1.Classification of Peroxiredoxin Subfamilies Using Regular Expressions.
Jae Kyung CHON ; Jongkeun CHOI ; Sang Soo KIM ; Whanchul SHIN
Genomics & Informatics 2005;3(2):55-60
Peroxiredoxins (Prx's) are a superfamily of peroxidases that are ubiquitous in all super-kingdoms. Previous biochemical and structural studies have suggested that Prx's could be divided into five subfamilies (1-Cys, Typical 2-Cys, Atypical 2-Cys C-, L- and R- types). In this work, we have developed a set of regular expression patterns describing subfamily-specific spatial constraints of the key catalytic residues. Using these patterns, 1,016 Prx's available in public databases were classified into the five subfamilies. Our method performed well for most of the types except for Atypical 2 Cys R type.
Classification*
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Peroxidases
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Peroxiredoxins*
2.Study on effect of drought stress on physiological adaptation of seedlings of Atractylodes lancea.
Jie ZHOU ; Lu-Qi HUANG ; Lan-Ping GUO ; Yong-Qing ZHANG
China Journal of Chinese Materia Medica 2008;33(19):2163-2166
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
3.Construction of eukaryotic expression plasmid of human PRX3 and its expression in HEK-293FT cells.
Yan, FENG ; Zhao, LIU ; Huiqing, CAO ; Xianmin, MENG ; Zhiling, QU ; Mi, XIONG ; Zhongduan, DENG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2004;24(4):311-3, 321
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
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Cloning, Molecular
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Embryo
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Eukaryotic Cells/*metabolism
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Gene Expression
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Genetic Vectors
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Kidney/cytology
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Kidney/*metabolism
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Peroxidases/*biosynthesis
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Peroxidases/genetics
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Plasmids/*genetics
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Transfection
4.Optimization of manganese peroxidase production from Schizophyllum sp. F17 in solid-state fermentation of agro-industrial residues.
Yue ZHOU ; Bing YANG ; Yang YANG ; Rong JIA
Chinese Journal of Biotechnology 2014;30(3):524-528
Manganese peroxidase (MnP), a crucial enzyme in lignin degradation, has wide potential applications in environmental protection. However, large-scale industrial application of this enzyme is limited due to several factors primarily related to cost and availability. Special attention has been paid to the production of MnP from inexpensive sources, such as lignocellulosic residues, using solid-state fermentation (SSF) systems. In the present study, a suitable SSF medium for the production of MnP by Schizophyllum sp. F17 from agro-industrial residues has been optimized. The mixed solid medium, comprising pine sawdust, rice straw, and soybean powder at a ratio of 0.52:0.15:0.33, conferred a maximum enzyme activity of 11.18 U/g on the sixth day of SSF. The results show that the use of wastes such as pine sawdust and rice straw makes the enzyme production more economical as well as helps solve environmental problems.
Culture Media
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Fermentation
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Industrial Microbiology
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methods
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Oryza
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Peroxidases
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biosynthesis
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Schizophyllum
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enzymology
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Wood
5.Impact of exogenous paraquat on enzyme exudation and biochemical changes of lignin degradation fungi.
Yunchen ZHAO ; Jianlong LI ; Yuru CHEN ; Haixia HUANG ; Zui YU
Chinese Journal of Biotechnology 2009;25(8):1144-1150
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
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drug effects
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enzymology
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Lignin
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metabolism
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Oxidative Stress
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Paraquat
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pharmacology
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Peroxidases
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metabolism
6.Heterologous Expression of Phanerochaete chrysoporium Glyoxal Oxidase and its Application for the Coupled Reaction with Manganese Peroxidase to Decolorize Malachite Green.
Yu Lim SON ; Hyoun Young KIM ; Saravanakumar THIYAGARAJAN ; Jing Jing XU ; Seung Moon PARK
Mycobiology 2012;40(4):258-262
cDNA of the glx1 gene encoding glyoxal oxidase (GLX) from Phanerochaete chrysosporium was isolated and expressed in Pichia pastoris. The recombinant GLX (rGLX) produces H2O2 over 7.0 nmol/min/mL using methyl glyoxal as a substrate. Use of rGLX as a generator of H2O2 improved the coupled reaction with recombinant manganese peroxidase resulting in decolorization of malachite green up to 150 microM within 90 min.
Alcohol Oxidoreductases
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DNA, Complementary
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Glyoxal
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Manganese
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Organometallic Compounds
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Oxidoreductases
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Peroxidase
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Peroxidases
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Phanerochaete
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Pichia
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Rosaniline Dyes
7.Induction of Defense Related Enzymes and Pathogenesis Related Proteins in Pseudomonas fluorescens-Treated Chickpea in Response to Infection by Fusarium oxysporum f. sp. ciceri.
Ratul SAIKIA ; Rakesh KUMAR ; Tanuja SINGH ; Alok K SRIVASTAVA ; Dilip K ARORA ; Min Woong LEE
Mycobiology 2004;32(1):47-53
Pseudomonas fluorescens 1-94 induced systemic resistance in chickpea against Fusarium wilt of chickpea caused by Fusarium oxysporum f. sp. ciceri by the synthesis and accumulation of phenolic compounds, phenylalanine ammonia lyase(PAL) and pathogenesis related(PR) proteins(chitinase, beta-1,3-glucanase and peroxidase). Time-course accumulation of these enzymes in chickpea plants inoculated with P. fluorescens was significantly(LSD, P=0.05) higher than control. Maximum activities of PR-proteins were recorded at 3 days after inoculation in all induced plants; thereafter, the activity decreased progressively. Five PR peroxidases detected in induced chickpea plants. Molecular mass of these purified peroxidases was 20, 29, 43, 66 and 97 kDa. Purified peroxidases showed antifungal activity against plant pathogenic fungi.
Ammonia
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Cicer*
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Fungi
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Fusarium*
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Peroxidases
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Phenol
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Phenylalanine
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Plants
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Pseudomonas fluorescens
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Pseudomonas*
8.Purification and characterization of a bromoperoxidase from Gracilaria lemaneiformis.
Haiyan LI ; Yan JIN ; Wei ZHANG ; Xingju YU ; Jinyou ZHANG ; Peichun WU
Chinese Journal of Biotechnology 2008;24(4):622-626
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
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methods
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Enzyme Stability
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Gracilaria
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enzymology
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Hydrogen-Ion Concentration
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Kinetics
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Peroxidases
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isolation & purification
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metabolism
9.Light quality regulation of growth and endogenous IAA metabolism of Ganoderma lucidum mycelium.
Xi-Ling MEI ; Zhou ZHAO ; Xiang-Dong CHEN ; Jin LAN
China Journal of Chinese Materia Medica 2013;38(12):1887-1892
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
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analysis
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Indoleacetic Acids
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metabolism
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Light
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Peroxidases
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metabolism
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Reishi
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growth & development
;
metabolism
10.Expression of Glutathione Peroxidases and Its Effect on Clinical Prognosis in Glioma Patients.
Xiao-Mei REN ; Li ZHANG ; Bao XIN ; Wen-Wen QIAN ; Zeng-Run XIA ; Meng QI ; Xiao-Ping DU ; Chuan-Dao SHI ; Qi-Ling LIU ; Rong-Qiang ZHANG
Acta Academiae Medicinae Sinicae 2022;44(2):276-285
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
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Glioblastoma
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Glioma/diagnosis*
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Glutathione Peroxidase/metabolism*
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Humans
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Peroxidases
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Prognosis
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Proportional Hazards Models