• Bulletin of the Korean Chemical Society
• /
• v.6 no.5
• /
• pp.312-317
• /
• 1985

${\beta}$-Glucuronidase (EC 3.2.1.31) which hydrolizes D-glucuronate from ${\beta}$-D-glucuronide was purified from rat liver, using ammonium sulfate fractionation, DEAE-cellulose chromatography, Concanavalin-A Sepharose 4B chromatography and gel filtration on Sephadex G-200. This enzyme has the molecular weight of 280,000 daltons by gel filtration and 75,000 daltons by SDS-polyacrylamide gel electrophoresis. As its funtion is reverse of detoxification in the liver, the inhibition of the enzyme was tested with extracts of several food products and medicinal herbs, some are known as anti-cancer agents. Among them, Panax ginseng and Cortnellus shiiake inhibited the enzyme competitively and the $K_1$ values were $9.22 {\times}\;10^{-2}$ and 0.102 mg/ml, respectively. These inhibitors strongly bound to DEAE-cellulose. The negatively charged amino acids, L-aspartate and L-glutamate, inhibited the enzyme, and $K_1$ value of L-aspartate was 0.80 mM. The interaction between ${\beta}$-glucuronidase and p-nitrophenyl-${\beta}$-D-glucuronide was found to involve ionic forces by the effect of ionic strength on the kinetic constant, Vmax/Km. It was inferred from these findings that cationic group at the active center of the enzyme is probably involved in attacking the substrate.

• The Journal of Korean Medicine
• /
• v.16 no.1 s.29
• /
• pp.281-294
• /
• 1995

The effect of Sam Hwa San on the Na-K-ATPase activity was evaluated in microsomal fraction prepared from rabbit cerebral cortex to determine whether Sam Hwa San affects Na-K-ATPase activity of nervous system. Sam Hwa San markedly inhibited the Na-K-ATPase activity in a dose-dependent manner with an estimated $I_{50}$ of 0.12%. Optimal pH for the Na-K-ATPase activity was at 7.5 in the presence or absence of Sam Hwa San. The degree of inhibition by the drug more increased at acidic and alkalic pHs than neutral pH. Kinetic studies of substrate and cationic activation of the enzyme indicate classic noncompetitive inhibition fashion for ATP, Na and K, showing significant reduction in Vmax without a change in Km. Dithiothreitol, a sulfhydryl reducing reagent, partially protects the inhibition of Na-K-ATPase activity by Sam Hwa San. Combination of Sam Hwa San and ouabain showed higher inhibition than cumulative inhibition. These results suggest that Sam Hwa San inhibits Na-K-ATPase activity in central nervous system by reacting with, at least a part, sulfhydryl group and ouabain binding site of the enzyme protein, but with different binding site from those of ATP, Na and K.

• Asian-Australasian Journal of Animal Sciences
• /
• v.14 no.9
• /
• pp.1216-1220
• /
• 2001

Effect of hydroquinone (HQ) on rumen urease activity was studied. Hydroquinone at concentrations of 0.01 ppm, 0.1 ppm, 1 ppm, and 10 ppm inhibited urease activity of intact rumen microbes in vitro by 25%, 34%, 55% and 64% respectively. In the presence of low concentrations of $\beta$-mercaptoethanol, rumen urease could be solubilized and partially purified. The Km for the enzyme was $2{\times}10^{-3}$ M with Vmax of $319.4{\mu}moles/mg$ min. The kinetics of inhibition with partially purified rumen urease was investigated. The result showed that the inhibitory effect was not eliminated by increasing urea concentrations indicating a noncompetitive effect in nature with an inhibition constant $1.2{\times}10^{-5}$ M. Hydroquinone at the concentration of 10 ppm produced 64% urease inhibition, did not affect ruminal total dehydrogenase and proteolytic enzyme (p>0.05), but increased cellulase activity by 28% (p<0.05) in vitro. These results indicated that hydroquinone was a effective inhibitor of rumen urease and could effectively delay urea hydrolysis without a negative effect. The inhibitor appeared to offer a potential to improve nitrogen utilization by ruminants fed diets containing urea.

• Journal of the Korean Society of Tobacco Science
• /
• v.9 no.2
• /
• pp.69-75
• /
• 1987

Nicotine, the main alkaloid of tobacco, showed different effect according to the enzyme. Among investigated enzymes, protease was inactivated remarkably by nicotine and the mode of inhibition was examined. $\alpha$-amylase and $\beta$-amylase were not affected, and cellulase and glucoamylase were inactivated partially when the concentration of it was over 1.0% , but protease was inhibited powerfully by nicotine The inhibition of protease by nicotine was performed almost in the initial stage of reaction, and was not so much affected by temperature, and was reversible. The inhibition type of protease by nicotine appeared as a Mixed-type inhibition.

• Journal of Microbiology and Biotechnology
• /
• v.9 no.4
• /
• pp.414-421
• /
• 1999

Helicobacter pylori were found to be resistant to azide but sensitive to vanadate, suggesting that defect in the P-type ATPase activity rather than F-type ATPase would be lethal to cell survival or growth. To elucidate the relationship between this enzyme inhibition and H. pylori death, we determined the effect of omeprazole (OMP) plus vanadate on enzyme activity and cell growth. The minimum inhibitory concentration (MIC; ca. 0.8$\mu$mol/disk) of vanadate for H. pylori growth was lowered over l0-fold with the aid of OMP, whereby its inhibitory potential toward the P-type ATPase activity was diametrically increased. Alternatively, we found that this enzyme activity was essential for active transport in H. pylori. From these observations, we strongly suggest that the immediate cause of the growth inhibition of H. pylori cells with OMP and/or vanadate might be defective in the cell's active transport due to the lack of P-type ATPase activity. From the spectral data with circular dichroism (CD) spectroscopy, we found that activated OMP (OAS) at concentration below MIC did not disrupt helical structures of membrane proteins. Separately, we determined the cytopathic effect of OAS by SDS-PAGE, indicating the change in the production of cytoplasmic protein but not cell membrane.

• The Journal of Korean Medicine
• /
• v.22 no.2
• /
• pp.3-9
• /
• 2001

Objective : Oriental medicinal plants reported to be used as anti-hypertensive drugs have been in vitro screened for inhibitory effects on angiotensin-converting enzyme (ACE). Methods : The bioassay is based on inhibition of plasma angiotensin-converting enzyme, as measured from the enzymatic cleavage of the Hip-His-Leu substrate into His-Leu. The plant material is extracted with hexane, ethylacetate, n-buthanol and water separately. Results : In total, 51 species (202 extracts) have been investigated and $400{\;}\mu\textrm{g}/ml$ of the solvent extracts from 26 extracts inhibit the enzyme activities by more than 50%. Among them, four samples of two plant species (buthanol and ethylacetate extracts of Salvia miltiorrhiza and buthanol and water extracts of Jeffersonia dubia) were found to posses a high ACE inhibition ability more than 90%. Conclusion : These results suggested that many Oriental medicinal plants have a antihypertensive effects by inhibition of ACE.

• Journal of the Korea Academia-Industrial cooperation Society
• /
• v.17 no.1
• /
• pp.152-158
• /
• 2016

Glutamate is one of the major excitatory neurotransmitters in the central nervous system of vertebrates. Human GDH (hGDH) is the enzyme that regulates the glutamate metabolism and its expression is higher in the brains of schizophrenia patients than in normal subjects. This study examined the changes in the hGDH enzymatic activity caused by antipsychotic drugs (haloperidol, risperidone, (${\pm}$)-sulpride, chlopromazine hydrochloride, melperone, (${\pm}$)butaclamol, domperidone, clozapine) related to schizophrenia. First of all, hGDH isozymes (hGDH1, hGDH2) were synthesized by genetic recombination. As a result of the enzyme assay, haloperidol, (${\pm}$)-sulpride, melperone and clozapine had an inhibitory effect on the hGDH isozymes. In addition, haloperidol showed a non-competitive inhibition against the substrate, 2-oxoglutarate. In contrast, it showed an uncompetitive inhibition against another substrate, NADH. The inhibitory effect of haloperidol on hGDH2 was abolished by the presence of L-leucine, an allosteric effector of hGDH, but by not other antipsychotic drugs. These results revealed the inhibition of enzyme activity by psychotropic drugs in hGDH isoenzymes (hGDH1 and hGDH2) and the possibility that haloperidol may be used to regulate the GDH activity and glutamate concentration in the central nervous system.

• Microbiology and Biotechnology Letters
• /
• v.13 no.3
• /
• pp.173-178
• /
• 1985

Extracellular amylase from tile filtrate of the submerged culture of Ganoderma lucidum was partially purified by ammonium sulfate precipitation and its properties were studied. The optimum pH and temperature of the enzyme activity were 5.5 and 5$0^{\circ}C$. respectively. This enzyme was most stable at pH 5.0 and stable up to 3$0^{\circ}C$, but it lost completely the activity when it was treated at 6$0^{\circ}C$ for 10 min. The enzyme was activated by the addition of M $n^{++}$, $C^{++}$ and C $u^{++}$, but inhibited by H $g^{++}$, A $g^{++}$ And various enzyme inhibitors and chemical reagents did not affect the enzyme activity. The enzyme hydrolyzed the boiled amylaceous polysaccharides, but it hydrolyzed raw starches very slowly. The activation energy of the enzyme for soluble starch was calculated and found to be 7.06 Kcal per mole. The Km values of the enzyme for soluble starch, amylose, amylopectin and glycogen were 0.16, 0.37, 0.19, and 0.16mg/$m\ell$, respectively. Maltose was found to inhibit the enzyme activity and kinetic analysis revealed a competitive type of inhibition.n.n.n.n.n.

• BMB Reports
• /
• v.31 no.3
• /
• pp.296-302
• /
• 1998

A potentiometric flow injection biosensor for the analysis of anti-cholinesterases (anti-ChEs), based on inhibition of enzyme activity, was developed. The sensor system consists of a reactor with acetylcholinesterase (AChE) immobilized on controlled pore glass and a detector with an $H^{+}-selective$ PVC-based membrane electrode. The principle of the analysis is based on the fact that the degree of inhibition of AChE by an anti-ChE is dependent on the concentration of the anti-ChE in contact with AChE. The sensor system was optimized by changing systematically the operating parameters of the sensor to evaluate the effect of the changes on sensor response to ACh. The optimized biosensor was applied to the analysis of paraoxon, an organophosphorus pesticide. Treatment of the inhibited enzyme with pyridine-2-aldoxime fully restored the enzyme activity allowing repeated use of the sensor.

• Korean Journal Plant Pathology
• /
• v.10 no.2
• /
• pp.119-122
• /
• 1994

Effects of organophosphorus fungicides on cutinase activity from Glomerella cingulata causing apple anthracnose and infection of apple were investigated. Diisoprophylfluorophosphate (DFP) inhibited the enzyme activity indicating that catalysis involves an active serine. In inhibition of the enzyme activity by organophosphorus fungicides, I50 (molar concentration of fungicide at which the enzyme activity is inhibited 50%) of Hinosan and Kitazin P were 26.3 $\mu$M and 427.7 $\mu$M, respectively. At concentration of 10-3 M DFP and organophosphorus fungicides, the infection of G. cingulata was inhibited to 5% in comparison with 15% infection at the unwounded healthy control, but increased to 30% when added with 1 mg/ml of cutinase. Mycelial growth was 36 mm in colony diameter on the medium added with 10-4M of hinosan in comparison with 90 mm of the untreated control, but was 90 mm on the medium added with 10-4M of kitazin P showing lower inhibition than hinosan. The spore germination was more than 60% at all the concentrations of both fungicides.