• Journal of Environmental Science International
• /
• v.17 no.11
• /
• pp.1221-1226
• /
• 2008

A variety of fungal species are known to degrade cyanide through the action of cyanide hydratase, a specialized nitrilases which hydrolyze cyanide to formamide. This work is a report on two unknown and un-characterized members from Neurospora crassa and Aspergillus nidulans. Recombinant forms of three cyanide hydratases (CHT) originated from N. crassa, Gibberella zeae, and A. nidulans were prepared after their genes were cloned with N-terminal hexahistidine purification tags, expressed in E. coli and purified using immobilized metal affinity chromatography. These enzymes were compared according to their pH activity profiles, and kinetic parameters. Although all three were similar, the N. crassa CHT has the widest pH range of activity above 50% and highest turnover rate ($6.6{\times}10^8min^{-1}$) among them. The CHT of A. nidulans has the highest Km value of the three nitrilases evaluated in here. Expression of CHT in both N. crassa and A. nidulans were induced by the presence of KCN, regardless of any presence of nitrogen sources. These data can be used to determine optimal procedures for the enzyme uses in the remediation of cyanide-containing wastes.

• Journal of Plant Biology
• /
• v.33 no.2
• /
• pp.135-140
• /
• 1990

The membrane potential in the subepidermal cells of Lemna gibba G3 fronds was measured in the dark with glass capillary microelectrodes. At pH 7, the membrane potential, approximately-215 mV, could be depolarized to -82∼-88 mV by 0.1 mM dicyclohexylcarbodiimide (DCCD) or by KCN at 0.3 mM or higher concentrations. When the pH of the medium was altered the potential showed reversible changes, while it revealed no response to the external pH changes when energy transduction across the membrane was being blocked by 0.1 mM DCCD. The results support an assumption that the active component of the membrane potential of Lemna subepidermal cells is generated by electrogenic H+ -pump. By the addition of 0.10∼5.00 mM salicylic acid(SA) to the bathing medium the membrane potential was depolarized to a great extent, and the removal of SA from the medium repolarized the potential showing almost complete recovery, 92.3∼97.6% to the initial levels. Although the potential was greatly depolarized by 5.0% or higher concentrations of dimethylsulfoxide (DMSO), the recovery rate by DMSO removal was decreased as the pretreatment concentration had increased. Twenty percent DMSO pretreatment limited the recovery at only 47.1%. The presence of SA in the bathing medium could reversibly increase the permeability of the plasmalemma. DMSO at its concentration of 5.0% or higher increased the permeability of the membrane by irrevesibly impairing the membrane component involved in the membrane permeability.

• Biomedical Science Letters
• /
• v.20 no.2
• /
• pp.96-102
• /
• 2014

Staphylococcal enterotoxin B (SEB) is bacterial toxin that induces the activation of immune cells. Because the inhibition of pro-inflammatory effect of SEB can resolve the inflammation, I determined the influence of functional or structural change of SEB on immune cells. The post translational modification of protein occurs through carbamylation. Carbamylation can change the structure of proteins and can modify the biological activity of protein. In the present study, I investigated the effect of carbamylated SEB (CSEB) on the inflammatory response mediated by LPS in HL-60 cells. To determine the anti-inflammatory effect of CSEB, I produced carbamylated SEB using potassium cyanate (KCN) and then examined whether CSEB involved in cytokine releases and apoptosis of LPS-stimulated HL-60 cells. Although CSEB had not any effect on the LPS-stimulated HL-60 cells, the protein levels of IL-8, TNF-${\alpha}$ and IL-$1{\beta}$ were significantly decreased by CSEB without cytotoxicity. CSEB also blocked Akt and NF-${\kappa}B$ activation. These results indicate that the suppressive effect of CSEB in LPS-stimulated cytokine releases is occurred by inhibition of Akt and NF-${\kappa}B$ activity. Through further studies, CSEB may be used as anti-inflammatory molecule that makes the immune system more efficient.

• Journal of Microbiology and Biotechnology
• /
• v.20 no.11
• /
• pp.1513-1520
• /
• 2010

A bacterium capable of producing melanin pigment in the presence of L-tyrosine was isolated from a crop field soil sample and identified as Klebsiella sp. GSK based on morphological, biochemical, and 16S rDNA sequencing. The polymerization of this pigment occurs outside the cell wall, which has a granular structure as melanin ghosts. Chemical characterization of the pigment particles showed then to be acid resistant, alkali soluble, and insoluble in most of the organic solvents and water. The pigment got bleached when subjected to the action of oxidants as well as reductants. This pigment was precipitated with $FeCl_3$, ammoniacal silver nitrate, and potassium ferricynide. The pigment showed high absorbance in the UV region and decreased absorbance when shifted towards the visible region. The melanin pigment was further charecterized by FT-IR and EPR spectroscopies. A key enzyme, 4-hydroxyphenylacetic acid hydroxylase, that catalyzes the formation of melanin pigment by hydroxylation of L-tyrosine was detected in this bacterium. Inhibition studies with specific inhibitors, kojic acid and KCN, proved that melanin is synthesized by the DOPA-melanin pathway.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.17 no.2
• /
• pp.467-475
• /
• 2003

This studt was investigated to prove the effect of GMYM on the brain damage. The results were as follows; 1. GMYM showed significantly inhibitory effect on LDH release by NMDA. AMPA and Kinate. 2. GMYM showed significantly inhibitory effect on LDH release by BSO and Fe2+. 3. GMYM decreased coma duration time in a infatal dose of KCN and showed 30% of survival rate in a fatal dose. 4. GMYM showed improvement of forelimb and hindlimb test after MCA occulusion in neurological exemination. 5. GMYM decreased ischemic area and edema incited by the MCA blood flow block. These results indicate that GMYM can be used in the brain damage sujected to brain ischemia. Further study will be needed about the functional mechanism and etc.

• Korean Journal of Veterinary Research
• /
• v.39 no.1
• /
• pp.70-76
• /
• 1999

Magnesium ($Mg^{2+}$) plays an important role in the regulation of a range of intracellular processes. Regulation of extracellular $Mg^{2+}$ contents was studied in the anesthetized Sprague-Dawley (SD) rats. Animals were injected intraperitoneally with sodium nitrite ($NaNO_2$), and circulating $Mg^{2+}$($[Mg^{2+}]c$) was measured after the injection and then 10 and 20 minutes later. A dose-dependent increase in $[Mg^{2+}]c$ was observed in animals injected with $NaNO_2$ at a dose of 10mg/kg or higher. Pretreatment with methylene blue prevented the $NaNO_2$-induced increase in $[Mg^{2+}]c$. $[Mg^{2+}]c$ displayed an inverse linear correlation with hemoglobin and exponential correlation during $NaNO_2$ injection. Injection of KCN or rotenone also induced an increase in $[Mg^{2+}]c$. An increase in $[Mg^{2+}]c$ was observed when respiration rate was reduced from 100/min (140ml/min) to 10/min (14ml/min) during 30 min. These results indicate that changes in $[Mg^{2+}]c$ inversely reflect alteration of ATP in a model of metabolic inhibition.

• Analytical Science and Technology
• /
• v.8 no.4
• /
• pp.723-730
• /
• 1995

Cyclic voltammetry and in situ STM were employed to examine the interfacial structures of a Pt(111) electrode in 0.1 mM KCN (pH9.5) and 0.1 mM KSCN (pH7) solutions. In situ STM atomic resolution revealed well ordered (2${\surd}$3${\times}$2${\surd}$3)$R30^{\circ}$-6CN and ($2{\times}2$)-2SCN structures within the double layer charging region. Six CN adsorbates formed a hollow hexagon, which embraced a coadsorbed $K^+$ cation. In contrast, the coadsorbed $K^+$ cations on the SCN covered Pt(111) were poorly ordered, despite adsorbed SCN formed a long range ordered ($2{\times}2$)-2SCN adlattice. In situ STM revealed the pronounced influence of potential in controlling the structures of compact layers at the proximity of a Pt electrode. Cathodic polarization facilitated the replacement of the coadsorbed cations by protons.

• The Journal of the Korean Society for Microbiology
• /
• v.22 no.4
• /
• pp.427-433
• /
• 1987

One hundred and thirty-six strains of Klebsiella pneumoniae were isolated from clinical specimen taken from pediatric patiants at 6 different hospitals and identified, characterized and investigated on the patterns of antibiotic resistance. The 136 strains showed the positive reactions in the 17 tests; Voges-Proskauer, ONPG, cirate utilization, KCN degradation, productions of lysine decarboxylase, acid and gas from glucose, utilizations of malonate, manitol, rhamnose, salicin, sucrose, raffinose, arabinose, lactose, sucrose, inositol and raffinose, but the strains showed the negative reactions in the 8 tests; production of $H_2S$, indole, arginine dehydrolase, ornithine decaraoxylase, phenylalanine deaminase, motility, methly red and gelatin liquefaction. 41 did not utilize dulcitol, and 32 did not utilize adonitol. The 36 out of them produced klebecin. The 136 strains were resistant to ampicillin, 2 to gentamicin, 14 to cephalothin, 16 to chloramphenicol, 8 to kanamycin, 13 to streptomycin, and 17 to tetracycline. Twenty strains were resistant to 2 kinds of antibiotics 5 strains to 3, 4 to 4 and 1 to 6 and 7.

• Journal of fish pathology
• /
• v.1 no.2
• /
• pp.95-101
• /
• 1988

During the period from August through October, 1988, 50 isolates of Edwardsiella tarda were isolated from 6 diseased cultured Channa argus in Dunchi island and Myung-ghi, near Pusan in Korea were examined by studying their biochemical and antibiotical reactions. The ill animals moved slowly and irregular-formed swimming at the surface of the corner. The symthoms were necrosis with hemorrhage on the body surface, head, gill region, and mouth. Some fish were observed dropsy of the belly. The bacteria grew slowly on Double Salmonella-Shigella agar, 24h, at $37^{\circ}C$ to form relatively small size (2mm diameter), smoothed and convexed form with transient or black in center of the colonis. They gave negative reactions to Voges-Proskauer, Simmon's citrate, urea, KCN (in growth), gelatin, arginine dehydrolase, phenylalanine deaminase and many sugars. The isolates showed positive reactions to $H_2S$ (in KIA agar), indol, Methyl-Red, motility, lysine and ornithine decarboxylase, and gas from glucose. 8 drugs tested as chloramphenicol, colistin, gentamicin, kanamycin, lincomycin, nalidixic acid spectinomycin, and tetracycline. All cultures were resistant to colistin, lincomycin and spectinomycin respectibly, but sensitive to kanamycin and nalidixic acid. Three strains showed resistance to chloramphenicol and 2 isolates among them were resistant to two drugs(gentamicin and tetracycline), coincidentally.

• Journal of the Korean Chemical Society
• /
• v.31 no.1
• /
• pp.94-97
• /
• 1987

In a converting reaction of $p-bromo-\alpha-diethylaminoacetophenone$ (1a) and $p-bromo-\alpha-piperidinoacetophenone$ (1b) with KCN and $(NH_4)_2CO_3$, to hydantoin, the expected 5-(p-bromophenyl)-5-diethylaminomethylhydantion (2a) and 5-(p-bromophenyl)-5-piperidinomethylhydantoin (2b) were not formed. Diethylaminomethyl group and piperidinomethyl group were eliminated and p,p'-dibromodiphenylhydantil (4), which might be formed through dimerization of 5-(bromophenyl)-hydantoin (3), and p-bromobenzoic acid (5), which was formed through a degradation of 5-(bromophenyl)-hydantoin, were obtained. In the case of $p-bromo-\alpha-morpholinoacetophenone$ (1c), the expected 5-(p-bromophenyl)-5-morpholinomethylhydantoin (2c) was formed.