carotenoids are associated with several important biological functions as antenna pigments in photosynthesis or protectives against oxidative stress. Occasionally they were also discussed as part of the cold adaptation mechanism of bacteria. For two Staphylococcus xylosus strains we demonstrated an increased content of staphyloxanthin and other carotenoids after growth at 10 °C but no detectable carotenoids after grow at 30 °C. By in vivo measurements of generalized polarization and anisotropy with two different probes Laurdan and TMA-DPH we detected a strong increase in membrane order with a simultaneous increase in membrane fluidity at low temperatures accompanied by a broadening of the phase transition. Increased carotenoid concentration was also correlated with an increased resistance of the cells against freeze-thaw stress. In addition, the fatty acid profile showed a moderate adaptation to low temperature by increasing the portion of anteiso-branched fatty acids. The suppression of carotenoid synthesis abolished the effects observed and thus confirmed the causative function of the carotenoids in the modulation of membrane parameters. A differential transcriptome analysis demonstrated the upregulation of genes involved in carotenoid syntheses under low temperature growth conditions. The presented data suggests that upregulated synthesis of carotenoids is a constitutive component in the cold adaptation strategy of Staphylococcus xylosus and combined with modifications of the fatty acid profile constitute the adaptation to grow under low temperature conditions. Carotenoids represent a large group and comprise at least 800 described compounds 1,2 which are synthesized from isoprene units by phototrophic and chemoorganotrophic prokaryotes, plants, fungi, and algae 3. Most carotenoids consist of eight isoprene units resulting in a C40 backbone and usually display β-cyclisation 1,4. Only a few chemoorganotrophic bacteria are capable of producing C30, C45 or C50 carotenoids 4,5. Such a rare C30 carotenoid is produced by staphylococci 6,7. As lipophilic compounds, carotenoids are located in the cell membrane, but their orientation inside the membrane may vary depending on their chemical structure and on the thickness of the membrane 8,9. They perform important biological functions such as light harvesting as antenna pigments 10,11 , provide protection against oxidative stress 5,12 , provide protection from ultraviolet radiation 13,14 , and stabilization of pigment proteins 15. In addition to the functions mentioned above, the involvement of carotenoids in cold adaptation was suspected 16-18. We assumed that carotenoids might have a similar function in regulating membrane fluidity as sterols such as cholesterol or ergosterol in eukaryotic cells. They increase membrane order with concurrent maintenance of lateral lipid motility, which results in a liquid-ordered membrane state 19. Similar mechanisms were previously described for the bacterial sterol-like hopanoids 20,21 and isoprenoid quinones 22. In vitro studies wit...

is a food pathogen capable of growing at a broad temperature range from 50°C to refrigerator temperatures. A key requirement for bacterial activity and growth at low temperatures is the ability to adjust the membrane lipid composition to maintain cytoplasmic membrane fluidity. In this study, we confirmed earlier findings that the extents of fatty acid profile adaptation differed between strains. We were able to demonstrate for isolates from food that growth rates at low temperatures and resistance to freeze-thaw stress were not impaired by a lower adaptive response of the fatty acid composition. This indicated the presence of a second adaptation mechanism besides temperature-regulated fatty acid synthesis. For strains that showed weaker adaptive responses in their fatty acid profiles to low growth temperature, we could demonstrate a significantly higher concentration of isoprenoid quinones. Three strains even showed a higher quinone concentration after growth at 6°C than at 37°C, which is contradictory to the reduced respiratory activity at lower growth temperatures. Analyses of the membrane fluidity by measuring generalized polarization and anisotropy revealed modulation of the transition phase. Strains with increased quinone concentrations showed an expanded membrane transition phase in contrast to strains with pronounced adaptations of fatty acid profiles. The correlation between quinone concentration and membrane transition phase expansion was confirmed by suppression of quinone synthesis. A reduced quinone concentration resulted in a narrower transition phase. Expansion of the phase transition zone by increasing the concentration of non-fatty acid membrane lipids is discussed as an additional mechanism improving adaptation to temperature shifts for strains. is a foodborne pathogen with an outstanding temperature range for growth. The ability for growth at temperatures close to the freezing point constitutes a serious contamination potential for cold stored food. The only known mechanism of the species for adaptation of membrane fluidity is modification of the membrane fatty acid composition. We were able to demonstrate that, at least for some strains, this adaptation mechanism is supported by regulation of the menaquinone concentration. The increase of this neutral membrane lipid is correlated with fluidization of the membrane under low-temperature conditions and therefore represents a fatty acid-independent mechanism for adaptation to low temperatures.

Five isolates from chilled food and refrigerator inner surfaces and closely related reference strains of the species Escherichia coli, Listeria monocytogenes, Staphylococcus xylosus, Bacillus cereus, Pedobacter nutrimenti, and Pedobacter panaciterrae were tested for the effect of growth temperature (30°C and 10°C) on biomass formation. Growth was monitored via optical density, and biomass formation was measured at the early stationary phase based on the following parameters in complex and defined media: viable cell count, total cell count, cell dry weight, whole-cell protein content, and cell morphology. According to the lack of growth at 1°C, all strains were assigned to the thermal class of mesophiles. Glucose and ammonium consumption related to cell yield were analyzed in defined media. Except for the protein content, temperature had a significant (t test, P < 0.05) effect on all biomass formation parameters for each strain. The results show a significant difference between the isolates and the related reference strains. Isolates achieved an increase in biomass production between 20% and 110% at the 10°C temperature, which is 15 to 25°C lower than their maximum growth rate temperatures. In contrast, reference strains showed a maximum increase of only about 25%, and some reference strains showed no increase or a decrease of approximately 25%. As expected, growth rates for all strains were higher at 30°C than at 10°C, while biomass production for isolates was higher at 10°C than at 30°C. In contrast, the reference strains showed similar growth yields at the two temperatures. This also demonstrates for mesophilic bacterial strains more efficient nutrient assimilation during growth at low temperatures. Until now, this characteristic was attributed only to psychrophilic microorganisms. IMPORTANCEFor several psychrophilic species, increased biomass formation was described at temperatures lower than optimum growth temperatures, which are defined by the highest growth rate. This work shows increased biomass formation at low growth temperatures for mesophilic isolates. A comparison with closely related reference strains from culture collections showed a significantly smaller increase or no increase in biomass formation. This indicates a loss of specific adaptive mechanisms (e.g., cold adaptation) for mesophiles during long-term cultivation. The increased biomass production for mesophiles under low-temperature conditions opens new avenues for a more efficient biotechnological transformation of nutrients to microbial biomass. These findings may also be important for risk assessment of cooled foods since risk potential is often correlated with the cell numbers present in food samples.T emperature has an obvious influence on the physiological functions and, consequently, on the growth and survival of bacteria (1, 2). Accordingly, bacteria have adapted to the specific temperature ranges of various environments, which can be summarized in different temperature classes.Cold-adapted microorganisms are classified as ps...

The diheme cytochromes c of the widespread TsdA family are bifunctional thiosulfate dehydrogenase/tetrathionate reductases. Here, biochemical information was collected about TsdA from the Epsilonproteobacterium Wolinella succinogenes (WsTsdA). The situation in W. succinogenes is unique since TsdA is closely associated with the unprecedented lipoprotein TsdC encoded immediately downstream of tsdA in the same direction of transcription. WsTsdA purified from Escherichia coli catalyzed both thiosulfate oxidation and tetrathionate reduction. After co-production of TsdC and WsTsdA in E. coli, TsdC was found to mediate membrane attachment of TsdA and to ensure its full catalytic activity. This effect was much stronger in the tetrathionate-reducing than in the thiosulfate-oxidizing direction. It is concluded that the TsdAC complex predominantly acts as a tetrathionate reductase in vivo.

In recent years, there has been a global trend towards a plant-based lifestyle. In the NuEva study, dietary self-reports of 258 participants following one of four diets (Western diet (WD), flexitarians (Flex), vegetarians (VG), and vegans (VN)) were related to fecal microbiome composition. Reduced consumption of animal products (VN < VG < Flex < WD) was associated with a decreased intake of energy (p < 0.05), and an increased intake of soluble and non-soluble dietary fibers (p < 0.05). We observed the lowest average microbiome diversity in vegans and the highest in WD. Compared to WD, VG (p < 0.05) and VN (p < 0.01) differed significantly in their bacterial composition. These data were related to dietary fiber intake. Furthermore, we identified 14 diet-specific biomarkers at the genus level by using LefSe analysis. Of these, 11 showed minimum or maximum counts in WD or VN. While the VN-specific species were inversely associated with cardiovascular risk factors, a positive association was detected for the WD-specific species. Identifying biomarkers for the diets on extreme ends of the spectrum (WD and VN) and their association with cardiovascular risk factors provides a solid evidence base highlighting the potential and the need for the development of personalized recommendations dependent on dietary patterns. Even so, the mechanisms underlying these diet-specific differences in microbiome composition cannot yet be clearly assessed. The elucidation of these associations will provide the basis for personalized nutritional recommendations based on the microbiome.