• Title/Summary/Keyword: Ca-EDTA

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Purification and Characterization of a Maltopentaose-producing Amylase from Bacillus megaterium KSM B-404. (Bacillus megaterium KSM B-404으로부터 생산되는 Maltopentaose생성 Amylase의 정제 및 특성)

  • 박제원;김병주;이재우;김영배
    • Microbiology and Biotechnology Letters
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    • v.30 no.4
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    • pp.352-358
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    • 2002
  • An amylase that hydrolyzes starch into maltopentaose as a main product was found in the culture supernatant of a strain of Bacillus megaterium KSM B-404 isolated from local soil. The enzyme was purified 129-fold by ammonium sulfate precipitation, DEAE-Toyopearl and Superdex 75 HR 10/30 column using a FPLC system. The molecular weight of the amylase was determined as about 68 kDa by using SDS-PAGE. Optimum pH and temperature of amylase were found to be $50^{\circ}C$ and pH 6.0~7.0, respectively. The enzyme was stable up to $60^{\circ}C$ by addition of $Ca^{2+}$ and its pH stability was in the range of 6.0~10.0. The activity of enzyme was inhibited by $Cu^{2+}$ $Hg^{2+}$ , and $Fe^{3+}$ and maintained by $Ca^{2+}$ and $Mg^{2+}$ . EDTA and pCMB also showed inhibitory effect to the enzyme. TLC and HPLC analysis of the products of the enzyme reaction showed the presence of maltopentaose(52%), maltotriose (25%), maltose (11%), glucose, and maltotetraose in the starch hydrolysates.

Hydrothermal Coating of Hydroxyapatite on ZrO2 Ceramics

  • Ha, Jung-Soo
    • Journal of the Korean Ceramic Society
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    • v.43 no.8 s.291
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    • pp.463-468
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    • 2006
  • Hydrothermal deposition of hydroxyapatite coatings on two types of $ZrO_2$ substrates (3 mol% $Y_2O_3$-doped and 13 mol% $CeO_2$-doped tetragonal $ZrO_2s$) was studied using aqueous solutions of $Ca(NO_3)_2\;4H_2O$ and $(NH_4)_2HPO_4$ containing EDTA (ethylene diamine tetra acetic acid) disodium salt as a chelating agent for $Ca^{2+}$ ions. For the precipitation of the coatings, the $EDTA-Ca^{2+}$ chelates were decomposed by oxidation with $H_2O_2$ at $90^{\circ}C$. The deposition behavior, morphology, and orientation of the coatings were investigated while varying the solution pH using scanning electron microscopy and X-ray diffractometry. For the two sub-strates, sparse deposition of the coating was obtained at pH 5.5, whereas a uniform deposition was obtained at pH 7.1, 9.8, and 11.4 with a denser microstructure for the higher pH. The coating consisted of thin needle-like or plate-like crystals ($1-2{\mu}m$ length or diameter) at pH 7.1, but fine rod-like crystals ($1-2{\mu}m$ length, $0.1{\mu}m$ diameter) at pH 9.8 and 11.4. The coatings were $1-3{\mu}m$ thick and showed a preferred orientation of the hydroxyapatite crystals with their c axis (i.e., the elongated direction) perpendicular to the substrate surface especially for pH 9.8 and 11.4.

Identification of a Proper Phytoavailable Arsenic Extraction Method Associated with Arsenic Concentration in Edible Part of three Crops in Soils Near Abandoned Mining Areas

  • Yoon, Jung-Hwan;Kim, Young-Nam;Lee, Dan-Bi;Kim, Kwon-Rae;Kim, Won-Il;Kim, Kye-Hoon
    • Korean Journal of Soil Science and Fertilizer
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    • v.50 no.6
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    • pp.497-508
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    • 2017
  • This study aimed to investigate correlations between concentrations of extractable Arsenic (As) with varying chemical solutions (0.1 M $Ca(NO_3)_2$, 0.1 M $(NH_4)2HPO_4$, 0.5 M EDTA, Mehlich 3, and 0.5 M $NaHCO_3$) and those of As in crops, and then to seek the most suitable soil extraction method for predicting the potential of As uptake in crops cultivated in soils contaminated with As. For a mesocosm experiment, pepper (Capsicum annuum L.), soybean (Glycine max L.), and rice (Oryza sativa L.) were cultivated for three months in pots containing soils taken from the arable areas near abandoned mines in Korea. Following the cultivation, soil pH and DOC significantly increased by treatments of lime and lime plus compost, respectively, while insignificant influences in changing total and all extractable As concentrations were found in all soils. Arsenic concentration in edible part of all crops considerably depended on the extractable As concentration in the soils, particularly with Mehlich 3. All extractable As concentrations in the soils of C. annuum and G. max were significantly correlated with As concentration in their edible parts. For O. sativa, the extractable concentrations of Mehlich 3 ($R^2$: 0.18 at p: 0.006) and EDTA ($R^2$: 0.11 at p: 0.036) showed only marked relationships with As concentration in the edible part. These results may indicate that the Mehlich 3 and EDTA are soil extractants to determine phytoavailable As in soil that provide better prediction for As transfer from soil to crop.

Metal Protease from Streptomyces spp. - I. Isolation of the Strain and the Enzymatic Properties - (Streptomyces 속균(屬菌)이 생성하는 Metal Protease - 제 1 보 : 균(菌)의 분리(分離) 및 효소학적(酵素學的) 성질(成質) -)

  • Yi, Dong-Heui;Yu, Choon-Bal
    • Korean Journal of Food Science and Technology
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    • v.12 no.1
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    • pp.13-17
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    • 1980
  • A Streptomyces spp. strain SY 79-1 which was capable of producing metal protease was isolated from soil. The optimal pH and temperature of the protease were around pH 8.0 and $45^{\circ}C$, respectively. The stable pH range of the enzyme was between pH 6.0 to 8.0. The enzyme was stable at $45^{\circ}C$, but it lost the activity about 75 % for 5 min and completely for 30 min when it was treated at $60^{\circ}C$. The activity of the enzyme was inhibited by $Hg^{++},\;Cu^{++},\;Ag^{+}$ and activated by $Mg^{++},\;Mn^{++},\;Co^{++},\;but\;Fe^{++},\;Ca^{++},\;Pb^{++}\;and\;Al^{3+}$ did not affect enzyme activity. This enzyme was strongly inhibited by EDTA, but was not inhibited by 2, 4-DNP, ${\rho}$-CMB, ${\varepsilon}$-aminocaproic acid, cysteine, thiourea, citric acid, oxalic acid and sodium arsenate. When cobalt was added to the EDTA-denatured enzyme, the activity of the enzyme was restored.

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Comparison of Soil Extractants for Estimation of Cadmium, Zinc and Lead in Brown Rice Collected from Paddy Soils near Old Zinc-Mining Sites II. A prediction Model for Cadmium, Zinc and Lead Contents in Brown Rice Based on Some Chemical Properties of Soils (현미중(玄米中) 중금속(重金屬) 함량예측(含量豫測)을 위한 토양침출액(土壤浸出液)의 비교(比較) II. 토양분석(土壤分析)에 의(依)한 현미중(玄米中) 중금속(重金屬) 함량(含量) 예측(豫測))

  • Yoo, Sun-Ho;Park, Moo-Eon
    • Korean Journal of Environmental Agriculture
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    • v.4 no.1
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    • pp.31-36
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    • 1985
  • In order to choose a suitable soil extractant for the prediction model of heavy metal content in brown rice, four extractants-0.1 M HCl, 0.1 M $HNO_3$, 0.1 M $NH_4$-oxalate and 0.001 M 2Na-EDTA, were compared by analyzing 84 soil and 45 brown rice samples collected from paddy fields adj-acent to five old zinc-mining sites. Content of cadmium and zinc in brown rice had the highest correlation coefficient to 0.001 M 2Na-EDTA and 0.1 M HCl extractants, respectively. However, the lead content in brown rice was significantly correlated with only 0.1 M $NH_4$oxalate solution. For the simultaneous prediction of zinc, cadmium and lead in brown rice, 0.1 M $NH_4$-oxalate solution was the most effective. On the multiple analysis by using various chemical characteristics of soils, pH and calcium content of soils were effective variables for the estimation of cadmium content in brown rice, while CEC and magnesium content were more effective for the estimation of zinc content in brown rice. Furthermore, for the estimation of lead content in brown rice, factors such as pH, CEC, calcium, magnesium, potassium and organic matter content were important variables in the multiple regression equation.

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Studies on distribution of calcium and electromyogram in experimental hypocalcemia (재래산양의 Hypocalcemia에 있어서의 Ca의 동태 및 근전도학적인 연구)

  • Park Moo Hyen;Kang Chung Boo
    • Journal of the korean veterinary medical association
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    • v.17 no.2
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    • pp.63-71
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    • 1981
  • Experimental hypocalcemia was induced in normal goats by intravenous administration with various concentration of $Na_2-EDTA$ solution. Tile results are summarized as follows: 1. Progressive depression of reflexes and body temperature, paresis,

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Nucleotide Sequence, Structural Investigation and Homology Modeling Studies of a Ca2+-independent α-amylase with Acidic pH-profile

  • Sajedi, Reza Hassan;Taghdir, Majid;Naderi-Manesh, Hossein;Khajeh, Khosro;Ranjbar, Bijan
    • BMB Reports
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    • v.40 no.3
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    • pp.315-324
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    • 2007
  • The novel $\alpha$-amylase purified from locally isolated strain, Bacillus sp. KR-8104, (KRA) (Enzyme Microb Technol; 2005; 36: 666-671) is active in a wide range of pH. The enzyme maximum activity is at pH 4.0 and it retains 90% of activity at pH 3.5. The irreversible thermoinactivation patterns of KRA and the enzyme activity are not changed in the presence and absence of $Ca^{2+}$ and EDTA. Therefore, KRA acts as a $Ca^{2+}$-independent enzyme. Based on circular dichroism (CD) data from thermal unfolding of the enzyme recorded at 222 nm, addition of $Ca^{2+}$ and EDTA similar to its irreversible thermoinactivation, does not influence the thermal denaturation of the enzyme and its Tm. The amino acid sequence of KRA was obtained from the nucleotide sequencing of PCR products of encoding gene. The deduced amino acid sequence of the enzyme revealed a very high sequence homology to Bacillus amyloliquefaciens (BAA) (85% identity, 90% similarity) and Bacillus licheniformis $\alpha$-amylases (BLA) (81% identity, 88% similarity). To elucidate and understand these characteristics of the $\alpha$-amylase, a model of 3D structure of KRA was constructed using the crystal structure of the mutant of BLA as the platform and refined with a molecular dynamics (MD) simulation program. Interestingly enough, there is only one amino acid substitution for KRA in comparison with BLA and BAA in the region involved in the calcium-binding sites. On the other hand, there are many amino acid differences between BLA and KRA at the interface of A and B domains and around the metal triad and active site area. These alterations could have a role in stabilizing the native structure of the loop in the active site cleft and maintenance and stabilization of the putative metal triad-binding site. The amino acid differences at the active site cleft and around the catalytic residues might affect their pKa values and consequently shift its pH profile. In addition, the intrinsic fluorescence intensity of the enzyme at 350 nm does not show considerable change at pH 3.5-7.0.

Purification and Biochemical Properties of Extracellular Phospholipase $A_1$ from Serratia sp. MK1

  • Kim, Myung-Kee;Rhee, Joon-Shick
    • Journal of Microbiology and Biotechnology
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    • v.6 no.6
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    • pp.407-413
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    • 1996
  • A novel type of extracellular phospholipase $A_1$ was isolated from Serratia sp. MK1 and purified to homogeneity by ammonium sulfate precipitation, anion exchange and gel filtration chromatography. The purified enzyme was a monomer with a molecular mass of about 43, 000 Da. This enzyme showed the highest lipolytic activity toward phosphatidylserine among the phosphoglycerides tested, and preferentially catalyzed the hydrolysis of the ester bond in phosphatidic acid to lyso-phosphatidic acid. Enzyme activity was completely inhibited by the addition of a chelating agent such as EDTA, and inhibited enzyme activity was fully recovered by the presence of $Ca^{2+}$. This implies that the enzyme requires $Ca^{2+}$ for activity. The enzyme was stable up to $70^{\circ}C$ when incubated for 1 h at pH 8.5, and the optimal pH and temperature were 8.5 and $50^{\circ}C$, respectively.

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Studies on the Water Quality of the Han River Water, and Water Quality Standards (한강의 수질과 수질규준에 관한 고찰)

  • Choe, Sang
    • 한국해양학회지
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    • v.3 no.2
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    • pp.47-54
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    • 1968
  • Monthly mean values for EDTA hardness in the Han River water were determined for March 1966 through May 1967. The hardness was retained a range of 22-59 mg/L the maximum value of 59 mg/L was approached in the months of mid-winter, indicating a seasonal variation. The annual ranges of Ca and Mg were 3.7-9.1 mg/L, 1.5-10.4 mg/L, respectively. The annual range of Mg:Ca ratio was 0.18-2.60. And the hardness correlated closely with Mg volume. In Korea, fortunately, most river waters are not yet suffering from damaging pollution. We must try to establish our water quality criteria based on scientific data, and make research recommendations for the following water uses; public water supplies, aquatic life and wildlife, industry, recreation and aesthetics.

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Comparisons of Simple Extraction Methods and Availability for Heavy Metals in Paddy Soils (토양 중금속의 단일침출방법과 유효도 비교)

  • Jung, Goo-Bok;Kim, Won-Il;Moon, Kwang-Hyun;Ryu, In-Soo
    • Korean Journal of Environmental Agriculture
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    • v.19 no.4
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    • pp.314-318
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    • 2000
  • To compare heavy metal phytoavailability in paddy soils near five abandoned mining areas, 4 different soil extractants such as 0.1M-HCl, $0.1M-HNO_3$, 0.05M-EDTA, and 0.005M-DTPA were used. Total acid digestion method $(H_2SO_4:HClO_4:HNO_3)$ was also employed to analyze heavy metal content in 30 paddy soils and brown rice. The rates of extracted heavy metal to total content were in the range of $12.1{\sim}39.1%$ for Cd, $20.5{\sim}45.5%$ for Cu, $10.6{\sim}30.7%$ for Pb, and $6.7{\sim}13.0%$ for Zn. 0.1M-HCl and $0.1M-HNO_3$ extractable both Cu and Pb were relatively less extracted at the high soil pH and extractable calcium site(Mine D) whereas 0.05M-EDTA and 0.005M-DTPA extractable Pb were strongly extracted at the same soils. In case of Cd, Cu, and Zn in soil, 4 types of extractable heavy metals and total content were highly correlated with each other. However, there were positive correlations between 0.1 M-HCl and $0.1M-HNO_3$ extractable Pb as well as between 0.05M-EDTA and 0.005M-DTPA extractable Pb, which were relatively similar extractants in chemical properties. The rates of heavy metals in brown rice to total contents in soils were in the order Zn>Cd>Cu>Pb. Specially, the rate of Cd, Pb, and Zn were lower at the highest level of soil pH and Ex. Ca. Both Cd and Zn in brown rice were positively correlated with those of all soil extractants. It was estimated that the solubility following to the plant uptake of Cd and Zn were higher than those of Cu and Pb considering relationships between all kinds of heavy metal contents in soil and those in brown rice.

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