Glutathione transferases(GSTs) play an important role in the detoxification of xenobiotic/endobiotic toxic compounds. The α-, π-, and/l-classes of cytosolic GSTs have been studied extensively, while Gtt2 from Sacc...Glutathione transferases(GSTs) play an important role in the detoxification of xenobiotic/endobiotic toxic compounds. The α-, π-, and/l-classes of cytosolic GSTs have been studied extensively, while Gtt2 from Saccharo- myces cerevisiae, a novel atypical GST, is still poorly understood. In the present study, we investigated the gluta- thione(GSH) activation mechanism of Gtt2 using the density functional theory(DFT) with the hybrid functional B3LYP. The computational results show that a water molecule could assist a proton transfer between the GSH thiol and the N atom of His133. The energy barrier of proton transfer is 46.0 kJ/mol. The GSH activation mechanism and the characteristics of active site are different from those of classic cytosolic GSTs.展开更多
[Objective] The aim of this study was to screen Saccharomyces for glutathione over-production. [Method] Ethionine-resistant mutants were obtained through UV mutagenesis and rational screening. [Result] A high GSH-prod...[Objective] The aim of this study was to screen Saccharomyces for glutathione over-production. [Method] Ethionine-resistant mutants were obtained through UV mutagenesis and rational screening. [Result] A high GSH-producing strain HSJB1 was isolated from soil, and the biomass for this strain by flask shaking fermentation was 3.87 g/L while the GSH yield was 91.87 mg/L. According to the morphological, physiological and biochemical characteristics of cells, this strain was primarily identified as Saccharomyces cerevisiae. An ethionine-resistant mutant YBS77 was obtained through UV mutagenesis of the original strain HSJB1, and the biomass for this strain by flask shaking fermentation was 7.60 g dry cell weight/L while the GSH yield was 211.96 mg/L. [Conclusion] The biomass of the mutant obtained by breeding is increased by 96.38% than that of the original strain, and the GSH yield of the mutant obtained by breeding is increased by 130.72% than that from the original strain, which indicates that the breeding method is feasible.展开更多
To evaluate the effects of high fat food consisted of tallow (19% of total diets) and cholesterol (2%) on modification of erythrocyte phospholipids, fatty acids composition and glutathione redox- system of male Wistar...To evaluate the effects of high fat food consisted of tallow (19% of total diets) and cholesterol (2%) on modification of erythrocyte phospholipids, fatty acids composition and glutathione redox- system of male Wistar rats with alimentary dysli- pidemia. The results demonstrated that after 30 and 180 days of high-fat feed erythrocyte phos- phatidylinositol and phosphatidylcholine levels were reduced, phosphatidylserine were in-creased. Only on the 90 days of the experiment phosphatidylinositol level increased. In all grow- ups the erythrocyte 18:0 saturated fatty acids and 20:4n6, 22:4n6 polyunsaturated fatty acids (PUFA) were increased. Deficit of n3 PUFA- 20:5n3 and 22:6n3 after 90 and 180 days high fat feed promoted compensatory synthesis from 18:1n9 on 20:3n9. Erythrocyte maleic dialde-hyde increased, glutathione level decreased in all groups of rats after fed with high-fat feed. Glutathione reductase and glutathione peroxi-dase activity decreased in erythrocytes after 30 and 180 days of high-fat feed. In conclusion: high-fat diet during 30-90 days started adaptive answer in lipids of membrane and glutathione redox-system. Important mechanism of adapta-tion of a cellular membrane to high-fat diet is increase major, structuring a membrane phos-phatidylethanolamine and minor, most meta-bolic significant fractions phospholipids (phos- phatidylinositol), keeps homeostasis of 18:2n6 and 22:6n3, 20:3n9 compensatory synthesis, decrease in activity of processes lipid peroxi-dation, activation of enzymes of redox-system glutathione. But prolonging the high-fat feeding (180 days and more) formed failure compensa-tory processes (dysadaptation). It is a risk factor of developmening atherosclerosis, diabetes, steatogepatitis and other diseases.展开更多
Although Yarrowia lipolytica is an important host strain, there have so far been few studies on the production of glutathione by the strain. We therefore performed a study to obtain an improved strain of Y. lipolytica...Although Yarrowia lipolytica is an important host strain, there have so far been few studies on the production of glutathione by the strain. We therefore performed a study to obtain an improved strain of Y. lipolytica ATCC20688, which could produce a high yield of glutathione. First, the capability of glutathione production in the ATCC20688 strain was estimated. In comparison with other yeasts, the yield of this strain was higher than those in Pichia strains. Furthermore, this strain could produce glutathione by assimilating sodium oleate. We next performed mutation and gene cloning to improve the yield. After the yield of glutathione was improved in the isolated methylglyoxal-resistant mutant (MGR3), the glutathione synthetase gene was cloned into the MGR3 strain. By using this recombinant strain, we could reach the maximum yield and intracellular content of glutathione of 54 mg/L-medium and 30 mg/g-dry cell weight, respectively.展开更多
The effects of riboflavin deficiency and simultaneously nitrosodimethylamine (NDMA) given by gastric intubation on the hepatic glutathione (GSH) content were examined in rats. On different days of the experiment, hepa...The effects of riboflavin deficiency and simultaneously nitrosodimethylamine (NDMA) given by gastric intubation on the hepatic glutathione (GSH) content were examined in rats. On different days of the experiment, hepatic GSH content of the riboflavin deficient rats decreased to 55-61% of the control rats. When NDMA was given 6 mg kg by gastric intubation to riboflavin deficient rats, hepatic GSH content decreased markedly to 39-43% of the control rats. After supplying riboflavin, hepatie GSH content of the deficient rats recovered to the level of the control rats. These results suggest that alterations of rat hepatic GSH content during riboflavin deficiency may imply as one of the promoting effects of riboflavin deficiency on the carcinogenesis of nitrosamines.展开更多
The effect of reduced glutathione (GSH) on fresh and pre-proofed frozen dough rheological properties were investigated using dynamic stress rheometry and small scale extensibility with the addition of three levels (80...The effect of reduced glutathione (GSH) on fresh and pre-proofed frozen dough rheological properties were investigated using dynamic stress rheometry and small scale extensibility with the addition of three levels (80×10-6, 160× 10-6 and 240×10-6 GSH) and six storage times (0 and 1 day, 2, 4, 6 and 8 weeks). Three relaxation times (1, 13 and 26min) after loading the dough in the rheometer were used to determine storage (G’) and loss (G”) moduli. Correlations for G’ (r=0.678 and 0.622 at 0.05, and 10Hz, respectively) and G” (r=0.699, and 0.690 at 0.05, and 10Hz, respectively) were observed with the area under the extension curve at 26 min relaxation time. The addition of GSH to fresh dough reduced G’ (16.4% to 55.9%) and G” (13.7% to 52.2%). Freezing and frozen storage caused increase in G’ and G”. The addition of GSH reduced dough strength indicated by the reduction in maximum resistance to extension (Rmax) and the ratio of maximum resistance to extensibility (Rmax/E). The reduction in Rmax across all relaxation times ranged from 16.2% to 59.4%. An increase in dough extension (E) was observed with 240×10-6 GSH at all frozen storage and rest period times. Addition of GSH caused an increase of liquid phase (30.6% to 35.3%) in fresh dough and frozen dough (10.3% to 20.7%) after one day frozen storage. Negative correlations of water content in the solid phase with dough extensibility and area under the extensibility curve were found (r=-0.594 and-0.563, respectively, p<0.001). This suggests a loss of dough extensibility and strength as the water holding capacity of the dough components changes during frozen storage.展开更多
<span style="font-family:Verdana;">The effects of each of the flavonoids;genistein (G), quercetin (Q) and</span><span style="font-family:""><span style="font-family:V...<span style="font-family:Verdana;">The effects of each of the flavonoids;genistein (G), quercetin (Q) and</span><span style="font-family:""><span style="font-family:Verdana;"> kaempferol (K) at several doses on lipid peroxides (LP) and reduced glutathione (GSH) in pooled human liver microsomes (HLMs) were investigated following the oxidative damage for 4, 6, 18 and 24 hr. HLMs (1 mg/ml) were exposed to each of the above flavonoids at 0, 5, 10, 15, 20 or 25 μM and incubated for the respective times as previously stated. Our hypothesis was that HLMs exposed to the flavonoids for the respective exposure times can decrease LP and increase GSH in HLMs to better cope with the oxidative stress. </span><span style="font-family:Verdana;">The results of our studies indicate that each of the flavonoids significantly (p < 0.01) decreased LP compared to their respective controls. The highest decrease in LP was observed for K followed by Q and G. Significant increases (p < 0.01) in GSH were observed for the flavonoid doses tested with the highest</span><span style="font-family:Verdana;"> levels observed for Q for the 24-hr. incubation. The findings suggest that the flavonoids modulate oxidative stress in HLMs by decreasing LP and such decreases in LPs may be due to the increasing and or the replenished levels of GSH in the said cells to better cope with the oxidative stress.</span></span>展开更多
基金Supported by the National Natural Science Foundation of China(No.20903045)the Specialized Research Fund for the Doctoral Program of Higher Education of China(No.20070183046)the Specialized Fund for the Basic Research of Jilin University,China(No.200810018)
文摘Glutathione transferases(GSTs) play an important role in the detoxification of xenobiotic/endobiotic toxic compounds. The α-, π-, and/l-classes of cytosolic GSTs have been studied extensively, while Gtt2 from Saccharo- myces cerevisiae, a novel atypical GST, is still poorly understood. In the present study, we investigated the gluta- thione(GSH) activation mechanism of Gtt2 using the density functional theory(DFT) with the hybrid functional B3LYP. The computational results show that a water molecule could assist a proton transfer between the GSH thiol and the N atom of His133. The energy barrier of proton transfer is 46.0 kJ/mol. The GSH activation mechanism and the characteristics of active site are different from those of classic cytosolic GSTs.
基金Supported by Scientific Research Project of Liaoning Educational Department(20060154)Initial Funds for Doctors in Dalian Nationalities University(20066206)~~
文摘[Objective] The aim of this study was to screen Saccharomyces for glutathione over-production. [Method] Ethionine-resistant mutants were obtained through UV mutagenesis and rational screening. [Result] A high GSH-producing strain HSJB1 was isolated from soil, and the biomass for this strain by flask shaking fermentation was 3.87 g/L while the GSH yield was 91.87 mg/L. According to the morphological, physiological and biochemical characteristics of cells, this strain was primarily identified as Saccharomyces cerevisiae. An ethionine-resistant mutant YBS77 was obtained through UV mutagenesis of the original strain HSJB1, and the biomass for this strain by flask shaking fermentation was 7.60 g dry cell weight/L while the GSH yield was 211.96 mg/L. [Conclusion] The biomass of the mutant obtained by breeding is increased by 96.38% than that of the original strain, and the GSH yield of the mutant obtained by breeding is increased by 130.72% than that from the original strain, which indicates that the breeding method is feasible.
文摘To evaluate the effects of high fat food consisted of tallow (19% of total diets) and cholesterol (2%) on modification of erythrocyte phospholipids, fatty acids composition and glutathione redox- system of male Wistar rats with alimentary dysli- pidemia. The results demonstrated that after 30 and 180 days of high-fat feed erythrocyte phos- phatidylinositol and phosphatidylcholine levels were reduced, phosphatidylserine were in-creased. Only on the 90 days of the experiment phosphatidylinositol level increased. In all grow- ups the erythrocyte 18:0 saturated fatty acids and 20:4n6, 22:4n6 polyunsaturated fatty acids (PUFA) were increased. Deficit of n3 PUFA- 20:5n3 and 22:6n3 after 90 and 180 days high fat feed promoted compensatory synthesis from 18:1n9 on 20:3n9. Erythrocyte maleic dialde-hyde increased, glutathione level decreased in all groups of rats after fed with high-fat feed. Glutathione reductase and glutathione peroxi-dase activity decreased in erythrocytes after 30 and 180 days of high-fat feed. In conclusion: high-fat diet during 30-90 days started adaptive answer in lipids of membrane and glutathione redox-system. Important mechanism of adapta-tion of a cellular membrane to high-fat diet is increase major, structuring a membrane phos-phatidylethanolamine and minor, most meta-bolic significant fractions phospholipids (phos- phatidylinositol), keeps homeostasis of 18:2n6 and 22:6n3, 20:3n9 compensatory synthesis, decrease in activity of processes lipid peroxi-dation, activation of enzymes of redox-system glutathione. But prolonging the high-fat feeding (180 days and more) formed failure compensa-tory processes (dysadaptation). It is a risk factor of developmening atherosclerosis, diabetes, steatogepatitis and other diseases.
文摘Although Yarrowia lipolytica is an important host strain, there have so far been few studies on the production of glutathione by the strain. We therefore performed a study to obtain an improved strain of Y. lipolytica ATCC20688, which could produce a high yield of glutathione. First, the capability of glutathione production in the ATCC20688 strain was estimated. In comparison with other yeasts, the yield of this strain was higher than those in Pichia strains. Furthermore, this strain could produce glutathione by assimilating sodium oleate. We next performed mutation and gene cloning to improve the yield. After the yield of glutathione was improved in the isolated methylglyoxal-resistant mutant (MGR3), the glutathione synthetase gene was cloned into the MGR3 strain. By using this recombinant strain, we could reach the maximum yield and intracellular content of glutathione of 54 mg/L-medium and 30 mg/g-dry cell weight, respectively.
文摘The effects of riboflavin deficiency and simultaneously nitrosodimethylamine (NDMA) given by gastric intubation on the hepatic glutathione (GSH) content were examined in rats. On different days of the experiment, hepatic GSH content of the riboflavin deficient rats decreased to 55-61% of the control rats. When NDMA was given 6 mg kg by gastric intubation to riboflavin deficient rats, hepatic GSH content decreased markedly to 39-43% of the control rats. After supplying riboflavin, hepatie GSH content of the deficient rats recovered to the level of the control rats. These results suggest that alterations of rat hepatic GSH content during riboflavin deficiency may imply as one of the promoting effects of riboflavin deficiency on the carcinogenesis of nitrosamines.
基金Financial Support of Oklahorna Wheat Foundation, Oklahome Wheat Commission and Oklahoma Experiment Station of Oklahoma State University.
文摘The effect of reduced glutathione (GSH) on fresh and pre-proofed frozen dough rheological properties were investigated using dynamic stress rheometry and small scale extensibility with the addition of three levels (80×10-6, 160× 10-6 and 240×10-6 GSH) and six storage times (0 and 1 day, 2, 4, 6 and 8 weeks). Three relaxation times (1, 13 and 26min) after loading the dough in the rheometer were used to determine storage (G’) and loss (G”) moduli. Correlations for G’ (r=0.678 and 0.622 at 0.05, and 10Hz, respectively) and G” (r=0.699, and 0.690 at 0.05, and 10Hz, respectively) were observed with the area under the extension curve at 26 min relaxation time. The addition of GSH to fresh dough reduced G’ (16.4% to 55.9%) and G” (13.7% to 52.2%). Freezing and frozen storage caused increase in G’ and G”. The addition of GSH reduced dough strength indicated by the reduction in maximum resistance to extension (Rmax) and the ratio of maximum resistance to extensibility (Rmax/E). The reduction in Rmax across all relaxation times ranged from 16.2% to 59.4%. An increase in dough extension (E) was observed with 240×10-6 GSH at all frozen storage and rest period times. Addition of GSH caused an increase of liquid phase (30.6% to 35.3%) in fresh dough and frozen dough (10.3% to 20.7%) after one day frozen storage. Negative correlations of water content in the solid phase with dough extensibility and area under the extensibility curve were found (r=-0.594 and-0.563, respectively, p<0.001). This suggests a loss of dough extensibility and strength as the water holding capacity of the dough components changes during frozen storage.
文摘<span style="font-family:Verdana;">The effects of each of the flavonoids;genistein (G), quercetin (Q) and</span><span style="font-family:""><span style="font-family:Verdana;"> kaempferol (K) at several doses on lipid peroxides (LP) and reduced glutathione (GSH) in pooled human liver microsomes (HLMs) were investigated following the oxidative damage for 4, 6, 18 and 24 hr. HLMs (1 mg/ml) were exposed to each of the above flavonoids at 0, 5, 10, 15, 20 or 25 μM and incubated for the respective times as previously stated. Our hypothesis was that HLMs exposed to the flavonoids for the respective exposure times can decrease LP and increase GSH in HLMs to better cope with the oxidative stress. </span><span style="font-family:Verdana;">The results of our studies indicate that each of the flavonoids significantly (p < 0.01) decreased LP compared to their respective controls. The highest decrease in LP was observed for K followed by Q and G. Significant increases (p < 0.01) in GSH were observed for the flavonoid doses tested with the highest</span><span style="font-family:Verdana;"> levels observed for Q for the 24-hr. incubation. The findings suggest that the flavonoids modulate oxidative stress in HLMs by decreasing LP and such decreases in LPs may be due to the increasing and or the replenished levels of GSH in the said cells to better cope with the oxidative stress.</span></span>