rubrum Fed-batch culture supernatants at OD = 50. Chemical structures and molecular weights (Mw) of identified AHLs are Decitabine in vivo indicated (for a list of measured m/z values see supporting material). Single peaks were isolated by semi-preparative
HPLC and applied to A. tumefaciens NTL4 on agar plates. The inserts show the biological activity as blue colour reaction. Volume of HPLC eluate loaded onto agar containing A. tumefaciens is indicated in μL. AHL profiles at different growth modes Since R. rubrum is a very versatile life-form capable of growing under anaerobic photosynthetic conditions as well as aerobically and microaerobically in the dark, we analyzed whether the different growth modes would be reflected in the AHL profiles (for details of growth conditions see Materials and Methods). Figure 5 presents relative AHL levels in the various cultures during exponential growth. To investigate if the inhibition of PM was correlated with the AHL profile, we extracted the AHLs at two points under microaerobic growth conditions: MAE indicates extraction during PM production and MAE* indicates extraction from an older
MAE Fed-Batch culture when PM synthesis www.selleckchem.com/products/AZD6244.html was already inhibited. Figure 5 AHL accumulation profiles of R. rubrum cultivated under different growth conditions. AHL levels obtained from HPLC analysis are given in mAUsOD-1 ml-1 and are therefore qualitative estimates. AHLs were extracted from supernatants of cultures grown under phototrophic (PHO), aerobic (AE) and microaerobic (MAER) conditions. For microaerobic cultures, the supernatant was harvested at two time points. MAER* refers to a later harvesting point at which PM production has stagnated. Cultivations under aerobic and microaerobic conditions were performed in bioreactors, whereas phototrophic
cultures were grown in pyrex bottles. At top of graph, values indicate PM levels at harvest. STK38 PM value of 1.2 represents maximum PM levels and a value of 0.54 indicates a complete lack of PM formation. Strikingly, C8OH-HSL was the most abundant AHL in microaerobic cultures (Figure 5), and the sole AHL which was particularly abundant at later stages of the culture when PM production was already halted (MAE*). In phototrophic cultures with full PM expression, C8OH-HSL was the least abundant of all AHLs. In sharp contrast, C6OH-HSL was much higher in photosynthetic cultures than in microaerobic HCD cultures with repressed PM biosynthesis. C10OH-HSL was the only molecular species, elevated in PM-producing microaerobic (MAE) cultures. C8-HSL was present in all growth conditions in similar amounts except in microaerobic (MAE*) cultures where it was much lower. However, unlike the bioreactor cultivations in which the pH was stable, the pH in flask cultivations increased to ~8, which may alter stability of AHLs [23]. Accordingly, we observed differences in C6OH-HSL and C8OH-HSL accumulation between flask and bioreactor cultivations.