• Journal of Microbiology and Biotechnology
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• v.14 no.1
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• pp.74-80
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• 2004

Aspergillus fumigatus is one of the most common fungi in the human environment, both in-doors and out-doors. It is the main causative agent of invasive aspergillosis, a life-threatening mycosis among immunocompromised patients. The genome has been sequenced by an international consortium, including the Wellcome Trust Sanger Institute (U.K.) and The Institute for Genomic Research (TIGR, U.S.A.), and a ten times whole genome shotgun sequence assembly has been made publicly available. In this study, we identified tricarboxylic acid (TCA) cycle enzymes of A. fumigatus by comparative analysis with four other fungal species. The open reading frames showed high amino acid sequence similarity with the other fungal citric acid enzymes and well-conserved functional domains. All genes present in Saccharomyces cerevisiae, Schizosaccharomyces pombe, Candida albicans, and Neurospora crassa were also found in A. fumigatus. In addition, we identified four A. fumigatus genes coding for enzymes in the glyoxylate shunt, which may be required for fungal virulence. The architecture of multi-gene encoded enzymes, such as isocitrate dehydrogenase, 2-ketoglutarate, succinyl-CoA synthetase, and succinate dehydrogenase was well conserved in A. fumigatus. Furthermore, our results show that genes of A. fumigatus can be detected reliably using GlimmerM.

• Journal of Ginseng Research
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• v.7 no.1
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• pp.68-73
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• 1983

E. coli (K-12 W1485) was grown in M9 minimal medium containing ginseng saponin (10-3%-2%) and found that the cells grew most rapidly in the presence of 10-1% saponin. The cells, harvested at the early exponential phase, were transferred to the minimal medium containing 10-1% saponin plus 14C-labelled saponin (0.03 ${\mu}$Ci) and the incubation was continued at 37$^{\circ}C$ for 20 minutes and the cells were fractionated into the outermembrane, innermembrane and cytosol fraction. Radioactivity data showed that the most radioactivity was detected in the innermembrane. The activity of succinate dehydrogenase of the cells grown in the above saponin medium was significantly higher than that of the cells grown in ordinary minimal medium. No significant difference of the glucose 6-phosphate dehydrogenase activity was observed between the two groups. It was also found that the saponin stimulated glucose uptake and biosynthisis of lipids and proteins of the cells. Incorporation of 14C-leucine into the protein fraction of the cell was also accelerated.

• Korean Journal of Acupuncture
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• v.29 no.2
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• pp.260-270
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• 2012

Objectives : Hippocampus, a region of temporal lobe, plays an important role in the pathogenic mechanisms of brain diseases such as Alzheimer's disease, depression and temporal lobe epilepsy. This research is designed to investigate hippocampal changes after acupuncture stimulation at Shinmun(HT7) using 2-dimensional gel electrophoresis(2-DE). Methods : On postnatal-day 15, rat pups were randomly devided into Normal(NOR) or HT7 group. All of Pups kept with their mothers for 7 days, but pups in HT7 group received acupuncture stimulation at HT7 daily. On postnatal-day 21, hippocampus of each rat pup was dissceted 30 minutes after last acupuncture stimulation and the protein expressions were investigated using 2-DE. Results : After acupuncture stimulation at HT7, expression of 20 proteins were significantly increased. Succinate semialdehyde dehydrogenase, glyceraldehyde-3-phosphate dehydrogenase-like, transketolase, aconitate hydratase and phosphoglucomutase-1 were related to glucose methabolism. Eukaryotic initiation factor(eIF) 4A-II, eIF 4A-III, mitochondrial Tu translation elongation factor and chain A of crystal structure of the 70-Kda heat shock cognate protein involve in the protein synthesis in ribosome. Tubulin ${\beta}$-4 chain, tubulin T ${\beta}$-15 and tubulin ${\alpha}$-1B chain comprise cytoskeleton. Glutathione S-transferase(GST) ${\omega}$-1, GST P and GST Yb-3 can reduce oxidative stress. ${\beta}$-soluble N-ethylmaleimide-sensitive fusion protein attachment protein is required for vesicular transport between the endoplasmic reticulum and the Golgi apparatus, glycerol-3-phosphate dehydrogenase plays a major role in lipid biosynthesis, creatine kinase U-type catalyses the conversion of creatine and consumes adenosine triphosphate to create phosphocreatine and adenosine diphosphate. Platelet-activating factor acetylhydrolase IB subunit alpha and voltage depedent anion-selective channel protein 2 were also increased. Conclusions : The results suggest that acupuncture stimulation at HT7 may enhance glucose and lipid metabolism, protein synthesis, cytoskeletal substance and anti-oxidative stress in hippocampus.

• Korean Journal of Microbiology
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• v.24 no.2
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• pp.133-140
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• 1986

Extracts of CO-autotrophically grown cells of Acinetobacter sp. 1 were shown to use thionin, methylene blue, or 2,6-dichlorophenol-indophenol, but not NAD, NADP, FAD, or FMN, as electron acceptors for the oxidation of CO under strictly anaerobic conditions. The CO dehydrogenase (CO-DH) in the thes bacterium was found to be an inducible enzyme. The enzyme activity was determined by an assay based on the CO-dependent reduction of thionin. Maximal reaction rates were found at pH 7.5 and $60^{\circ}C$, and the Arrhenius plot revealed an activation energy of 6.1 kcal/mol(25.5kJ/mol). THe $K_m$ m/ for CO was $154{\mu}M$. Known metalchelating agents tested had no effects on the CO-DH activity. No divalent cations tested affect the enzyme activity significantly escept $Cu^{2+}$ which suppressed the activity completely. The enzyme was inhibited by glucose and succinate. The same extracts catalyzed oxidation of hydrogen gas and formate with thionin as electron acceptor. The CO-DH of Acinetobacter sp. 1 was to have no immunological relationship with that of Pseudomonas carboxydohydrogena.

• Applied Microscopy
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• v.26 no.3
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• pp.253-266
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• 1996

To investigate the effects of mercuric chloride ($HgCl_2$) on the differentiation of the cerebral neuron of chick embryo 9 days, the ultrastructural changes in nerve cells injected with a various doses of mercuric chloride were observed with transmission electron microscope. The enzyme activity of the some dehydrogenases, cerebral proteins and adenosine triphosphate (ATP) were also analyzed. The results obtained are as follows: The ultrastructural changes in 0.5 and 1.0mg-injected groups were undetectable, but in 2.0mg-injected group, the nuclear envelops were very irregular and mitochondria, were swelled and destroyed partly. The number of polypeptide bands separated by SDS-PAGE in the normal group were 37 bands. According to the in creased dose of mercuric chloride, contends of the bands were increased in 7 bands. The activities of dehydrogenases were declined by increasing the dose of mercuric chloride. Lactate dehydrogenase (LDH) activity failed to 78% in 1.0mg-injected group and greatly to 68% in 2.0 mg-injected group. Malate dehydrogenase (MDH) activity failed to 81% in 2.0 mg-injected group. On the other hand, succinate dehydrogenase (SDH) activity decreased to 80% in 1.0 mg-injected group and greatly to 63% in 2.0 mg-injected group. ATP content in 1.0 mg-injected group was increased slightly and in 2.0 mg-injected group was increased greatly.

• Applied Microscopy
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• v.22 no.1
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• pp.89-102
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• 1992

This research was undertaken to determine the effects of the anticancer and immunosuppressive drug cyclophosphamide (CP) on the epididymal caput of the male rat in terms of ultrastructural alteration and protein analysis by SDS-PAGE at different groups; control group, 1 week group, 3 weeks group, 5 weeks group were treated with saline (control group) or CP at doses of 20 mg/kg/week, 1 time a week, respectively. In the cytoplasm of the principal cells on the epididymis, the mitochondrial outer and inner membranes were significantly swollen or disrupted. The lumens of rough endoplasmic reticulum (rER) were also swollen, and the number of Golgi vesicles were increased, respectively. It is suggested that treatment with CP alters the specific cell organelles in the epididymis. CP caused changes in protein concentrations in caput of epididymis after CP treatment. Total proteins of 32 to 37 species such as lactate dehydrogenase, carnitine acetyltransferase and succinate dehydrogenase were expressed in the caput fluid. Then the more CP was increased, the more concentration of proteins caused to decrease, synthesize or increase in epididymis. In contrast to the control group, in particular carnitine acetyltransferase and the other 9 proteins in the caput fluid were decreased or disappeared, respectively, whereas lactate dehydrogenase and the other 5 proteins in the caput fluid were increased or synthesized, respectively. The other proteins are not showed distinctive difference. These alterations could be direct mediated by toxic effects of the drug on the epithelium or be secondary to changes in the spermatozoa as a result of the CP treatment.

• Applied Microscopy
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• v.27 no.1
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• pp.87-100
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• 1997

To investigate the effects of mercuric chloride $(HgCl_2)$ on the differentiation of the cerebral neuron of chick embryo 10 days, the ultrastructural changes in nerve cells injected with a various doses of mercuric chloride were observed with transmission electron microscope. The enzyme activity of the some dehydrogenases, cerebral proteins and adenosine triphosphate (ATP) were also analyzed. The results obtained are as follows; The ultrastructural changes in 1.0 mg-injected group, the nuclear membranes were irregular, outer of mitochondria membrances dispressioned, their cristae were destroyed. In 2.0 mg-injected group, the nuclear envelops were destroyed and divided, were not observed organelle except of few ribosome, the RER and mitochondria. The number of polypeptide bands were separated by SDS-PAGE in the normal group were 38 bands. According to the in creased dose of mercuric chloride, contends of the bands were increased in 4 bands, but were decreased in 1 band. The activities of dehydrogenases were declined by increasing the dose of mercuric chloride. Lactate dehydrogenase (LDH) activity fatted to 61% in 2.0 mg-injected group. Malate dehydrogenase (MDH) activity fatted to 90% in 1.0 mg-injected group, greatly to 76% in 2.0 mg-injected group. Succinate dehydrogenase (SDH) activity decreased to 79% in 1.0 mg-injected group and greatly to 62% in 2.0 mg-injected group. ATP content in 1.0 mg-injected group was almost near to the normal level, but it was increased greatly in 2.0 mg-injected group.

• BMB Reports
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• v.35 no.5
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• pp.443-451
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• 2002

Malonate is a three-carbon dicarboxylic acid. It is well known as a competitive inhibitor of succinate dehydrogenase. It occurs naturally in biological systems, such as legumes and developing rat brains, which indicates that it may play an important role in symbiotic nitrogen metabolism and brain development. Recently, enzymes that are related to malonate metabolism were discovered and characterized. The genes that encode the enzymes were isolated, and the regulation of their expression was also studied. The mutant bacteria, in which the malonate-metabolizing gene was deleted, lost its primary function, symbiosis, between Rhizobium leguminosarium bv trifolii and clover. This suggests that malonate metabolism is essential in symbiotic nitrogen metabolism, at least in clover nodules. In addition to these, the genes matB and matC have been successfully used for generation of the industrial strain of Streptomyces for the production of antibiotics.

• Biomolecules & Therapeutics
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• v.16 no.1
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• pp.1-8
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• 2008

Mitochondria are important sensor of apoptosis. $H_2O_2-induced$ cell death rate was enhanced by serum deprivation. In this study, we investigated whether serum deprivation using 0.5 or 3 % FBS induces apoptotic cell death through mitochondrial enzyme activation as compared to 10 % FBS. Apoptotic cell death was observed by chromosome condensation and the increase of sub-G0/G1 population. Serum deprivation reduced cell growth rate, which was confirmed by the decrease of S-phase population in cell cycle. Serum deprivation significantly increased caspase-9 activity and cytochrome c release from mitochondria into cytosol. Serum deprivation-induced mitochondrial changes were also indicated by the increase of ROS production and the activation of mitochondrial enzyme, succinate dehydrogenase. Mitochondrial enzyme activity increased by serum deprivation was reduced by the treatment with rotenone, mitochondrial electron transport inhibitor. In conclusion, serum deprivation induced mitochondrial apoptotic cell death through the elevation of mitochondrial changes such as ROS production, cytochrome c release and caspase-9 activation. It suggests that drug sensitivity could be enhanced by the increase of mitochondrial enzyme activity in serum-deprived condition.

• Journal of Microbiology and Biotechnology
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• v.15 no.2
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• pp.281-286
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• 2005

We created a new graphite-Cu(II) electrode and found that the electrode could catalyze FADH$_2$ oxidation and FAD reduction coupled to electricity production and consumption, respectively. In a fuel cell with graphite-Cu(II) anode and graphite-Fe(III) cathode, the electricity was produced by coupling to the spontaneous oxidation of FADH$_2$ Fumarate and xylose were not produced from the enzymatic oxidation of succinate and xylitol without FAD, respectively, but produced with FAD. The production of fumarate and xylose in the reactor with FAD electrochemically regenerated was maximally 2- 5 times higher than that in the reactor with FAD. By using this new electrode with catalytic function, a bioelectrocatalysts can be engineered; namely, oxidoreductase (e.g., lactate dehydrogenase) and FAD can function for biotransformation without an electron mediator and second oxidoreductase for cofactors recycling.