The Fe Chelator Proferrorosamine A Is Essential for the Siderophore-Mediated Uptake of Iron by Pseudomonas roseus fluorescens.

Pseudomonas roseus fluorescens produces, besides the Fe chelator proferrorosamine A, Fe -chelating compounds, called siderophores. The production of proferrorosamine A and siderophores by P. roseus fluorescens appears to be controlled in a similar way by the concentration of available iron and by the concentration of dissolved oxygen. The higher the concentration of iron available for the microorganism, the lower the production of both chelating compounds. However, the production of siderophores was much more sensitive to iron availability than was proferrorosamine A production. Proferrorosamine A and siderophores were only produced in minimal medium C if the concentration of dissolved oxygen ranged from 4.5 to 2.0 ppm. At higher or lower concentrations, none of the iron-chelating compounds were produced. Furthermore, it has been shown that proferrorosamine-negative Tn5 mutants of P. roseus fluorescens were able to form siderophores only under iron-limiting conditions when proferrorosamine A was added to the medium. Our data suggest that proferrorosamine A production is essential for siderophore synthesis by P. roseus fluorescens; the production of siderophores occurred only when proferrorosamine A was present.

Iron complexation as a tool to direct mixed clostridia-lactobacilli fermentations.

Iron complexation was investigated as a possible tool to give lactobacilli a competitive advantage over clostridia. The iron complexing substance tested, i.e. 2,2'-dipyridyl, was not toxic itself for clostridia, but its addition to a mixed culture of lactobacilli and clostridia resulted in a strong ecological advantage of the lactobacilli.

Manganese as a controlling factor in mixed cultures of Lactobacillus plantarum and Enterobacteriaceae.

Addition of manganese, at levels of 50 ppm, to a liquid growth medium simulating adverse silage conditions had no effect on the growth or on the fermentation pattern of Enterobacter cloacae and Proteus vulgaris. Yet, the manganese strongly enhanced the growth of Lactobacillus plantarum. Co-cultures of L. plantarum and E. cloacae or P. vulgaris were, by addition of manganese ions, significantly altered in the favour of the former. This finding can be of use in mixed cultures where Enterobacteriaceae act as spoiler microorganisms.

Physicochemical characterization of the microbial Fe2+ chelator proferrorosamine from Pseudomonas roseus fluorescens.

The microbial chelating compound proferrorosamine A, produced by Pseudomonas roseus fluorescens, formed a complex with Fe2+ of which the apparent stability constant was found to be 10(23). The following order of increasing stability constants of metal complexes with proferrorosamine was established as: Ba2+, Ca2+, Mg2+, Mn2+ less than Hg2+ less than Zn2+ less than Pb2+ less than Co2+ less than Cu2+ congruent to Fe2+ less than Ni2+. Only Ni(2+)-proferrorosamine had a stability constant which was established as: Ba2+, Ca2+, Mg2+, Mn2+ less than Hg2+ less than Zn2+ less than Pb2+ less than Co2+ less than Cu2+ congruent to Fe2+ less than Ni2+. Only Ni(2+)-proferrorosamine had a stability constant which was ca 32 times higher than Fe(2+)-proferrorosamine. Because of the production of proferrorosamine the growth of Ps. roseus fluorescens was not inhibited in iron limiting media by the addition of 0.15 mmol/l of the weaker chemical Fe2+ chelator 2,2'-dipyridyl. This contrasted with the proferrorosamine-negative mutant K2 and Ps. stutzeri, which only produces Fe(3+)-chelating siderophores. Furthermore, it was found that proferrorosamine was able to dissolve Fe2+ from stainless steel. These results show that proferrorosamine is a strong and selective Fe2+ chelator which could be used as an alternative for the toxic 2,2'-dipyridyl to control lactic acid fermentations.

Development of a 5-step multi-chamber reactor as a simulation of the human intestinal microbial ecosystem.

A five-stage reactor was developed to simulate the gastro-intestinal microbial ecosystem of humans. The small intestine was simulated by a two-step "fill and draw" system, the large intestine by a three-step reactor. A representative supply medium was developed to support a microbial community resembling that of the human gastro-intestinal tract. The entire system was validated by monitoring fermentation fluxes and products, i.e. indicator bacterial groups, volatile fatty acids, enzymatic activities and headspace gases. The simulator was operated with varying concentrations and combinations of arabinogalactan, xylan, pectin, dextrins and starch. The resulting patterns of microbial diversity and activity were analysed and compared with data for in-vivo gastro-intestinal microbial communities as described in the literature and found to be representative.

Significance of bile salt hydrolytic activities of lactobacilli.

Bile salt hydrolase (BSH) activity was shown to be constitutive and substrate-specific: the BSH isogenic Lactobacillus plantarum wild type (LP80 WT) and BSH overproducing LP80 (pCBH1) strains preferentially hydrolysed glycodeoxycholic acid (GDCA), whereas the hamster Lact. animalis isolates H362 and H364 showed a higher affinity for taurodeoxycholic acid (TDCA). In viability studies in the presence of nutrients, it was demonstrated that GDCA exerted a higher toxicity than TDCA in a pH-dependent manner. This toxicity was inversely proportionate to the BSH activity level of the strains tested, indicating that BSH activity contributed towards bile salt resistance when appropriate nutrients were available. The high toxicity of GDCA relative to TDCA was suggested to be caused by their weak and strong acid properties respectively. It was therefore hypothesized that the protonated form of bile salts exhibited toxicity as it imported protons in the cell. This puts an energy-burden on BSH- lactobacilli which undergo intracellular acidification. BSH+ cells primarily protect themselves through the formation of the weaker DCA compound, which can help negate the pH-drop by recapturing and exporting the co-transported proton. However, since DCA is more toxic than its conjugated counterparts, an additional energy-dependent detoxification of DCA is suggested.

Role of autotrophic nitrifiers in biological manganese removal from groundwater containing manganese and ammonium.

Upon start-up of a rapid sand filter fed with groundwater containing Mn(2+) and NH(4+), the first to be removed was NH(4+), which was oxidized to NO2 (-). After both NH(4+) and NO2 (-). were completely oxidized to NO3 (-), the removal of Mn(2+) commenced. Batch experiments showed that the addition of Nitrosomonas europaea and Nitrobacter winogradskyi stimulated the Mn(2+) removal by sandfilter microbial consortia. NO2 (-). was found to have a marked inhibitory effect on the removal of Mn(2+) and could reduce the removal rate by half. In this respect, NO2 (-)-mediated chemical reduction of manganese oxide was demonstrated at slightly acidic pH values. In pure cultures of Nitrosomonas europaea and Nitrobacter winogradskyi, no Mn(2+) oxidation occurred, but reduction of MnO2 to Mn(2+) was found when NO2 (-). accumulated. These results indicate that the development of NO2/(-). oxidizers is critical in the removal of Mn(2+) in rapid sand filters. By oxidizing NO2 (-). NO2 (-). oxidizers eliminate the negative effect of NO2 (-). on the biological oxidation of Mn(2+).