Effect of source-separated urine storage on estrogenic activity detected using bioluminescent yeast Saccharomyces cerevisiae.

The objective was to demonstrate that a microbial whole cell biosensor, bioluminescent yeast, Saccharomyces cerevisiae (BMAEREluc/ERα) can be applied to detect overall estrogenic activity from fresh and stored human urine. The use of source-separated urine in agriculture removes a human originated estrogen source from wastewater influents, subsequently enabling nutrient recycling. Estrogenic activity in urine should be diminished prior to urine usage in agriculture in order to prevent its migration to soil. A storage period of 6 months is required for hygienic reasons; therefore, estrogenic activity monitoring is of interest. The method measured cumulative female hormone-like activity. Calibration curves were prepared for estrone, 17ß-estradiol, 17α- ethinylestradiol and estriol. Estrogen concentrations of 0.29-29,640 µg L(-1) were detectable while limit of detection corresponded to 0.28-35 µg L(-1) of estrogens. The yeast sensor responded well to fresh and stored urine and gave high signals corresponding to 0.38-3,804 µg L(-1) of estrogens in different urine samples. Estrogenic activity decreased during storage, but was still higher than in fresh urine implying insufficient storage length. The biosensor was suitable for monitoring hormonal activity in urine and can be used in screening anthropogenic estrogen-like compounds interacting with the receptor.

Draft Genome Sequence of the Hydrogen- and Ethanol-Producing Anaerobic Alkalithermophilic Bacterium Caloramator celer.

Caloramator celer strain JW/YL-NZ35 is a Gram-positive thermophilic, alkalitolerant, and strictly anaerobic bacterium capable of producing hydrogen and ethanol under extreme conditions. The draft genome sequence presented here will provide valuable information to further explore the physiology of this species and its potential for biofuel production.

Genome Sequence of Halanaerobium saccharolyticum subsp. saccharolyticum Strain DSM 6643T, a Halophilic Hydrogen-Producing Bacterium.

Halanaerobium saccharolyticum is a halophilic anaerobic fermentative bacterium capable of producing hydrogen, a potential future energy carrier molecule. The high-quality draft genome of H. saccharolyticum subsp. saccharolyticum strain DSM 6643(T) consists of 24 contigs for 2,873,865 bp with a G+C content of 32.3%.

1,3-Propanediol production and tolerance of a halophilic fermentative bacterium, Halanaerobium saccharolyticum subsp. saccharolyticum.

1,3-Propanediol (1,3-PD) is widely used in polymer industry in production of polyethers, polyesters and polyurethanes. In this article, a study on 1,3-PD production and tolerance of Halanaerobium saccharolyticum subsp. saccharolyticum is presented. 1,3-PD production was optimized for temperature, vitamin B(12) and acetate concentration. The highest 1,3-PD concentrations and yields (0.6 mol/mol glycerol) were obtained at vitamin B12 concentration 64 µg/l and an inverse correlation between 1,3-PD and hydrogen production was observed with varying vitamin B12 concentrations. In the studied temperature range and initial acetate concentrations up to 10 g/l, no significant variations were observed in 1,3-PD production. High initial acetate (29-58 g/l) was observed to cause slight decrease in 1,3-PD concentrations produced but no effects on 1,3-PD yields (mol/mol glycerol). Initial 1,3-PD concentrations inhibited the growth of H. saccharolyticum subsp. saccharolyticum. When initial 1,3-PD concentration was raised from 1g/l to 57 g/l, a decrease of 12% to 75%, respectively, in the highest optical density was observed.

Halophilic anaerobic fermentative bacteria.

In hypersaline environments bacteria are exposed to a high osmotic pressure caused by the surrounding high salt concentrations. Halophilic microorganisms have specific strategies for balancing the osmotic pressure and surviving in these extreme conditions. Halophilic fermentative bacteria form taxonomically and phylogenetically a coherent group mainly belonging to the order Halanaerobiales. In this review, halophilic anaerobic fermentative bacteria in terms of taxonomy and phylogeny, special characteristics, survival strategies, and potential for biotechnological applications in a wide variety of branches, such as production of hydrogen, are discussed.

Hydrogen production from glycerol using halophilic fermentative bacteria.

Glycerol-based hydrogen production by the halophilic bacteria Halanaerobium saccharolyticum subspecies saccharolyticum and senegalensis was studied as batch experiments. The main metabolites of glycerol fermentation of both strains were hydrogen, carbon dioxide, and acetate. Subspecies saccharolyticum also produced 1,3-propanediol (1,3-PD), butyrate, and ethanol. The highest hydrogen yields were achieved with 2.5g/l glycerol and 150g/l salt at pH 7.4 (subsp. saccharolyticum, yield 0.6mol/mol glycerol) and at pH 7.0 (subsp. senegalensis, yield 1.6mol/mol glycerol). The hydrogen yield of subsp. senegalensis has potential for practical applications after scale-up and bioprocess optimizations and metabolic engineering after genome-wide sequencing could be applied to improve the yield of subsp. saccharolyticum.

A novel biosensor for the detection of zearalenone family mycotoxins in milk.

In this study, a method for detecting estrogenic mycotoxin residues in milk was developed utilizing bioluminescent whole-cell biosensors. Milk products of various compositions were spiked with the estrogenic mycotoxins zearalenone and its metabolites zearalanone, alpha-zearalanol, beta-zearalanol, alpha-zearalenol and beta-zearalenol. The estrogenic response was detected by a whole-cell biosensor based on a genetically modified Saccharomyces cerevisiae strain that in the presence of an estrogenic compound produces firefly luciferase-enzyme and further light emission within a system provided with D-luciferin substrate. The results show that the yeast sensor reacts to mycotoxins with typical sigmoidal response at nanomolar concentrations. The response differs in different milk products with regard to the fat content of the milk. Due to short assay time of less than 3h and automation the approach can be used as a bioavailability and activity screening method prior to more detailed chemical analysis.

Real-time Monitoring of Non-specific Toxicity Using a Saccharomyces cerevisiae Reporter System.

Baker's yeast, Saccharomyces cerevisiae, is the simplest and most well-known representative of eukaryotic cells and thus a convenient model organism for evaluating toxic effects in human cells and tissues. Yeast cell sensors are easy to maintain with short generation times, which makes the analytical method of assessing antifungal toxicity cheap and less-time consuming. In this work, the toxicity of test compounds was assessed in bioassays based on bioluminescence inhibition and on traditional growth inhibition on agar plates. The model organism in both tests was a modified S. cerevisiae sensor strain that produces light when provided with D-luciferin in an insect luciferase reporter gene activity assay. The bioluminescence assay showed toxic effects for yeast cell sensor of 5,6-benzo-flavone, rapamycin, nystatin and cycloheximide at concentrations of nM to µM. In addition, arsenic compounds, cadmium chloride, copper sulfate and lead acetate were shown to be potent non-specific inhibitors of the reporter organism described here. The results from a yeast agar diffusion assay correlated with the bioluminescence assay results.