2025-03-062025-03-062024-09-30Masotti, F., Krink, N., Lencina, N., Gottig, N., Ottado, J., & Nikel, P. I. (2024). Disentangling the Regulatory Response of Agrobacterium tumefaciens CHLDO to Glyphosate for Engineering Whole-Cell Phosphonate Biosensors. ACS synthetic biology, 13(10), 3430–3445. https://doi.org/10.1021/acssynbio.4c004972161-5063https://hdl.handle.net/2133/29008Phosphonates (PHTs), organic compounds with a stable C−P bond, are widely distributed in nature. Glyphosate (GP), a synthetic PHT, is extensively used in agriculture and has been linked to various human health issues and environmental damage. Given the prevalence of GP, developing cost-effective, onsite methods for GP detection is key for assessing pollution and reducing exposure risks. We adopted Agrobacterium tumefaciens CHLDO, a natural GP degrader, as a host and the source of genetic parts for constructing PHT biosensors. In this bacterial species, the phn gene cluster, encoding the C−P lyase pathway, is regulated by the PhnF transcriptional repressor. We selected the phnG promoter, which displays a dose-dependent response to GP, to build a set of whole-cell biosensors. Through stepwise genetic optimization of the transcriptional cascade, we created a whole-cell biosensor capable of detecting GP in the 0.25−50 μM range in various samples, including soil and water.3430- 3445enopenAccessGlyphosateWhole-cell biosensorSynthetic biologyAgrobacteriumMetabolic engineeringBiodegradationarticuloMasotti, FiorellaKrink, NicolasLencina, NicolasGottig, NataliaOttado, JorgelinaNikel, Pablo I.Attribution-NonCommercial-NoDerivatives 4.0 International