Using a multi-year, multi-season survey, we compared species richness and abundance among plantation edges, broadleaved hedgerows embedded within plantations and natural broadleaved forests. We further tested whether environmental descriptors of hedgerow sites influenced dominant species and whether seasonal and interannual demographic dynamics modified habitat-related patterns. C_LIO_LIPine plantation edges and broadleaved hedgerows supported lower small mammal species richness than natural broadleaved forests and were dominated by two habitat generalists, Apodemus sylvaticus and Crocidura russula. This pattern was driven by the near absence of the forest specialist Clethrionomys glareolus. Hedgerows did not increase species richness relative to plantations, but provided favourable habitat for A. sylvaticus, which was scarce in pine plantation, while supporting fewer C. russula. Variation in hedgerow structure and composition further influenced A. sylvaticus abundance, while seasonal and interannual rodent population dynamics modulated habitat-related differences. C_LIO_LIOur results indicate that intensively managed pine plantations act as environmental filters, excluding forest-associated small mammals. While broadleaved hedgerows benefited one species, their capacity to restore forest-specialist communities was limited without broader landscape-scale interventions. These findings highlight both the ecological benefits and constraints of edge-based habitat interventions and provide guidance for designing and evaluating biodiversity-oriented management in plantation landscapes. C_LI

ecology↗