This gum was triturated with methanol upon which it partly

solidified. Decanting off the methanol and repeating the procedure with fresh methanol led finally to a complete solidification. The 1H-NMR spectrum, analogous to that of Roy & Hewlins (1997), showed an enrichment of SQ as a mixture of its anomers over p-toluenesulfonic acid (≤ 10%) and no other organic impurities. Data from MALDI-TOF-MS in the negative ion mode gave m/z = 443 = [M−1]−1, which is consistent with SQ (M = 444). The syntheses of DHPS and racemic sulfolactate were described elsewhere (Roy et al., 2003; Mayer et al., 2010). Other chemicals were available commercially from Sigma-Aldrich, Fluka, Merck or Biomol. Burkholderia phymatum STM815 (DSM 17167) (e.g. Elliott et al., 2007), Burkholderia xenovorans LB400 (e.g. Chain et al., 2006), Cupriavidus necator H16 (DSM 428) (e.g. Pohlmann et al., 2006), Cupriavidus selleck chemicals llc pinatubonensis JMP134 (DSM 4058) (Sato et al., 2006), K. oxytoca TauN1 (DSM 16963) (Styp von Rekowski see more et al., 2005), Paracoccus pantotrophus NKNCYSA (DSM 12449) (e.g. Rein et al., 2005), Sinorhizobium meliloti Rm1021 (e.g. Finan et al., 2001), Rhodopseudomonas palustris CGA009 (e.g. Larimer et al., 2004), Rhodobacter sphaeroides

2.4.1 (e.g. Mackenzie et al., 2001), P. putida F1 (e.g. Zylstra & Gibson, 1989), and P. putida KT2440 (e.g. Nelson et al., 2002) were grown aerobically at 30 °C in a phosphate-buffered mineral salts medium, pH 7.2 (Thurnheer et al., 1986). Roseobacter

litoralis Och 149 (DSM 6996) (e.g. Kalhoefer et al., 2011) and Roseovarius sp. those strain 217 (Schäfer et al., 2005) were cultured in a Tris-buffered artificial seawater medium (Krejčík et al., 2008). Strain Och 149 was grown at 25 °C and strain 217 required the addition of vitamins (Pfennig, 1978). Roseovarius nubinhibens ISM (González et al., 2003) was grown in modified Silicibacter basal medium (Denger et al., 2006) and needed a supplement of 0.05% yeast extract (Denger et al., 2009). The sole carbon source was 5 mM sulfoquinovose or as a control 20 mM acetate or taurine or 10 mM succinate or 5 mM 4-toluenesulfonate or 5 mM glucose. Cultures on the 3-mL scale in 30-mL screw-cap tubes were incubated in a roller. For growth experiments, 12-mL cultures were grown in a beaker on a shaker, and 0.8 mL samples were taken at intervals to measure the optical density at 580 nm and to analyze concentrations of substrate and product. Enrichment cultures were set up in a 3-mL scale in the freshwater mineral salts medium with 5 mM SQ as sole added carbon source. If turbidity developed and bacteria could be seen under the microscope, subcultures in fresh selective medium were inoculated. After four or five transfers, cultures were streaked on LB-agar plates and colonies were picked into fresh selective medium. After three rounds of plating and picking from homogeneous plates, cultures were considered pure.