Intracellular glycerol accumulation in light-limited Dunaliella tertiolecta culture is determined by partitioning of glycerol across the cell membrane.

Dunaliella accumulates intracellular glycerol to counterbalance the extracellular salinity. In N-limited chemostat cultures of D. tertiolecta, total glycerol production (sum of intracellular and extracellular) and intracellular glycerol content were proportional to the salinity of the culture medium. In the light-limited D. tertiolecta culture, total glycerol output (sum of intracellular and extracellular) was relatively constant at different salinities (0.5 and 2.0 M), while the intracellular glycerol content was proportional to the culture medium salinity, that is, the cells released less glycerol into the culture medium, rather than de novo synthesis of glycerol at high culture medium salinity. The study implies different regulatory mechanisms in the accumulation of intracellular glycerol in N-limited and light-limited D. tertiolecta in response to salinity.

Occurrence of glycerol uptake in Dunaliella tertiolecta under hyperosmotic stress.

The unicellular halotolerant green alga species Dunaliella are able to proliferate in extremely varied salinities by synthesizing intracellular glycerol and adjusting the cell shape and volume. However, some marine Dunaliella species such as Dunaliella tertiolecta are not able to regulate cell volume as an immediate response to counter external osmotic shock. Here we report that a rapid shock-response mechanism is present in Dunaliella tertiolecta, involving uptake of exogenous glycerol in response to hyperosmotic shock without changing cell volume, and this glycerol uptake activity is associated with the Dunaliella tertiolecta glycerol uptake protein 1 (DtGUP1) gene, which belongs to the membrane-bound O-acyltransferase. The mutant DtGUP1-E, in which the DtGUP1 gene is silenced, displayed an inability to take up glycerol from the medium and showed cell death under hyperosmotic shock. To our knowledge, this is the first time a gene product has been reported in Dunaliella tertiolecta that is involved in glycerol uptake activity under hyperosmotic stress.

Expression of the Chlamydomonas reinhardtii sedoheptulose-1,7-bisphosphatase in Dunaliella bardawil leads to enhanced photosynthesis and increased glycerol production.

Bioengineering of photoautotrophic microalgae into CO(2) scrubbers and producers of value-added metabolites is an appealing approach in low-carbon economy. A strategy for microalgal bioengineering is to enhance the photosynthetic carbon assimilation through genetically modifying the photosynthetic pathways. The halotolerant microalgae Dunaliella possess a unique osmoregulatory mechanism, which accumulates intracellular glycerol in response to extracellular hyperosmotic stresses. In our study, the Calvin cycle enzyme sedoheptulose 1,7-bisphosphatase from Chlamydomonas reinhardtii (CrSBPase) was transformed into Dunaliella bardawil, and the transformant CrSBP showed improved photosynthetic performance along with increased total organic carbon content and the osmoticum glycerol production. The results demonstrate that the potential of photosynthetic microalgae as CO(2) removers could be enhanced through modifying the photosynthetic carbon reduction cycle, with glycerol as the carbon sink.

Effect of tea phenolics and their aromatic fecal bacterial metabolites on intestinal microbiota.

Tea is rich in polyphenols and other phenolics that have been widely reported to have beneficial health effects. However, dietary polyphenols are not completely absorbed from the gastrointestinal tract and are metabolized by the gut microflora so that they and their metabolites may accumulate to exert physiological effects. In this study, we investigated the influence of the phenolic components of a tea extract and their aromatic metabolites upon bacterial growth. Fecal homogenates containing bacteria significantly catalyzed tea phenolics, including epicatechin, catechin, 3-O-methyl gallic acid, gallic acid and caffeic acid to generate aromatic metabolites dependent on bacterial species. Different strains of intestinal bacteria had varying degrees of growth sensitivity to tea phenolics and metabolites. Growth of certain pathogenic bacteria such as Clostridium perfringens, Clostridium difficile and Bacteroides spp. was significantly repressed by tea phenolics and their derivatives, while commensal anaerobes like Clostridium spp., Bifidobacterium spp. and probiotics such as Lactobacillus sp. were less severely affected. This indicates that tea phenolics exert significant effects on the intestinal environment by modulation of the intestinal bacterial population, probably by acting as metabolic prebiotics. Our observations provide further evidence for the importance of colonic bacteria in the metabolism, absorption and potential activity of phenolics in human health and disease. The bioactivity of different phenolics may play an important role in the maintenance of gastrointestinal health.

Human fecal water modifies adhesion of intestinal bacteria to Caco-2 cells.

The aqueous phase of feces (fecal water) has been suggested to mediate the effects of diet on colon carcinogenesis. We determined whether human fecal water samples, of varying genotoxic potential, had the capacity to alter adhesion of intestinal bacteria to intestinal (Caco-2) cells. Genotoxicity of fecal water samples was measured using the single-cell gel electrophoresis assay ("comet" assay), and bacterial adhesion was measured using a well-established model system. Fecal water genotoxicity was found to correlate positively with inhibition of adhesion of Escherichia coli strains, Salmonella species, and Enterococcus faecium to Caco-2 cells. The presence of fecal water samples did not interfere with adhesion of Bacteroides and Lactobacillus species. Inhibition of adhesion by fecal water was not due to cytotoxicity to Caco-2 cells as cytotoxicities of most fecal water samples were similar, nor was the inhibitory effect due to bacteriotoxicity as toxicity of fecal waters in the 10 strains of bacteria studied was not detected. Results indicate that components in fecal water may alter adhesion of intestinal bacteria to intestinal cell surfaces and that this effect may be correlated to the genotoxic potential of fecal water. This may have consequences for dietary effects on colon carcinogenesis.