OBJECTIVETo study the effect of drought stress on the changes of physiological adaptation of Atractylodes lancea seedlings.

METHODInvestigation was carried out on content changes of MDA, soluble protein, and activities of SOD, POD, CAT, APX in A. lancea seedlings under polyethylene glycol (PEG-6000)-simulated drought stress.

RESULTIn A. lancea seedlings treated with 15% and 25% PEG, the content of MDA increased significantly with the stress time, and increased more significantly at a higher concentration of PEG. The content of soluble protein increased significantly after treatment on the day one and day three; activities of SOD, POD, CAT and APX increased at first and decreased later, increasing rates rose at high concentration of PEG moreover, activities of POD, APX reached the maximum after three days, and the time of maximum activities changed with concentration of PEG.

CONCLUSIONA. lancea seedlings adapted to drought stress by increasing the content of soluble protein to decrease water potential, and by improving activities of protective enzymes to enhance anti-oxidative ability under drought stress.

Adaptation, Physiological ; physiology ; Ascorbate Peroxidases ; Atractylodes ; enzymology ; metabolism ; physiology ; Catalase ; metabolism ; Droughts ; Gene Expression Regulation, Plant ; physiology ; Peroxidase ; metabolism ; Peroxidases ; metabolism ; Seedlings ; enzymology ; metabolism ; physiology ; Superoxide Dismutase ; metabolism

To study the effect and mechanism of the light quality acting on Ganoderma lucidum, and provide a theoretical basis for G. lucidum mycelium cultivation, we focused on growth and endogenous IAA metabolism of G. lucidum mycelium under different light-emitting diode (LED) condition. The growth index, endogenous levels of IAA and Enzymes related to IAA metabolism and Polysaccharides content were investigated in different growth periods. Results showed that blue light irradiation was the best from the viewpoint of steady growth and polysaccharides accumulation, red light irradiation improved endogenous IAA level and promoted growth of mycelium in early stage of cultivation, green light irradiation decreased growth rate and fresh weight of mycelium, but increased drying rate. Enzymes related to IAA metabolism also significantly influenced by light quality. The activity of indole acetic acid oxidase (IAAO), peroxidase (POD) and tryptophan synthetase with blue light irradiation were showed high level in early time, but decreased later, and the IAA content was consistently at lower level than that in other treatments, while mycelium irradiated with yellow light showed the highest activity of both IAAO and tryptophan synthetase, and medium level of IAA content. In conclusion, the light quality affects growth and regulation of the level of endogenous IAA of G. lucidum mycelium.
Fungal Polysaccharides ; analysis ; Indoleacetic Acids ; metabolism ; Light ; Peroxidases ; metabolism ; Reishi ; growth & development ; metabolism

To construct the eukaryotic expression plasmid of human PRX3 and measure its expression in the HEK-293FT cells, the full-length coding region of human PRX3 was cloned by PCR and inserted into the eukaryotic expression vector pcDNA4-Xpress (A). HEK-293FT cells were transiently transfected with the recombinant plasmid. Western blot and immuofluorescence were used to detect the expression of the fusion protein. In the experiment, restriction analysis identified the construction of the recombinant plasmid and the inserted sequence was identical with that published on GenBank. Western blot and immunofluorescence confirmed the expression of the recombinant protein in transfected HEK-293FT cells. It was concluded that the eukaryotic expression plasmid of human PRX3 was constructed successfully and the recombinant could be expressed efficiently in HEK-293FT cells, which provides a sound basis for the further study on human PRX3.
Cell Line, Transformed ; Cloning, Molecular ; Embryo ; Eukaryotic Cells/*metabolism ; Gene Expression ; Genetic Vectors ; Kidney/cytology ; Kidney/*metabolism ; Peroxidases/*biosynthesis ; Peroxidases/genetics ; Plasmids/*genetics ; Transfection

To study the effect of exogenous oxygen, we added water solution of paraquat to 7 d cultures of Coriolus versicolor for the next 148 h. Enzyme exudation and biochemical process were investigated on the addition of paraquat. We found that compared with the control (without paraquat), the addition of 30 micromol/L paraquat stimulated the activity of manganese dependent peroxidase (MnP), lignin peroxidase (LiP), and laccases (Lac) 7, 2.5 and 1.3 times, respectively. Also, addition of paraquat enhanced activity of superoxide dismutase (SOD) and catalase (CAT) in the first 48 h. Impact of paraquat on ligninolytic enzymes was significant than that on antioxidant enzyme. Addition of paraquat enhanced phenolic compounds and formaldehyde of cultures too. And concentration of malondialdehyde was increased in the first 24 h. The results showed that addition of paraquat promoted oxidative stress, but the antioxidant systems of the fungal strain are sufficient to prevent mycelia from oxidative stress. As exogenous oxygen, paraquat might be a useful substrate in degradation of lignocellulose.
Fungi ; drug effects ; enzymology ; Lignin ; metabolism ; Oxidative Stress ; Paraquat ; pharmacology ; Peroxidases ; metabolism

A bromoperoxidase from Gracilaria lemaneiformis was purified to homogeneity using a multi-step process of ammonium sulfate precipitation (AS), dialysis, and DEAE-cellulose 52 anion exchange chromatography. The bromoperoxidase activity was unstable or undetectable in crude extract solution. However, it became stable with electrophoretic purity after this multiple purification process. The anion exchange chromatography purification was a critical step in the purification process, which effectively eliminated the phycobiliprotein and smucilaginous polysaccharides. The purified bromoperoxidase was a monomeric enzyme with the relative molecular masses of 66 kD as determined by denaturing and native gradient gel electrophoresis. The optimal pH for bromoination was 6.0 and bromoperoxidase activity was stable as stored at a broad pH range of 3.0-9.0. Of a range of compounds tested, only vanadium enhanced bromoperoxidase activity. Kinetic studies for the bromination of monochlorodimedone (MCD) showed that the Km values of Br- and H2O2 are 53.5 micromol/L, 38 micromol/L respectively.
Chromatography, Ion Exchange ; methods ; Enzyme Stability ; Gracilaria ; enzymology ; Hydrogen-Ion Concentration ; Kinetics ; Peroxidases ; isolation & purification ; metabolism

Objective To investigate the relationship between the expression of glutathione peroxidase(GPX)genes and the clinical prognosis in glioma patients,and to construct and evaluate the model for predicting the prognosis of glioma. Methods The clinical information and GPX expression of 663 patients,including 153 patients of glioblastoma(GBM)and 510 patients of low-grade glioma(LGG),were obtained from The Cancer Genome Atlas(TCGA)database.The relationship between GPX expression and patient survival was analyzed.The key GPX affecting the prognosis of glioma was screened out by single- and multi-factor Cox's proportional-hazards regression models and validated by least absolute shrinkage and selection operator(Lasso)regression.Finally,we constructed the model for predicting the prognosis of glioma with the screening results and then used concordance index and calibration curve respectively to evaluate the discrimination and calibration of model. Results Compared with those in the control group,the expression levels of GPX1,GPX3,GPX4,GPX7,and GPX8 were up-regulated in glioma patients(all P<0.001).Moreover,the expression levels of other GPX except GPX3 were higher in GBM patients than in LGG patients(all P<0.001).The Kaplan-Meier curves showed that the progression-free survival of GBM with high expression of GPX1(P=0.013)and GPX4(P=0.040),as well as the overall survival,disease-specific survival,and progression-free survival of LGG with high expression of GPX1,GPX7,and GPX8,was shortened(all P<0.001).GPX7 and GPX8 were screened out as the key factors affecting the prognosis of LGG.The results were further used to construct a nomogram model,which suggested GPX7 was the most important variable.The concordance index of the model was 0.843(95%CI=0.809-0.853),and the calibration curve showed that the predicted and actual results had good consistency. Conclusion GPX7 is an independent risk factor affecting the prognosis of LGG,and the nomogram model constructed with it can be used to predict the survival rate of LGG.
Brain Neoplasms ; Glioblastoma ; Glioma/diagnosis* ; Glutathione Peroxidase/metabolism* ; Humans ; Peroxidases ; Prognosis ; Proportional Hazards Models

Basidiomycete PM2, a lignin-degrading white rot fungus, produces lgnin peroxidase (Lip) and manganese peroxidase (Mnp) in nutrient nitrogen limited liquid cultures. This fungus was selected for its ability to decolorize azo group of dyes. In order to improve production of the peroxidases and rapid dye decolorizing activity by basidiomycete PM2, the addition of veratryl alcohol or Tween 80 to nutrient nitrogen limited liquid cultures were tested. It was found to have a large stimulatory effect on Mnp activities and decolorization rate of azo dyes. A maximum Mnp activities of 254.2 u/L with veratryl alcohol and 192.2 u/L with Tween 80 were achieved respectively. These values were about 3.4-fold and 2.5-fold higher than that obtained in the control cultures (without alcohol or Tween 80), whereas the levels of Lip activity detected were very low (about 12 u/L)in all the cultures. In further experiments using three kinds of azo dyes of congo red, orange G and orange IV, enzyme activities and dye decolorization were investigated in the above-mentioned cultures. The results showed that Mnp activities and decolorization were notably higher than those obtained in the control cultures in the presence of azo dyes. Cultures supplemented with Tween 80 were more adequate for dye decolorization. The rates of the decolorization with Tween 80 of congo red (95.4%), orange G (98.5%) and orange IV (54.4%) after 24 hours of dye incubation were higher than that supplemented with veratryl alcohol. According to the results, Mnp activities secreted by basidiomycete PM2 play an essential role in the process of dye decolorization. Tween 80 was the main factor affecting the decolorization. The analysis of structure of the three kinds of azo dyes indicats that the extent of decolorization is affected by the dye molecular structure. The types and quantity of the substituted groups on the aromatic ring of azo dyes have effect on the percentage of biological decolorization.
Azo Compounds ; metabolism ; Basidiomycota ; enzymology ; metabolism ; Benzyl Alcohols ; pharmacology ; Coloring Agents ; metabolism ; Oxygenases ; biosynthesis ; Peroxidases ; biosynthesis ; Polysorbates ; pharmacology

Wheat peroxidases 1 (WP1) is the major cationic peroxidase of wheat (Triticum aestivum) grain, which is involved in the development of seeds and an important factor to affect the final processing quality of flour. We constructed a prokaryotic expression vector pET28a-WP1, and transformed it into E. coli host strain T7 Expression. His-tag fused WP1 existed as inclusion body, and the recombinant protein was purified by Ni-NTA resin affinity chromatography under denatured condition. The purity of target protein reached 98%. The recombinant WP1 was refolded by gradient urea dialysis, then used as antigen to immune rabbit to prepare polyclonal antibody. The result of ELISA showed that the titer of rabbit anti-WP1 antiserum was higher than 1:625 000. The result of Western blotting demonstrated that the prepared WP1 polyclonal antibody could be used to detect WP1 with high specificity.
Animals ; Antibodies ; immunology ; metabolism ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Peroxidases ; biosynthesis ; genetics ; Rabbits ; Recombinant Proteins ; biosynthesis ; genetics ; immunology

The dynamic changes of germination percentage, germination potential, thousand-seed weight, antioxidase activity in Desmodium styracifolium seeds with different storage time were tested, and electrical conductivity, contents of soluble sugar, soluble protein, starch in seed leach liquor were also determined in order to reveal the mechanism of seed deterioration. The results as the following. (1) The germination percentage, germination potential and thousand-seed weight of D. styracifolium seeds declined, while the seed coat color darkened with the extension of storage time. (2) The activities of superoxide dismutase (SOD) and peroxidase (POD) decreased with the prolongation of storage period. The SOD activity declined fastest in 1,095-1,185 d of storage, while the POD activity declined significantly in 365-395 d of storage. (3) The electrical conductivity and the contents of soluble sugar, starch in seed leach liquor increased, while the content of soluble protein declined with the extension of storage time. (4) Correlation analysis indicated that the germination percentage, germination potential and thousand-seed weight of D. styracifolium seeds have a significantly positive correlation with SOD and POD activity, while have a significantly negative correlation with the electrical conductivity, contents of soluble sugar and starch. It can be concluded that during the storage of D. styracifolium seeds, physiological and biochemical changes including decrease in antioxidase activity, rise in electrical conductivity, degradation effluent of soluble sugar and starch, degradation of soluble protein were the main factors leading to the seed deterioration.
Color ; Fabaceae ; chemistry ; enzymology ; growth & development ; metabolism ; Germination ; Peroxidases ; metabolism ; Plant Proteins ; metabolism ; Seeds ; chemistry ; enzymology ; growth & development ; metabolism ; Starch ; metabolism ; Superoxide Dismutase ; metabolism ; Time Factors

Lignin peroxidase (LiP) hosted in Brij 30/cyclohexane/water nonionic reversed micelle could express its catalytic activity, but in Triton X-100/n-pentanol/cyclohexane/water nonionic reversed micelle LiP didn't show any catalytic activity. Some key factors that affected the catalytic activity of LiP in Brij 30 reversed micelle were studied at 20 degrees C. The optimum conditions were:omega0 = 8.5, pH = 2.2, [Brij30] = 600 mmol/L; under these conditions the half time of LiP was ca. 50 hours. As compared with the properties of LiP in aqueous solution, the activity of LiP hosted in Brij 30 reversed micelle dropped, but its stability improved greatly. To reveal the role of normal alcohol, which was a necessary component for forming Triton X-100 reversed micelles, the effect of n-pentanol on the catalytic activity of LiP in Brij 30 reversed micelle was investigated. Results indicated that high concentration of the alcohol deactivated LiP. So it was deduced that the phenomenon that LiP hosted in the Triton X-100 reversed micelles could not express its activity was mainly due to the alcohol co-surfactant.
Catalysis ; Cyclohexanes ; chemistry ; Enzyme Activation ; drug effects ; Micelles ; Octoxynol ; chemistry ; Pentanols ; chemistry ; Peroxidases ; metabolism ; Surface-Active Agents ; chemistry