Effect of dietary fat upon aflatoxicosis in rats fed torula yeast containing diet

This study was designed to study the possible interrelationships between Torula yeast, vitamin E, and the dietary fat source on aflatoxin-induced tumors. Rats were fed Torula yeast-containing basal diets which included 1.7 ppm aflatoxin B₁ with either lard, corn oil or no fat, and with or without vitamin E supplements for 3 months. Thereafter, the respective diets without aflatoxin were fed for ca. 9 months. Animals receiving the vitamin E-deficient diets had a high mortality. Although the vitamin E-deficient, aflatoxin-treated rats had lower wt gains than did the vitamin E-deficient controls, they lived twice as long. In addition, regardless of the dietary fat source, the kidneys and adrenals of these vitamin E-deficient, aflatoxin-supplemented rats were found to be significantly heavier than the controls, and plasma cholesterol levels were elevated. Increased amounts of liver lipid were observed in response to aflatoxin in both corn oil-fed and fat-deficient rats. No such differences were observed in the responses of the vitamin E-supplemented groups to aflatoxin. On the corn oil diet, aflatoxin administration resulted in an increased deposition of polyunsaturated fatty acids in cholesteryl ester and phospholipid fractions in livers of vitamin E-deficient rats and the phospholipid fraction of vitamin E-sufficient rats. The vitamin E-deficient rats exhibited necrosis of the liver, which was alleviated when aflatoxin was included in the diet, and calcification of the kidneys, which was potentiated by the dietary aflatoxin. No tumors were observed in these animals. In animals maintained on vitamin E-sufficient diets for 1 year, growth was depressed as a result of aflatoxin administration with the greatest depression occurring in the group fed corn oil. Spleen wt were decreased in all groups given aflatoxin. However, there were no changes in either plasma or liver cholesterol or total liver lipids which could be attributed to aflatoxin administration. When aflatoxin was fed with lard, the cholesteryl ester, triglyceride, and free fatty acid fractions of plasma had decreased amounts of the C20:4 acid. In the cholesteryl ester fraction only, this change was accompanied by increased levels of C16:0, C18:0, and C18:1 acids. In the liver phospholipids, there were increased levels of mono- and polyunsaturated fatty acids and decreases in the saturated fatty acids. All of the animals receiving aflatoxin exhibited severe necrosis and tumor formation in the kidneys; the animals fed lard had the highest level of involvement and those in the fat-free group the least. Liver pathology was the least marked among the rats fed the fat-free diet. Since aflatoxin-induced tumors are rich in lipids, the fat-free diet may be protective to the animal.     

酵母菌固定化及对锶吸附的影响

酵母菌具有吸附富集重金属离子的能力.利用海藻酸钠对酿酒酵母菌进行固定,对固定化后的酿酒酵母菌吸附锶的能力做了对比研究.结果表明,海藻酸钠颗粒对锶有一定吸附作用,吸附速度较快,但是结合不稳定,未固定的酵母菌吸附较稳定,但是在低浓度时,吸附率不高;固定化酵母菌对锶的吸附分两个阶段,低质量浓度(10 mg/L)时,吸附率可达80%.在不同质量浓度梯度下(10～100 mg/L)吸附研究中发现,三者的吸附率均随浓度增大而减小,但吸附总量均有所增加.随培养时间延长,未固定与固定化酵母菌对锶的吸附率和吸附量趋于一致.结果表明海藻酸钠可以作为酵母菌活细胞生长固定化载体并表现出对低浓度Sr2+的较好吸附效果.     

Role of carrier material in encapsulation of yeast (Saccharomyces cerevisiae) by spray drying

ABSTRACT

Yeast was encapsulated using different carrier materials and their combinations to explore the possible synergistic effect of carrier material during encapsulation using spray drying. Freeze-drying was performed for comparison. The dried cell powders were analyzed for the quality aspects (morphology, flowability, and storage stability). The best results were observed, with a combination of whey protein and corn starch (cell survival: 82.37% and yield: 56%, w/w) with a shelf life of 6 months (with only 10% reduction in cell survival). The survival was found to be 40% without any carrier material, which decreased to less than 25% within 4 weeks.     

The increase in intracellular free-lysine levels of growing Baker's yeast (Saccharomyces cerevisiae) by sodium chloride

Fermentation by baker's yeast was carried out in medium containing exogenously added sodium chloride. The cell, intracellular free-lysine, glucose and ethanol concentrations were monitored at intervals of time during the fermentation. Sodium chloride (0.15–0.6M) stimulated the maximum intracellular free-lysine concentration of baker's yeast. The greatest stimulation was observed using 0.6M sodium chloride and was accompanied by a reduction in the rate of glucose uptake from the medium and a lowering of the ethanol concentration in the medium.     

Removal of heavy metals using a brewer's yeast strain of Saccharomyces cerevisiae: application to the treatment of real electroplating effluents containing multielements

BACKGROUND: Yeast cells have been recognized as an effective type of biomass for the treatment of wastewaters containing heavy metals. However, its capability to treat efficiently complex effluents loaded with several metals ions (Cr, Cu, Ni and Zn) has never been reported. The aim of this work was to study the feasibility of a hybrid technology, which combines chemical precipitation at pH 6.0 with a subsequent biotechnological‐based process (using heat‐killed cells of a flocculent brewing strain of Saccharomyces cerevisiae), to remove simultaneously several metals from real electroplating effluents. RESULTS: Two effluents containing Cu, Ni and Zn (effluent A) or Cr, Cu and Ni (effluent B) were treated. In both effluents, pH was adjusted to 6.0; in effluent B, Cr(VI) was previously reduced to Cr(III). Chemical speciation studies allowed defining the amount of biomass to be employed with a minimum number of batches. Subsequently to pH adjustment to 6.0, effluents were fully treated with a serial batch of biomass. After the third batch, metal concentrations were lowered to below the legal limits of discharge; removals ≥ 89, 91, 92 and 94% were attained for Ni, Cu, Cr and Zn, respectively. CONCLUSIONS: In the present work, the usefulness of using flocculent brewing yeast cells to treat complex industrial effluents loaded with several heavy metals was demonstrated. The hybrid process developed was shown to be an efficient alternative for the treatment of real electroplating effluents. Copyright © 2010 Society of Chemical Industry     

Co(II) and Ni(II) Concentration,and Co(II) Purification with Microbiological Collectors (Saccharomyces cerevisiae)

Abstract

Co(II) and Ni(II) can be concentrated quantitatively using a microbiological collector consisting of a Saccharomyces cerevisiae strain suspended in a glucose containing phosphate buffer. Optimal conditions for such accumulation as regards pH, time, and concentration have been studied. The influence of some complexing agents on the accumulation of a mixture of Co(II) and Ni(II) has also been investigated. By adapting the Saccharomyces cerevisiae strain to Co(II), separation of Co(II) from Ni(II) in dilute solution has been achieved.     

Effect of dietary calcium during pregnancy and lactation on zinc levels in blood and bone, in rats

The effect of dietary calcium (Ca) level on maternal zinc (Zn) nutritional status was studied. Female Wistar rats, weighing 250-350 g, were fed during pregnancy and lactation with an experimental diet containing/100 g different levels of calcium: 0.2 g (low calcium: LCa), 0.6 g (normal calcium: NCa) or 0.9 (high calcium: HCa). Maternal blood samples were drawn from the tail at delivery and at the end of lactation. Laboratory determinations were: Zn in whole blood (WB) at delivery and weaning; Zn (ZnF) and Ca (CaF) in the ashed femur at weaning. The results (mean +/- SEM) were: ZnWB (microgram/ml) at delivery and weaning: LCa: 8.73 +/- 1.05; 12.8 +/- 2.02; NCa: 3.49 +/- 0.19; 3.73 +/- 0.37; HCa: 3.21 +/- 0.19; 3.85 +/- 0.27. CaF (mg/100 mg): LCa: 19.2 +/- 0.8; NCa: 21.4 +/- 0.6; HCa: 20.4 +/- 1.1. ZnF (microgram/100 mg): LCa: 30.2 +/- 0.9; NCa: 24.1 +/- 0.3; HCa: 24.1 +/- 0.9. ZnWB was significantly higher in LCa (p < 0.0001) regarding NCa and Hca. ZnF showed an increase and CaF a decrease in LCa regarding NCa and HCa (p < 0.0001). There were no significant differences in ZnWB, ZnF and CaF between NCa and HCa These results show that: there was no detrimental effect when dietary Ca content was increased by 50% above the normal requirements of the rat.; low dietary Ca during pregnancy and lactation produced an increase of Zn utilization, reflected in maternal blood Zn and in ZnF content.     

Variations in mouse (Mus musculus) urinary volatiles during different periods of pregnancy and lactation

Mouse urine samples from different pregnancy and lactation periods were examined by capillary gas chromatography to assess variations in the volatile signals that may affect the endocrine function of other females. Statistically significant changes in the excretion of certain urinary volatiles were observed; from 26 readily quantifiable constituents, 14 appear to be under the endocrine control. These selected components, positively identified through mass spectrometry and retention data, and the synthetic standards are ketones, unsaturated alcohols, esters, and cyclic vinyl ethers.     

Production of an Active,Human Membrane Protein in Saccharomyces cerevisiae: Full-Length FICD

The human Fic domain-containing protein (FICD) is a type II endoplasmic reticulum (ER) membrane protein that is important for the maintenance of ER proteostasis. Structural and in vitro biochemical characterisation of FICD AMPylase and deAMPylase activity have been restricted to the soluble ER-luminal domain produced in Escherichia coli. Information about potentially important features, such as structural motifs, modulator binding sites or other regulatory elements, is therefore missing for the approximately 100 N-terminal residues including the transmembrane region of FICD. Expressing and purifying the required quantity and quality of membrane proteins is demanding because of the low yields and poor stability often observed. Here, we produce full-length FICD by combining a Saccharomyces cerevisiae-based platform with green fluorescent protein (GFP) tagging to optimise the conditions for expression, solubilisation and purification. We subsequently employ these conditions to purify milligram quantities of His-tagged FICD per litre of culture, and show that the purified, detergent-solubilised membrane protein is an active deAMPylating enzyme. Our work provides a straightforward methodology for producing not only full-length FICD, but also other membrane proteins in S. cerevisiae for structural and biochemical characterisation.     

Tryptophan Derivatives by Saccharomyces cerevisiae EC1118: Evaluation,Optimization, and Production in a Soybean-Based Medium

Given the pharmacological properti es and the potential role of kynurenic acid (KYNA) in human physiology and the pleiotropic activity of the neurohormone melatonin (MEL) involved in physiological and immunological functions and as regulator of antioxidant enzymes, this study aimed at evaluating the capability of Saccharomyces cerevisiae EC1118 to release tryptophan derivatives (dTRPs) from the kynurenine (KYN) and melatonin pathways. The setting up of the spectroscopic and chromatographic conditions for the quantification of the dTRPs in LC-MS/MS system, the optimization of dTRPs’ production in fermentative and whole-cell biotransformation approaches and the production of dTRPs in a soybean-based cultural medium naturally enriched in tryptophan, as a case of study, were included in the experimental plan. Variable amounts of dTRPs, with a prevalence of metabolites of the KYN pathway, were detected. The LC-MS/MS analysis showed that the compound synthesized at highest concentration is KYNA that reached 9.146 ± 0.585 mg/L in fermentation trials in a chemically defined medium at 400 mg/L TRP. Further experiments in a soybean-based medium confirm KYNA as the main dTRPs, whereas the other dTRPs reached very lower concentrations. While detectable quantities of melatonin were never observed, two MEL isomers were successfully measured in laboratory media.     

Effect of sterculic acid upon aflatoxicosis in rats fed diets containing saturated and unsaturated fat

Investigations on trout have shown that the cyclopropenoid fatty acids, which occur naturally in small amounts in unrefined cottonseed oil, may act as powerful cocarcinogens when fed in conjunction with aflatoxin. Attempts at confirming these findings in mammals, i.e. rats, have been inconclusive. In this study, the effects of sterculic acid and aflatoxin upon lipid metabolism and tumor formation in male rats have been examined using basal diets containing either saturated or unsaturated fat to which the following additions were made: (A) basal diet (no supplements); (B) aflatoxin B₁ at 1.7 ppm; (C) sterculic acid at 210 ppm; and (D) aflatoxin B₁ at 1.7 ppm, plus sterculic acid at 210 ppm. The rats consumed these diets for 3 months and, thereafter, were fed the unsupplemented basal diet until sacrifice 9 months later. Growth was depressed in rats in groups B, C, and D, but no synergistic inhibition was observed, regardless of the fat source. Liver wt doubled in response to aflatoxin; however, only when the diet contained unsaturated fat did sterculic acid, in combination with aflatoxin, exaggerate the increase in liver wt (a reflection of the more severe liver pathology observed in these rats). In the animals fed the saturated fat diet, aflatoxin administration to animals fed the control or sterculic acid supplemented diets resulted in marked increases in plasma cholesterol levels; the unsaturated fat diets, supplemented with aflatoxin, evoked a slight increase in plasma cholesterol content which was nullified by sterculic acid supplementation.     

Differential temperature regulation of three sunflower microsomal oleate desaturase (FAD2) isoforms overexpressed in Saccharomyces cerevisiae

The oxygen‐independent temperature regulation of three sunflower microsomal oleate desaturase (FAD2) isoforms has been investigated using Saccharomyces cerevisiae cells expressing each FAD2 gene. Yeast cells transformed with the FAD2‐1 gene showed the highest percentage of dienoic acids when they were grown at 10–15 °C. In contrast, the maximal level of dienoic acids for S. cerevisiae cells expressing the FAD2‐2 and FAD2‐3 genes were obtained at 30 and 35 °C, respectively. Temperature shifts in the phase of exponential growth, from 30 to 15 °C or from 15 to 30 °C, produced changes in the final percentage of dienoic acids, mostly in yeast cells transformed with the FAD2‐1 gene, to reach the content corresponding to the new temperature. Low temperature (15 °C) increased the amount of neutral lipids in all transformed yeast cells, mainly because it favored triacylglycerol accumulation. In addition, the FAD2‐expressing yeast cells showed a higher polar lipid content than those transformed with the empty vector. Dienoic acids were present in all lipids, although high temperature (30 °C) favored their accumulation in neutral lipids. As the main conclusion, the low thermal stability observed for the major and seed specific isoform (FAD2‐1) is the key factor controlling the direct temperature regulation in sunflower seeds.     

The effect of clofibrate on heart and plasma lipids in rats fed a diet containing rapeseed oil

The effect of clofibrate on heart and plasma lipids in rats fed a diet containing 30% of the calories as peanut oil (PO) or rapeseed oil (RSO) (42.7% erucic acid and 0.5% eicosenoic acid) was studied. A decrease of erucic acid content to one-third and concomitant increase in the content of 18∶1, 16∶1 and 16∶0 fatty acids in plasma triacylglycerols were observed after administration of clofibrate to rats fed the RSO-diet. It is suggested that these changes reflect the increased capacity of the liver to chainshorten very long chain length fatty acids. The extent of lipidosis in the heart of rats fed the RSO-diet was decreased by 50% by clofibrate. However, the concentration of erucic acid in heart triacylglycerols decreased much less (30%) than the concentration of all other fatty acids (50–65%). It is concluded that the clofibrate administration increased the oxidative capacity of the heart mitochondria and that the heart cell does not have an efficient system to handle very long chain length monounsaturated fatty acids as does the liver.     

Quantitative and qualitative effects of larval diet on male scent secretions ofEstigmene acrea,Phragmatobia foliginosa,andPyrrharctia isabella (Lepidoptera: Arctiidae)

In feeding experiments with insects reared in the laboratory, the presence of the dihydropyrrolizines hydroxydanaidal and danaidal in the male scent organs (coremata) of the arctiids,Estigmene acrea (Drury),Phragmatobia fuliginosa (L.), andPyrrharctia isabella (J.E. Smith), was shown to depend on the presence of a source of pyrrolizidine alkaloids (PAs) in the larval diet.Phragmatobia males given an artificial diet supplemented with the powdered roots of the PA-containing plantSymphytum officinale L. (comfrey) produced more hydroxydanaidal than danaidal, whereas males given an artificial diet supplemented with dried whole plants of another PA-containing species,Senecio vulgaris L., produced more danaidal than hydroxydanaidal.Pyrrharctia males produced hydroxydanaidal with little if any danaidal, whether the source of PAs was comfrey orS. vulgaris. A behavioral bioassay showed that the coremata of PA-deniedPyrrharctia male progeny of PA-denied parents were pheromonally inactive, whereas those of PA-denied male progeny of PA-supplied parents (male and/or female) were often active. This indicates that a small amount of pheromone is made from PAs transferred from the female to her eggs and that males effect copulatory transfers of PAs that are, in turn, passed to the eggs by the mated female. Field observations ofPhragmatobia andPyrrharctia larvae feeding on sources of PAs were reported. The PA monocrotaline was shown to be a feeding stimulant forPyrrharctia larvae.     

Entrapment of Saccharomyces cerevisiae cells in u.v. crosslinked hydroxyethylcellulose/poly(ethylene oxide) double-layered gels

Double-layered gels, consisting of hydroxyethylcellulose cryogel core and poly(ethylene oxide) hydrogel shell, were synthesized with u.v. irradiation, using the same photoinitiator, (4-benzoylbenzyl) trimethylammoniumchloride (BBTMAC) for the both layers. The gels were characterized by measuring their rheological parameters, gel fraction yield, the degree of equilibrium swelling and diffusion coefficient. The diffusion coefficients for glucose and ethanol through the hydroxyethylcellulose cryogel were 3.9 × 10^?6 cm²/s and 0.97 × 10^?5 cm²/s, respectively. The applicability of these double-layered gels as carriers for immobilization was investigated by entrapment of Saccharomyces cerevisiae cells. The immobilization efficiency and cell retention were determined in batch fermentation for ethanol production from glucose. The operational stability of the gels was evaluated in batch fermentation with three consecutive runs. The ethanol yield was in the range from 60% to 77% of the theoretical yield.     

A Heavy Metal-Associated Protein (AcHMA1) from the Halophyte,Atriplex canescens (Pursh) Nutt., Confers Tolerance to Iron and Other Abiotic Stresses When Expressed in Saccharomyces cerevisiae

Many heavy metals are essential for metabolic processes, but are toxic at elevated levels. Metal tolerance proteins provide resistance to this toxicity. In this study, we identified and characterized a heavy metal-associated protein, AcHMA1, from the halophyte, Atriplex canescens. Sequence analysis has revealed that AcHMA1 contains two heavy metal binding domains. Treatments with metals (Fe, Cu, Ni, Cd or Pb), PEG6000 and NaHCO₃ highly induced AcHMA1 expression in A. canescens, whereas NaCl and low temperature decreased its expression. The role of AcHMA1 in metal stress tolerance was examined using a yeast expression system. Expression of the AcHMA1 gene significantly increased the ability of yeast cells to adapt to and recover from exposure to excess iron. AcHMA1 expression also provided salt, alkaline, osmotic and oxidant stress tolerance in yeast cells. Finally, subcellular localization of an AcHMA1/GFP fusion protein expressed in tobacco cells showed that AcHMA1 was localized in the plasma membrane. Thus, our results suggest that AcHMA1 encodes a membrane-localized metal tolerance protein that mediates the detoxification of iron in eukaryotes. Furthermore, AcHMA1 also participates in the response to abiotic stress.