• The Korean Journal of Physiology
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• v.25 no.2
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• pp.159-170
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• 1991

In the rabbit renal artery, mechanisms of contraction by sodium depletion were investigated. The helical strips of isolated renal artery were immersed in the Tris-buffered salt solution. The contractions were recorded isometrically using a strain-gauge transducer. Na-free solution (Na was substituted by Li, choline or sucrose) produced contractions which were dependent on the nature of the Na substitutes. Na-free solution (choline) produced the contraction in ouabain-pretreated artery (Na loaded artery) even in the presence of verapamil. The amplitude of the contraction was dependent on the duration of the pretreatment with ouabain $(10\;^5M)$. Monensin potentiated the effect of ouabain on the contraction. Removal of Ca from bathing solution abolished the contraction and the substitution of Sr for Ca produced the contraction. Divalent cations such as Mg, Mn blocked the depolarization-induced contraction, while they had little effect on the Na-free contraction in Na loaded artery. These results suggest that the contraction induced by Na removal is dependent on the cellular Na content and may be caused by Ca influx via the Na-Ca exchange carrier.

• Journal of Plant Biology
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• v.32 no.4
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• pp.265-274
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• 1989

The effect of Ca2+ on auxin-induced ehtylene production in etiolated mungbean (Vigna radiata W.) hypocotyls was studied. Auxin-induced ethylene production by mungbean seedlings which had been germinated in the presence of 5-10mM Ca2+ (High Ca2+ ; HC) is greater than that by seedlings which had been germinated in distilled water (Low Ca2+ ; LC). The effect of Ca2+ on auxin-induced ethylene production was greatly increased after 12hr of incubation period. The stimulation of auxin-induced ethylene production by Ca2+ was specific, since divalent cations, such as Mg2+ and Mn2+ did not enhance auxin-induced ethylene production. Calcium also promoted ethylene evoluation induced by methionine and 1-Aminocyclopropane-1-carboxylic acid(ACC). The effect of Ca2+ on auxin-induced ethylene production was not caused by increase in free IAA or ACC contents of hypocotyl tissue. Dimethyl sulfoxide and Triton X-100, that disrupts the emembranes, inhibited ethylene production to a greater extent in LC segments than in HC segments. Addition of Ca2+ to the incubation medium for LC segments resulted in enchancement of ethylene production probalby because the membrane integrity is supported under these conditions. Comparison of activity of Ethylene Forming Enzyme(EFE) in LC and HC hypocotyl segments indicated that the enzyme activity of HC was about 2 times higher than that of L.C. It is suggested that Ca2+ increases the activity of plasma membrane-bound EFE through its stabilizing effect onn the membrane, which in turn brings about promotion of ethylene production.

• Journal of Microbiology
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• v.35 no.2
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• pp.134-140
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• 1997

Botrytis cinerea T91-1 has been shown to produce at least four different polygalacturonases into a liquid medium containing citrus pectin, a carbon sousrce. One of the enzymes, which had an apparent molecular weight of 66 kDa estimated by denatured polyacrylamide gel electrophoresis, was purified to electrophoretic homogeneity by a series of procedures including a cetone precipitation, ion exchange, heparin affinity, and reverse phase column chromatographies. The molecular weight of native enzyme was determined to be 64 kDa by gel permeation chromatography indicating the enzyme to be a single polypeptide chain. By viscometric analysis, the enzyme was revealed as exo-polygalacturonase. The enzyme activity was inhibited by divalent cations such as $Ca^{2+}$, $Mg^{2+}$, and Cu$^{2+}$. Km and Vmax for polygalacturonic acid hydrolysis were 0.33 mg/ml and 28.6 nM/min, respectively. The optimum temperature for enzymatic activity was 5$0^{\circ}C$. And the enzyme showed optimal pH values between 4.0 and 5.0. The enzyme was stable upto 12 hours in the range of pH 3 to 8 and at temperature below 3$0^{\circ}C$.

• Journal of Soil and Groundwater Environment
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• v.16 no.5
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• pp.18-23
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• 2011

The electrokinetic transport characteristics of salts were investigated using nitrate and sulfate accumulated saline greenhouse soil. Within 8 days, 95% of nitrate was removed from the soil, while sulfate removal was 19% for 8 days. The low removal of sulfate came from adsorption reaction on the soil particles or organic matter and precipitation with calcium. Divalent cations such as calcium and magnesium were transported toward cathode via electromigration, and most monovalent cation such as potassium was removed. The pattern of residual electrical conductivity was similar with that of sulfate. Based on the results, electrokinetic technique is effective to restore nitrate-accumulated saline soil, but is not effective to restore sulfate-accumulated soil.

• BMB Reports
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• v.29 no.4
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• pp.335-342
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• 1996

Porcine liver cysteinesulfinic acid decarboxylase was purified approximately 460-fold by means of ammonium sulfate fractionation and sequential column chromatographic separation with Sephadex G-100, DEAE-cellulose and hydroxylapatite. The enzyme has a flat pH profile with maximum activity occurring between pH 6.0 and 7.6. Pyridoxal 5'-phosphate must be present in all buffers used for purification procedures in order to stabilize the enzyme. Addition of sulfhydryl reagents such as 2-mercaptoethanol are also necessary to maintain maximum enzyme activity throughout purification. The absorption spectrum shows that cysteinesulfinic acid decarboxylase is a pyridoxal 5' -phosphate-containing protein. The major absorption is at 280 nm with two smaller absorption regions, one at 425 nm which is ascribed to a Schiffs base between pyridoxal phosphate and protein, and another at 325 nm which is thought to be due to the interaction of 2-mercaptoethanol with the Schiffs base. A number of divalent cations tested did not affect enzyme activity with the exception of mercury, copper, and zinc which are inhibitory. The partially purified enzyme has an apparent $K_m$ of 0.94 mM for cysteinesulfinate. Cysteic acid is a competitive inhibitor of the enzyme with a $K_i$ of 1.32 mM. The molecular weight of the enzyme was estimated to be about 79,600 by using Sephadex G-200 column chromatography.

• Journal of Microbiology
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• v.43 no.3
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• pp.237-243
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• 2005

The protease purified from Bacillus cereus JH108 has the function of leucine specific endopeptidase. When measured by hydrolysis of synthetic substrate (N-succinyl-Ala-Ala-Pro-Leu-p-nitroanilide), the enzyme activity exhibited optimal activity at pH 9.0, $60^{\circ}C$. The endopeptidase activity was stimulated by $Ca^{++},\;Co^{++},\;Mn^{++},\;Mg^{++},\;and\;Ni^{++}$, and was inhibited by metal chelating agents such as EDTA, 1,10-phenanthroline, and EGTA. Addition of serine protease inhibitor, PMSF, resulted in the elimination of the activity. The endopeptidase activity was fully recovered from the inhibition of EDTA by the addition of 1 mM $Ca^{++}$, and was partially restored by $Co^{++}\;and\;Mn^{++}$, indicating that the enzyme was stabilized and activated by divalent cations and has a serine residue at the active site. Addition of $Ca^{++}$ increased the pH and heat stability of endopeptidase activity. These results show that endopeptidase requires calcium ions for activity and/or stability. A Lineweaver-Burk plot analysis indicated that the $K_m$ value of endopeptidase is 0.315 mM and $V_{max}$ is 0.222 ) is $0.222\;{\mu}mol$ of N-succinyl-Ala-Ala-Pro-Leu-p-nitroanilide per min. Bestatin was shown to act as a competitive inhibitor to the endopeptidase activity.

• Journal of Microbiology and Biotechnology
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• v.14 no.4
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• pp.647-652
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• 2004

Thermophilic microorganisms capable of producing chitinase enzymes were screened from samples collected from several crater and geothermal areas. The chitinolytic microorganisms were grown in a selective medium containing colloidal chitin. The Bacillus K29-14 isolate was found to exhibit the highest chitinase and chitin deacetylase activities. When grown in a chitin-containing medium, the isolate produced extracellular chitinase after 24 h of incubation. The optimum temperature and pH for the chitinase were $55^\circ{C}$ and pH 7, respectively, while those for the chitin deacetylase were $55^\circ{C}$ and pH 8, respectively. The thermostable chitinase and chitin deacetylase also retained 80- 90% of their activity after incubation for 5 h at $70^\circ{C}$. The divalent cations $CoCl_2\;and\;NiCl_2$, increased the chitinase activity, while $ZnCl_2$, inhibited the enzyme. The chitin deacetylase was also activated by the presence of $MgCl_2$ and inhibited by $MnCl_2,\;NiCl_2,\;and\;CaCl_2$. A zymogram analysis revealed several forms of chitinase, with a 67 kDa form being the major enzyme.

• Economic and Environmental Geology
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• v.21 no.2
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• pp.175-184
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• 1988

Ten under 2 micron size fractions of the montmorillonite from Yongdongri area, Gyeongsangbug-Do were studied using X-ray powder diffraction, cation exchange measurement, differential thermal analysis, thermogravimetric analysis, differential thermal scanning calorimetry and chemical analysis. Montmorillonites occurring at same deposit show limited variation in chemical composition whereas in thermal properties they do not. Their dehydroxylation endothermic peaks are "abnormal" type with a small range of variation of peak temperature reflecting tetrahedral substitution of Al for Si. Data from DSC show that divalent-cation saturated montmorillonite has relatively a higher endothermic heat capacity than monovalent-cation saturated montmorillonite, indicating that cations with higher electronegativity hold more water molecules.

• YAKHAK HOEJI
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• v.33 no.6
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• pp.319-323
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• 1989

Polymorphonuclear leukocyte (PMNL) was obtained from peritoneal cavity in rat treated with casein. Effects of divalent cations and drugs on leukotriene $B_4(LTB_4)$ formation from arachidonic acid in the PMNL were determined by HPLC assay. 5-Lipoxygenase, a key enzyme for $LTB_4$ formation from arachidonic acid, exhibited $V_{max}$ at $1\;{\times}\;10^{-5}M$, and $K_m$ at $9.89\;{\times}\;10^{-5}M$ of arachidonic acid. Optimal $Ca^{++}$ concentration for the enzyme activity was $1\;{\times}\;10^{-5}M$ but $Zn^{++}$ did not show any significant effects on the $LTB_4$ formation. Indomethacin and caffeic acid exhibited inhibitory effects on the $LTB_4$ formation at $1\;{\times}\;10^{-5}M$ and $4\;{\times}\;10^{-5}M$, respectively. However, 6-aminohexanoic acid did not show any significant effects on the $LTB_4$ formation.

• Korean Journal of Microbiology
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• v.27 no.3
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• pp.237-244
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• 1989

A soluble intracellular protease from cells of Pseudomonas carboxydovorans, a carboxydobacterium, grown on nutrient broth was purified 68-fold in five steps to better than 95% homogeneity with a yield of 2.4% using azocasein as a substrate. The enzyme activity was not detected from cells grown on pyruvate, succinate, acetate, or CO as a sole source of carbon and energy. The molecular weight of the native enzyme was determined to be 53,000. Sodium dodecyl sulfate-gel electrophoresis revealed the purified enzyme a monomer. The enzyme was found to be a serine-type protease. The enzyme activity was inhibited completely by several divalent cations such as $Cd^{2+}, Cu^{2+}, Hg^{2+}$, and $Fe^{2+}$. The enzyme was also inhibited by EGTA, but was stimulated by iodoacetamide. The optimal pH and temperature for the enzyme reaction were found to be 8.0 and $50^{\circ}C$, respectively. The enzyme was inactive on CO dehydrogenase.