with soil samples from agriculturally in the M sterland M sterland region. Errorindicate typical deviation (n = three). (B)(n =base (B) MS base peak chromatogramsupernatant of a soil slurry incubated soil bars indicate regular deviation MS 3). peak chromatogram on the extracted of the extracted supernatant of a with 1 mM cholate 1 mM cholate(best) about 48 h (prime) asion chromatograms withchromatograms with all the (383 Da slurry incubated with for about 48 h for together with extracted properly as extracted ion the m/z values of HOCDA m/z values for [M-H]-1, middle) and DOCDA (XX, 385 Da for [M-H]-1, bottom). Samples were measured in damaging MS mode. (C) 3D of HOCDA (383 Da for [M-H]-1 , middle) and DOCDA (XX, 385 Da for [M-H]-1 , bottom). Samples had been measured in UV chromatogram with the extracted supernatant of a soil slurry incubated with 1 mM cholate for about 48 h and structure adverse MS mode. various intermediates assigned to peaks. Intensity is shown as aa soilmap. Red indicates with 1 mM cholate ideas for (C) 3D UV chromatogram with the extracted supernatant of heat slurry incubated highest absorpfor about (D)h and structure suggestions for numerous in (B,C). Massesassigned to peaks. Intensity) is shown as a heat map. tion. 48 Candidate structures for peaks a-i located intermediates and absorption CB1 Antagonist Formulation maxima (max have been determined by HPLC-MS measurements. Structure ideas are based for peaks a-i discovered absorption spectra, and retention maxima Red indicates highest absorption. (D) Candidate structures on molecular masses, in (B,C). Masses and absorptiontime. 1,4 4,6 (maxCandidate structures by HPLC-MS measurements. Structure to the -pathway, and on molecular masses, absorption ) have been determined belonging (blue) to the -pathway, (red) ideas are primarily based (black) potentially occurring in each pathways. When structures could not be assigned CA I Inhibitor Formulation unambiguously, 1,4 feasible structures are4,6 two depicted. XV: 7,12spectra, and retention time. Candidate structures belonging (blue) towards the -pathway, (red) for the -pathway, and (black) Dihydroxy-3-oxo-pregna-4-ene-carboxylate, XVI: 7-Hydroxy-3,12-dioxo-pregna-4-ene-carboxylate, XVII: 7,12-Dihydroxypotentially occurring in each pathways. When structures could XIX: be assigned unambiguously, two achievable structures are 3-oxo-pregna-4-ene-carboxylate, XVIII: 4-3,12-Diketocholate, not DOCDA (12-Hydroxy-3-oxo-pregna-4,6-diene-carboxdepicted. XV: 7,12-Dihydroxy-3-oxo-pregna-4-ene-carboxylate, XVI: 7-Hydroxy-3,12-dioxo-pregna-4-ene-carboxylate, XVII: ylate, XX: 3,12-Dioxo-4,6-choldienoate). 7,12-Dihydroxy-3-oxo-pregna-4-ene-carboxylate, XVIII: 4 -3,12-Diketocholate, XIX: DOCDA (12-Hydroxy-3-oxo-pregna-4,64. Discussion diene-carboxylate, XX: three,12-Dioxo-4,6-choldienoate). Aerobic bacterial degradation of 7-hydroxy bile salts in soil and water can proceed via two pathway variants, namely the 1,4-variant and also the 4,6-variant [6]. The four,6-variantMicroorganisms 2021, 9,15 of4. Discussion Aerobic bacterial degradation of 7-hydroxy bile salts in soil and water can proceed via two pathway variants, namely the 1,4 -variant and the four,6 -variant [6]. The four,six variant is prevalent in the Sphingomonadaceae and differs in the 1,4 -variant, which can be found in other Proteobacteria and Actinobacteria, specifically in the degradation from the side chain [11,23], whilst the cleavage on the steroid skeleton was proposed to proceed by way of 9,10-seco cleavage in each variants. In Sphingobium sp. strain Chol11, DHSATD (XI) will be the anticipated 9,