• Title/Summary/Keyword: phosphatase activity

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Gene Cloning, Expression, and Characterization of Glucose-1-Phosphatase from Enterobacter cloacae B11

  • Kim, Young-Ok;Park, In-Suk;Nam, Bo-Hye;Kong, Hee-Jeong;Kim, Woo-Jin;Lee, Sang-Jun;Kim, Kyung-Kil
    • Fisheries and Aquatic Sciences
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    • v.13 no.1
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    • pp.49-55
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    • 2010
  • A bacterial strain with phytase and glucose-1-phosphatase activity was isolated from seawater. The colony was identified as an Enterobacter cloacae strain and named E. cloacae B11. A gene, agpEnB11, coding for an intracellular acid glucose phosphatase was cloned from the strain and sequenced. It comprised 1,242 nucleotides and encoded a polypeptide of 413 amino acids. Recombinant glucose-1-phosphatase (AgpEn) was overexpressed in Escherichia coli and purified using Ni-NTA column under native conditions. Purified protein displayed a single band of 47 kDa on SDS-PAGE. AgpEn hydrolyzed a wide variety of phosphorylated compounds, with high activity for glucose-1-phosphate and glucose-6-phosphate. Optimum pH and temperature for enzyme activity were pH 5.0 and $50^{\circ}C$, respectively. Enzyme activity was stimulated by $Ca^{2+}$ and $Co^{2+}$, and inhibited by $Cu^{2+}$.

Histochemistry of the Developing Chick Respiratory Tract (鷄胎呼吸氣道의 組織化學的 硏究)

  • Hah, Jae Chung
    • The Korean Journal of Zoology
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    • v.15 no.4
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    • pp.183-191
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    • 1972
  • A histochemical study of the respiratory tract in developing chick was done to demonstrate PAS-postivie materials, ribonucleic acid, phospholipid, and alkaline phosphatase activity. Following results are obtained: 1. The alkaline phosphatase activity was found to be high before the appearance of cartilage in mesenchyme surrounding the tracheal epithelium. The enzyme activity declined after the cartilage formation, followed by the restricted activity in epithelium in the postembryonic stage. 2. A moderate positive reaction of ribonucleic acid was found in the cytoplasm of the epithelium and undifferentiated mesenchyme. As the cartilage grew differentiated the reaction of ribonucleic acid was found to disappear in the mesenchyme surrounding the epithelim, but the cytoplasm of the glands showed a moderate positive reaction. 3. Goblet cells of the mucosa and glandular cells showed highly positive reaction, but the basement membrane exhibited slightly positive PAS-reaction. 4. Epithelial cells of the mucosa showed a weak to moderate reaction. However, the epithelia of bronchiol and alveoli in the differentiating period and glandular cells showed a strong positive reaction in Baker's hematein test.

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Effects of Korean Ginseng on the Visceral Alkaline Phosphatase Activity and Blood Inorganic Phosphorus Level (인삼이 장기의 Alkaline Phosphatase활성과 혈중 무기 인량에 미치는 영향)

  • Jung, Noh-Pal
    • The Korean Journal of Physiology
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    • v.7 no.1
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    • pp.1-11
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    • 1973
  • To study the effects of Korean ginseng on mice visceral alkaline phosphatase activity and inorganic phosphate phosphorus level in the blood, the experimental groups of male and female mice were injected subcutaneously with 0.01 ml or 0.02 ml of Tyrode solution per 10 g of body weight daily, and 0. 1 mg or 0.2 mg of alcohol extract of ginseng which was diluted with Tyrode solution. After groups of mice were treated for 7 days or 14 days, the alkaline phosphatase activity in the jejunum, kidney and liver homogenate and serum were determined with sodium ${\beta}-glycerophophosphate$ as substrate, and quantified the content of inorganic phosphate phosphorus in the blood of above mice. 1. Alkaline phosphatase activity in the jejunum, kidney, liver and serum of male mice for the 7 day treatment with ginseng extract (0.1 mg/10 g) was increased by 15.28%, 12.86%, lg.05% and 11.70% respectively, compared with Tyrode solution group. 2. Alkaline phosphatase activity in the jejunum, kidney, liver and serum of female mice for the 7 day treatment with ginseng extract (0.2 mg/10 g) was increased 3.46%, 6.94%, 20.37% and 4.05 % respectively. 3. Alkaline phosphatase activity in the jejunum, kidney, liver and serum of male mice for the 14 day treatment with ginseng extract(0.1 mg/10 g) was increased·15.92%, 19.76f, 10.16% and -1.63 % respectively. 4. Alkaline phosphatase activity in the jejunum, kidney, liver and serum of female mice for the 14 day treatment with ginseng extract(0.2 mg/10 g) was increased 18.89%, 24.55%, 16.97% and 27.59% respectively. 5. Increasing activity of the enzyme in the liver of both male and female mice for the 7 day treatment was declined to some extent at the 14 day treatment, on the other hand, increasing activity of the enzyme in the jejunum, kidney and serum of mice for the 7 day treatment was more promoted at the 14 day treatment. 6. The 7 day and 14 day treatment with ginseng extract increased 20.20% and 20.96% of inorganic phosphate phosphorus in male blood, 22.38% and 17.57f in female blood respectively. In accordance with the results mutual relationships of ginseng, internal secretion, nucleic acid and alkaline phosphatase activity were disscussed.

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The Effects of Phosphate Starvation on the Activities of Acid and Alkaline Phosphatase, Fructose-1,6-bisphosphatase, Sucrose-phosphate Synthase and Nitrate Reductase in Melon (Cucumis melo L.) Seedlings

  • Kang, Sang-Jae;Lee, Chang-Hee;Park, Man
    • Korean Journal of Soil Science and Fertilizer
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    • v.49 no.1
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    • pp.44-52
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    • 2016
  • Plants response to phosphate starvation include the changes of activity of some enzymes, such as phosphatases, fructose-1,6-bisphosphatase, sucrose-phosphate synthase and nitrate reductase. In this study, to determine the effects of phosphate starvation on the change of activities of acid and alkaline phosphatase, fructose-1,6-bisphosphatase, sucrose-phosphate synthase, and nitrate reductase were studied in melon seedlings (Cucumis melo L.). The content of the protein and chlorophyll tended to relatively reduced in melon seedlings subjected to phosphate starvation. Acid phosphatase activity in first and second leaves of melon seedlings was relatively higher than that of third and fourth leaves of seedlings in 14 days after phosphate starvation treatment, respectively. Active native-PAGE band patterns of acid phosphatase in melon leaves showed similar to activities of acid phosphatase, whereas alkaline phosphatase activity was different from the change in the activity of acid phosphatase. Inorganic phosphate content in melon seedlings leaves was constant. The changes of Fructose-1,6-bisphosphatase and sucrose phosphate synthase activities showed similar patterns in melon seedlings leaves, and between these enzymes activities and phosphate nutrition negatively related. Fructose-1,6- bisphosphatase and sucrose phosphate synthase activities showed significant difference in second and fourth leaves, but nitrate reductase showed significant difference in first and second leaves in 14days after phosphate starvation treatment. We concluded that phosphate nutrition could affect the distribution of phosphate, carbon and nitrogen in melon seedlings.

Inhibitory Effect of Schizandrin on Toxicity of Paraquat (Paraquat 독성에 대한 Schizandrin의 억제효과)

  • 정세영
    • Environmental Analysis Health and Toxicology
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    • v.13 no.3_4
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    • pp.117-123
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    • 1998
  • Paraquat is a useful nonselective herbicide widely used throughout the world. However, accidental or intentional ingestion of the paraquat cause fetal pulmonary injuring. But there is not suitable antidote of paraquat intoxication and therapeutic agents now be used are not effective. So, in this study we intended to evaluate the inhibitory effects of DDB(dimethyl-4,4'dimethoxy-5,6,5',6'-dimethylene dioxyphenyl-2,2'-dicarboxylate) on paraquat toxicity. DDB (100mg/kg) was administered orally to SD rats lhr after paraquat(50mg/kg) injection. After 24 hours, the biochemical parameters of blood and tissues were examined. In paraquat treated groups sGPT, sGOT, BUN, creatinine, MDA and alkaline phosphatase levels in blood and MDA, glucose-6-phosphatase activity in tissues were elevated by 2 to 5 times of normal values. However in schizandrin treated groups, sGPT, sGOT, MDA and alkaline phosphatase activity in blood and MDA and glucose-6-phosphatase activity were significantly decreased to notmal levels but not in biochemical parameters of nephrotoxicity, BUN and creatinine levels. Therefore, we concluded that schizandrin can be used as an antidote of pulmono, hepatotoxicity of paraquat.

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A Study of Alkaline Phosphatase Activity on the Preimplantation Mouse Embryos (초기 흰쥐 배아의 발생단계에 있어서의 Alkaline Phosphatase의 활성에 관한 연구)

  • Cho, Wan-Kyoo;Lee, Chung-Choo;Kim, Hee-Kwon
    • The Korean Journal of Zoology
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    • v.27 no.1
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    • pp.1-12
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    • 1984
  • In order to investigate the alkaline phosphtase activities in the mouse oocytes in matuation and preimplantation embryos in developing in culture, the enzyme activities were measured by means of biochemical method. The in vitro effect of levamisole which is known as an inhibitor of the lakaline phosphatase was also observed on the oocyte in maturation and the embryos in early embryogenesis. The results obtained were as follows: The enzyme activity was not detected in the embryos unitl the stage of 4-cell, but it appeared first in the 4-cell embryos and the level of the activity was steady through up to the blastocyst. Levamisole inhibited the alkaline phosphatase activity in the blastocyst, and the activity decreased by almost 70% at 10 mM and 50% at 1 mM as compared with the control. In addition, levamisole inhibited completely the formation of polar body by the oocytes. and induced degeneration of the preimplantation embryos at the dose of 0.5 mM or higher.

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Radiomodifying Potential of Panax ginseng in Liver of Swiss Albino Mice against Gamma Radiation

  • Sharma, Mukesh Kumar;Kumar, Madhu;Kumar, Ashok
    • Journal of Ginseng Research
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    • v.28 no.2
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    • pp.78-86
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    • 2004
  • Panax ginseng occupies an important role in the folk medicine of China, Korea and Japan. The present study was undertaken to determine the radioprotective efficacy of ginseng root extract in the liver of Swiss albino mice. The animals were divided into 4 groups. Group I-Only vehicle was administered. Group II-The animals received 10 mg/kg body weight ginseng root extract i.p. for 4 consecutive days. Group III-Animals were irradiated with 8Gy gamma radiation at the dose rate of 1.69 Gy/min at the distance of 80 ems. Group IV-Animals were given by ginseng root extract (10 mg/kg body weight) continuously for 4 days and on 4th day they were irradiated with 8 Gy gamma radiation after 30 min. The animals from above groups were autopsied on 1,3,7,14 and 30 days. Biochemical estimations of phosphatases (acid & alkaline), LDH (lactate dehydrogenase), LPO (lipid peroxidation) and GSH (reduced glutathione) in liver and SGOT (serum glutamate oxaloacetate transaminase), SGPT (serum glutamate pyruvate transaminase) and alkaline phosphatase in serum were done. In ginseng treated group acid phosphatase (ACP), alkaline phosphatase (ALP), LPO and LDH in liver and SGOT, SGPT and alkaline phosphatase in serum did not show any significant alteration. However, a significant increase in GSH content in liver was recorded. In irradiated group there was a significant increase in ACP, ALP and LPO content in liver and SGOT & SGPT in serum was noted. Whereas, a significant decrease was recorded in GSH and LDH activity in liver and alkaline phosphatase activity in serum. Pretreatment of ginseng with radiation significantly alters the biochemical parameters in liver and serum. A significant decline in ACP, ALP activity and LPO content in liver and SGOT and SGPT activity in serum was observed. However, a significant increase in GSH content and LDH activity in liver and ALP activity in serum was estimated. The present study suggests that pretreatment of ginseng before irradiation significantly protects the liver and maintains the enzyme activity.

THE EFFECTS OF MECHANICAL STRESS ON CULTURED BONE CELL POPULATIONS (Mechanical stress가 골조직세포군에 미치는 영향)

  • Kim, Sang-Tae;Cha, Kyung-Suk
    • The korean journal of orthodontics
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    • v.24 no.1 s.44
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    • pp.105-114
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    • 1994
  • The movement of teeth during orthodontic treatment requires bone remodeling process of bone formation and bone resolution. To find out the changes occuring in the cell itself, mechanical stress was applied to the cell populations involved in the bone metabolism. Bone tissue cell populations were isolated from fetal rat calvaria and divided into OC and OB groups. Following results were obtained from measuring the changes in acid & alkaline phosphatease activity, cyclic AMP and $PGE_2$ production in time lapse after the application of mechanical stress. 1. In case of the marker enzyme of specific bone tissue cell, acid phosphatase activity was high in OC group and alkaline phosphatase activity was high in OB group. 2. After the mechanical stress was applied, acid phosphatase activity was decreased in both OC and OB groups and alkaline phosphatase activity was increase in OB group. 3. When the mechanical stress was applied for 15, 30 and 60 minutes, the production of $PGE_2$ increased in both OC and OB groups, as the time span increased. 4. When the mechanical stress was applied for 20 and 40 minutes, the production of $PGE_2$ increased in both OC and OB groups, as the time span increased.

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Effects of Cyclic-AMP and Tannin on the Amylase Biosynthesis Induced by Gibberellin in Aleurone Layer I. Acid Phosphatase (Cyclic-AMP와 탄닌이 지베레린으로 유도되는 Amylase 생합성에 미치는 영향 I. Acid Phosphatase)

  • 권영명
    • Journal of Plant Biology
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    • v.21 no.1_4
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    • pp.13-21
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    • 1978
  • The effect of cyclic-AMP on the induction of acid phosphatase activity in barley aleurone layers was examined. Tannic acid was used as a inhibitor. Decursinol and coumarin were also used as a comparison. Maxiumu promotion of the enzyme activity was obtained with 10-5M cyclic-AMP, but this promotion was lower than that of 10-5M GAS induced enzyme activity in incubation medium. The inhibition rate in the addition of tannic acid was shown 17% and 63% at a ratio to GAs (by weight) of 10 : 1, and 58% and 94% at a ratio of 100 : 1 treated with GAs, and cyclic-AMP, respectively. The most potentiation of 10-6M GAS effect was induced by the additiion of suboptimal concentration (10-6M) of cyclic-AMP. Additional GAs and cyclic-AMP were shown the recovery of the enzyme activity inhibited by tannic acid. The combination with cyclic-AMP and theophylline enhanced the enzyme activity, too. Any other nucleotides tested except cyclic-AMP didn't show the action. There were no differences in acid phosphatase isozyme patterns by polyacrylamide disc electrophoresis, in conjunction with the different additions but the size of bands showed great differences. Especially, the 3rd band and the 5th band group were remarkable.

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Chemical Modification of Porcine Brain myo-Inositol Monophosphate Phosphatase by N-bromosuccinimide

  • Lee, Byung-Ryong;Bahn, Jae-Hoon;Jeon, Seong-Gyu;Ahn, Yoon-Kyung;Yoon, Byung-Hak;Kwon, Hyeok-Yil;Kwon, Oh-Shin;Choi, Soo-Young
    • BMB Reports
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    • v.32 no.3
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    • pp.294-298
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    • 1999
  • Myo-inositol monophosphate phosphatase is a key enzyme in the phosphoinositide cell-signaling system. Incubation of myo-inositol monophosphate phosphatase from porcine brain with N-bromosuccinimide (NBS) resulted in a time-dependent loss of enzyme activity. The inactivation followed pseudo-first-order kinetics with the second-order rate constant of $3.8{\times}10^3\;M^{-1}min^{-1}$. The time course of the reaction was significantly affected by the substrate myo-inositol-1-phosphate, which afforded complete protection against the loss of catalytic activity. Spectrophotometric studies indicated that about one oxindole group per molecule of enzyme was formed following complete loss of enzymatic activity. It is suggested that the catalytic function of myo-inositol monophosphate phosphatase is modulated by the binding of NBS to a specific tryptophan residue at or near the substrate binding site of the enzyme.

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