L homocysteine

The strain S. haloaromaticamans used in the current study was isolated from soil of a wastewater disposal site and it was able to degrade the fungicide OPP only when supplemented with CA (Perruchon et al., 2016 (link)). The bacterium was routinely cultivated in minimal salts media supplemented with nitrogen (MSMN), OPP (30–50 mg L^–1) and CA in a shaking incubator at 27°C in the dark. MSMN preparation and OPP chromatographic analysis was as described by Perruchon et al. (2016) (link), while bacterial growth was determined by measurement of the optical density at 600 nm (OD₆₀₀). Inoculation of flasks was performed by fresh bacterial cells grown at the mid-log phase, pelleted by centrifugation, washed three times with sterile ddH₂O and resuspended with MSMN to an OD₆₀₀ of 0.1.
L-methionine, L-isoleucine, L-tyrosine, L-phenylalanine, L-homoserine, O-succinyl-L-homoserine, L-cystathionine, L-homocysteine and cyanocobalamin (synonym to vitamin B12 in the manuscript), were purchased by Sigma-Aldrich (Taufkirchen, Germany) and they were used for the preparation of aqueous solutions (0.05 mM). These were filter sterilized and used for the preparation of MSMN containing amino acids, B12 and intermediates of methionine biosynthesis at the desired concentrations.

All cell culture materials were purchased from Gibco (Life Technology, Carlsbad CA, USA), and all chemicals, as well as PTH, indoxyl sulfate, ADMA, L-homocysteine and Diallyl disulfide (DADS) were from Sigma Chemical Co. (St. Louis, MO, USA), when not otherwise specified. Diallyl trisulfide (DATS) was from Cayman Chemical Co. (La Jolla, Ca, U.S.A.). PrestoBlue Cell Viability Reagent was purchased from Invitrogen (Life Technology, Carlsbad, CA, USA). pCS and pCG were synthesized at the organic chemistry department of the Ghent University, Belgium. pCS was synthesized according to Feigenbaum and Neuberg as a potassium salt [35 (link)] and pCG was synthesized and purified based on the work of Van der Eycken et al [36 (link)]. All reagents used were of analytical grade. Vacutainer tubes were purchased from Becton Dickinson Italia S.p.A.