Study of selenium enrichment metabolomics in Bacillus subtilis BSN313 via transcriptome analysis.

In this study, the transcriptome analysis was practiced to identify potential genes of probiotic Bacillus subtilis BSN313 involved in selenium (Se) enrichment metabolism. The transcriptomic variation of the strain was deliberated in presence of three different sodium selenite concentrations (0, 3, and 20 µg/mL). The samples were taken at 1 and 13 h subsequent to inoculation of selenite and gene expression profiles in Se metabolism were analyzed through RNA sequencing. The gene expression levels of the pre log phase were lower than the stationary phase. It is because, the bacteria has maximum grown with high concentration of Se (enriched with organic Se), at stationary phase. Bacterial culture containing 3 µg/mL concentration of inorganic Se (sodium selenite) has shown highest gene expression as compared to no or high concentration of Se. This concentration (3 µg/mL) of sodium selenite (as Se) in the medium promoted the upregulation of thioredoxin reductase expression, whereas its higher Se concentration inhibited the formation of selenomethionine (SeMet). The result of 5 L bioreactor fermentation showed that SeMet was also detected in the fermentation supernatant as the growth entered in the late stationary phase and reached up to 857.3 ng/mL. The overall intracellular SeMet enriched content in BSN313 was extended up to 23.4 µg/g dry cell weight. The other two selenoamino acids (Se-AAs), methyl-selenocysteine, and selenocysteine were hardly detected in medium supernatant. From this study, it was concluded that SeMet was the highest content of organic Se byproduct biosynthesized by B. subtilis BSN313 strain in Se-enriched medium during stationary phase. Thus, B. subtilis BSN313 can be considered a commercial probiotic strain that can be used in the food and pharmaceutical industries. This is because it can meet the commercial demand for Se-AAs (SeMet) in both industries.

Single-cell ICP-MS for evaluating the Se-protective effect against MeHg+-induced neurotoxicity in human neuroblastoma cell line (SH-SY5Y).

The protective effect of selenium (Se) against Hg-induced neurotoxicity has been widely investigated; however, the mechanisms behind this interaction have not been fully elucidated yet. In the current work, the role of Se against MeHg+-induced cytotoxicity in the human neuroblastoma cell line (SH-SY5Y) is reported for the first time by tracking Hg uptake and accumulation at the single-cell level by inductively coupled plasma-mass spectrometry in single-cell mode (SC-ICP-MS). The influence of different Se species (SeMet, SeMeSeCys, citrate-SeNPs, and chitosan-SeNPs) on MeHg+ cytotoxicity was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays. SeMet and SeMeSeCys exhibited protective effects against MeHg+-induced cell death, particularly at high MeHg+ concentrations (LC50). In addition, chitosan-SeNPs showed greater protection compared to citrate-SeNPs when co-exposed with MeHg+. Interestingly, SC-ICP-MS unveiled the heterogeneous distribution of Hg uptake by SH-SY5Y cells. Co-exposure of SeMet and SeMeSeCys with MeHg+ led to a reduction of the amount of Hg accumulated per individual cell, which decreased the maximum level of Hg per cell by half (from 60 fg Hg cell-1 to 30 fg Hg cell-1) when SeMet was present, along with a decrease in the percentage of cells that accumulated the highest quantity of MeHg+. All these data corroborate the protective role of Se against Hg toxicity at the cellular level.

A reliable method of high performance liquid chromatography coupled with inductively coupled plasma mass spectrometry for determining selenoamino acids in selenoproteins from Lactococcus lactis.

A reliable method for simultaneous determination of four organic selenium species by HPLC-ICP-MS was developed and implemented in determining organic selenoamino acids (Se-AAs) in selenoproteins from Lactococcus lactis (L. lactis) NZ9000. The method consisted of liberating Se-AAs from selenoproteins using ultrasound-assisted protease hydrolysis, and quantitatively detecting Se-AA speciations by HPLC-ICP-MS. After optimizations of proteolysis conditions, chromatographic conditions and determination conditions, the established method could efficiently separate the four Se-AAs, including SeCys, SeCys2, SeMeCys and SeMet within 10 min. It presented high sensitivity with the limits of detection and quantitation in the range of 0.197∼0.240 µg∙L-1 and 0.788∼0.960 µg∙L-1, respectively, good repeatability with a relative standard deviation (RSD) of less than 5%, and good recovery in the desired floating range of 90%∼105%, verifying the good accuracy. The method successfully detected four selenium species in the purified glutathione peroxidase (LlGPx) overexpressed in L. lactis NZ9000, SeCys (0.9716∼1.6784 µg∙g-1), SeCys2 (1.0695∼1.2124 µg∙g-1), SeMeCys (0.7288∼0.7984 µg∙g-1) and SeMet (1.0058∼1.9571 µg∙g-1), accounting for up to 80.14% of total selenium. There was no difference of order of magnitude in the four Se-AAs, indirectly indicating the random incorporation of selenium into selenoprotein LlGPx in L. lactis NZ9000. This work throws new light on the identification and biosynthesis of organic selenium species in selenoproteins and selenium-riched organisms like L. lactis.

Is selenium beneficial or detrimental to earthworm? Growth and metabolism responses of Eisenia Fetida to Na2SeO3 exposure.

Se unevenly distributed in soils due to variations of geology and anthropogenic input, which results in different effects on earthworms. The effects of Se were characterized by analyzing the growth and metabolism responses of earthworms after exposure to three different concentrations of Na2SeO3. The results showed that except the possible growth promotion at 5 mg/kg, low and middle-level exposure to Na2SeO3 (0.3-10 mg/kg) did not significantly affect the growth of earthworms. While a significant inhibition effect on growth was observed in the high-level exposure group (30-70 mg/kg). There was an inflection point for Se performing promotion to inhibition effects on earthworm growth. To investigate the metabolic response of earthworms, a novel HPLC-ESI-MS (High Performance Liquid Chromatography-Electrospray Ionization-Mass Spectrometry) method was used to determine sensitive biomarkers. Selenium exposure significantly altered the metabolism of seven essential amino acids, namely tyrosine, leucine, phenylalanine, valine, alanine, glycine, and lysine, and two selenoamino acids, namely selenomethionine and methylselenocysteine. The overall metabolism level of earthworms was not affected at low exposure concentrations, but was affected at medium and high exposure concentrations. The metabolic pathways that integrated the selenocompound metabolism and the tricarboxylic acid cycle from the perspective of energy supply and demand were affected by Na2SeO3 exposure. The derived reactive oxygen species at high exposure concentrations were probably the reason for the growth inhibition effect of Se on earthworms. This study provides biochemical insights into the effects of Na2SeO3 on earthworms and suggests that an Se concentration of about 2.3 mg/kg is appropriate for soil organism health.

Effect of boiling and frying on the selenium content, speciation, and in vitro bioaccessibility of selenium-biofortified potato (Solanum tuberosum L.).

Selenium-enriched potato is a good supplement for selenium-deficient populations. This study evaluated the influence of two most common cooking methods, including boiling and frying, on selenium content, speciation, and in vitro bioaccessibility of selenium-biofortified potato tubers. After foliar application of 200 µg/mL sodium selenite, potato tubers with 1.33 µg Se/g were obtained. Peeling resulted in 53.4%-69.9% loss of selenium in tubers. The total selenium content decreased by approximately 43.3% after boiling, among which up to 38.5% of the lost selenium is found in the boiling water. Nearly 31.7% of selenium was lost via volatilization during frying. Both cooking methods significantly enhanced the bioaccessibility of Se(IV) in tubers. Whereas SeMeCys became less bioaccessible after boiling. SeMet and SeCys2 in fired tubers were not accessible after digestion. This study suggested that boiling is more appropriate for cooking selenium-enriched potatoes.

Selenium accumulation, speciation, and its effect on nutritive value of Flammulina velutipes (Golden needle mushroom).

Flammulina velutipes is one of the most popular edible mushrooms worldwide. A selenium-biofortification method for its fruiting body was developed using selenite. This study investigated the selenium content, distribution, speciation and the effect of selenium on mushroom growth, nutritive value, and mineral accumulation. Results showed that F. velutipes accumulated nearly 108 µg/g of organic selenium under treatment with 20 µg/g selenite, which accounts for over 97% of total selenium. Most (60-74%) of selenium combined with the protein fraction, whereas 15-21% combined with the polysaccharide fraction. Selenomethionine (56.8%), selenocysteine (22.8%), and methylselenocysteine (17.3%) were the main organic selenium compounds in the fruiting body. Selenium biofortification increased the biomass yield of fruiting body and elevated the content of polysaccharides, proteins, total amino acids, essential amino acids, and several minerals, including iron, calcium, and copper. F. velutipes might become a suitable selenium supplement.

Accumulation and speciation of selenium in biofortified vegetables grown under high boron and saline field conditions.

Selenium (Se) biofortification, as an agronomic-based strategy, is utilized to produce Se-enriched food products for increasing Se intake in inhabitants in Se-deficient regions. This strategy can be accomplished by soil and foliar application of Se or by growing crops in soils naturally high in Se. In this study, different cruciferous vegetables were field-grown in high boron (B) and saline soils of central California containing naturally high levels of Se. We investigated whether Se biofortification occurs in salt- and B-tolerant vegetables grown in poor-quality soil. The uptake of Se and other elements occurred in all vegetables. In plant tissues, Se speciation analyses showed greatest percentages of Se-containing compounds were contained in organic Se forms (monomethylated) and as selenate in the inorganic Se forms. Selenium-enriched vegetables produced from saline soils high in B and Se can be a natural source of Se-biofortified food that can be consumed as bioactive food products.

In vitro bioaccessibility of selenoamino acids from selenium (Se)-enriched Chlorella vulgaris biomass in comparison to selenized yeast; a Se-enriched food supplement; and Se-rich foods.

Selenium (Se) is an indispensable microelement in our diet and health issues resulting from deficiencies are well documented. Se-containing food supplements are available on the market including Se-enriched Chlorella vulgaris (Se-Chlorella) which accumulates Se in the form of Se-amino acids (Se-AAs). Despite its popular uses, data about the bioaccessibility of Se-AAs from Se-Chlorella are completely missing. In the present study, gastrointestinal digestion times were optimized and the in vitro bioaccessibility of Se-AAs in Se-Chlorella, Se-yeast, a commercially available Se-enriched food supplement (Se-supplement) and Se rich foods (Se-foods) were compared. Higher bioaccessibility was found in Se-Chlorella (∼49%) as compared to Se-yeast (∼21%), Se-supplement (∼32%) and Se-foods. The methods used in production of Se-Chlorella biomass were also investigated. We found that disintegration increased bioaccessibility whereas the drying process had no effect. Similarly, temperature treatment by microwave oven also increased bioaccessibility whereas boiling water did not.

Identification of trace levels of selenomethionine and related organic selenium species in high-ionic-strength waters.

A new anion-exchange chromatographic separation method was used for the simultaneous speciation analysis of selenoamino acids and the more ubiquitous inorganic selenium oxyanions, selenite and selenate. For quantification, this separation was coupled to inductively coupled plasma-mass spectrometry to achieve an instrumental detection limit of 5 ng Se L(-1) for all species. This chromatographic method was also coupled to electrospray tandem mass spectrometry to observe the negative ion mode fragmentation of selenomethionine and one of its oxidation products. Low detection limits were achieved, which were similar to those obtained using inductively coupled plasma-mass spectrometry. An extensive preconcentration and cleanup procedure using cation-exchange solid-phase extraction was developed for the identification and quantification of trace levels of selenomethionine in environmental samples. Preconcentration factors of up to five were observed for selenomethionine, which in addition to the removal of high concentrations of sulphate and chloride from industrial process waters, allowed for an unambiguous analysis that would have been impossible otherwise. Following these methods, selenomethionine was identified at an original concentration of 3.2 ng Se L(-1) in samples of effluent collected at a coal-fired power plant's biological remediation site. It is the first time that this species has been identified in the environment, outside of a biological entity. Additionally, oxidation products of selenomethionine were identified in river water and laboratory algal culture samples. High-resolution mass spectrometry was employed to postulate the chemical structures of these species.