TY - GEN A1 - Schwedt, Inge A1 - Schöne, Kerstin A1 - Eckert, Maike A1 - Pizzinato, Manon A1 - Winkler, Laura A1 - Knotkova, Barbora A1 - Richts, Björn A1 - Hau, Jann‐Louis A1 - Steuber, Julia A1 - Mireles, Raul A1 - Noda‐Garcia, Lianet A1 - Fritz, Günter A1 - Mittelstädt, Carolin A1 - Hertel, Robert A1 - Commichau, Fabian M. T1 - The low mutational flexibility of the EPSP synthase in Bacillus subtilis is due to a higher demand for shikimate pathway intermediates T2 - Environmental Microbiology N2 - Glyphosate (GS) inhibits the 5-enolpyruvyl-shikimate-3-phosphate (EPSP) synthase that is required for aromatic amino acid, folate and quinone biosynthesis in Bacillus subtilis and Escherichia coli. The inhibition of the EPSP synthase by GS depletes the cell of these metabolites, resulting in cell death. Here, we show that like the laboratory B. subtilis strains also environmental and undomesticated isolates adapt to GS by reducing herbicide uptake. Although B. subtilis possesses a GS-insensitive EPSP synthase, the enzyme is strongly inhibited by GS in the native environment. Moreover, the B. subtilis EPSP synthase mutant was only viable in rich medium containing menaquinone, indicating that the bacteria require a catalytically efficient EPSP synthase under nutrient-poor conditions. The dependency of B. subtilis on the EPSP synthase probably limits its evolvability. In contrast, E. coli rapidly acquires GS resistance by target modification. However, the evolution of a GS-resistant EPSP synthase under non-selective growth conditions indicates that GS resistance causes fitness costs. Therefore, in both model organisms, the proper function of the EPSP synthase is critical for the cellular viability. This study also revealed that the uptake systems for folate precursors, phenylalanine and tyrosine need to be identified and characterized in B. subtilis. Y1 - 2023 U6 - https://doi.org/10.1111/1462-2920.16518 SN - 1462-2912 SN - 1462-2920 VL - 25 IS - 12 SP - 3604 EP - 3622 ER - TY - GEN A1 - Riedel, Ramona A1 - Commichau, Fabian M. A1 - Benndorf, Dirk A1 - Hertel, Robert A1 - Holzer, Katharina A1 - Mardoukhi, Mohammad Saba Yousef A1 - Noack, Laura A1 - Martienssen, Marion T1 - Biodegradation of selected aminophosphonates by the bacterial isolate Ochrobactrum sp. BTU1 T2 - Microbial Research N2 - Aminophosphonates, like glyphosate (GS) or metal chelators such as ethylenediaminetetra(methylenephosphonic acid) (EDTMP), are released on a large scale worldwide. Here, we have characterized a bacterial strain capable of degrading synthetic aminophosphonates. The strain was isolated from LC/MS standard solution. Genome sequencing indicated that the strain belongs to the genus Ochrobactrum. Whole-genome classification using pyANI software to compute a pairwise ANI and other metrics between Brucella assemblies and Ochrobactrum contigs revealed that the bacterial strain is designated as Ochrobactrum sp. BTU1. Degradation batch tests with Ochrobactrum sp. BTU1 and the selected aminophosphonates GS, EDTMP, aminomethylphosphonic acid (AMPA), iminodi(methylene-phosphonic) (IDMP) and ethylaminobis(methylenephosphonic) acid (EABMP) showed that the strain can use all phosphonates as sole phosphorus source during phosphorus starvation. The highest growth rate was achieved with AMPA, while EDTMP and GS were least supportive for growth. Proteome analysis revealed that GS degradation is promoted by C-P lyase via the sarcosine pathway, i.e., initial cleavage at the C-P bond. We also identified C-P lyase to be responsible for degradation of EDTMP, EABMP, IDMP and AMPA. However, the identification of the metabolite ethylenediaminetri(methylenephosphonic acid) via LC/MS analysis in the test medium during EDTMP degradation indicates a different initial cleavage step as compared to GS. For EDTMP, it is evident that the initial cleavage occurs at the C-N bond. The detection of different key enzymes at regulated levels, form the bacterial proteoms during EDTMP exposure, further supports this finding. Y1 - 2024 U6 - https://doi.org/10.1016/j.micres.2024.127600 SN - 0944-5013 VL - 280 SP - 1 EP - 12 ER -