• Title/Summary/Keyword: Ammonium-exchange

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Purification and Characterization of Aryl Acylamidase from Pseudomonas sp. (Pseudomonas sp. Aryl Acylamidase의 정제 및 성질)

  • 황인균;방원기
    • Microbiology and Biotechnology Letters
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    • v.26 no.5
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    • pp.413-419
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    • 1998
  • Aryl acylamidase [EC 3.5.1.13] present in an acetaminophen-assimilating Pseudomonas sp. has been purified to a homogeneity using series of ammonium sulfate fractionation, DEAE-Sephacel anion exchange, Phenyl-Sepharose CL-4B hydrophobic, and Sephadex G-100 gel-permeation chromatography. The molecular weight, which was estimated by gel-permeation filtration and sodium dodecyl sulfate polyacylamide gel electrophoresis, was about 57 kDa and 56 kDa, respectively, indicating that this enzyme is a monomeric protein. The optimum pH was 10.5 and the optimum temperature was 40$^{\circ}C$. After incubation of the enzyme at 50$^{\circ}C$ for 30 min, residual activity of the enzyme was 34% compared to its original activity. The Km values for acetaminophen and 4'-nitroactanilide were 0.10 mM and 0.11 mM, respectively.

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Purification and Characterization of an Extracellular Xylanase of Bacillus stearothermophilus (Bacillus stearothermophilus 가 생산하는 Xylanase의 정제 및 특성)

  • 배성호;최용진
    • Microbiology and Biotechnology Letters
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    • v.19 no.6
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    • pp.592-597
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    • 1991
  • An extracellular xylanase of Bacillus stearothemophilus was purified to a single protein through a sequency of operations including ammonium sulfate fractionation, DEAE Sepharose CL-6B ion exchange chromatography, Sephadex G-100 gel filtration and heat treatment. The purified enzyme had a moleular weight of 170, 000. the pH and temperature optima for the enzyme activity were pH 9.0 and $55^{\circ}C$, respectively. The activity was enhanced by $co^{2+} \; and\; Mn^{2+}$, and inhibited by $Hg^{2+}$. Pattern of hydrolysis demonstrated that the xylanase was an endo-splitting enzyme able to break down larchwood xylan at random giving xylobiose and xylotriose as the main end products.

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Enzymatic Properties of Serratia marcescens Pretense (Serratia marcescens Protease의 효소학적 특성)

  • 최병범
    • The Korean Journal of Food And Nutrition
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    • v.16 no.2
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    • pp.152-157
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    • 2003
  • Serratia marcescens ATCC 25419 protease was purified to homogeneity by ammonium sulfate treatment, and DEAE-cellulose anion exchange chromatography. The specific activity of the enzyme was increased 448-fold during purification with an overall yield of 43.0%. Metal reactivation on the purified protease from S. marcescens was studied. S. marcescens protease was a metalloenzyme to be completely inhibited its activity by EDTA and the enzyme outstandingly inhibited by Hg, Fe, Cu, but the activity was increased approximately 20% by Co. The reactivation of the apoenzyme was effective with Mn, Co, Zn in pH range from 6 to 8. Among metalloenzymes prepared to the addition of Mn, Co, Zn to restore the degree of activity of native enzyme, Zn-enzyme was similar to the native enzyme in respects with enzyme activity, alkali-inactivation, thermo-stability.

Purification and Properties of $\alpha$-Glucosidase from Mococcus halophilus (Pediococcus halophilus로부터 생성한 $\alpha$-Glucosidase의 정제 및 특성)

  • 민해기;이호근;문지웅;강국희
    • Microbiology and Biotechnology Letters
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    • v.20 no.2
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    • pp.143-149
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    • 1992
  • A bacterial strain No. 2, which highly produced a-glucosidase, was isolated from Kimchi and identified to be a similar species of Pediococcus halophilus. This enzyme was purified by protamine sulfate, ammonium sulfate fractionation, ion exchange and gel filtration. The maximal a-glucosidase activity was observed at pH 6.0 and this enzyme was stable at pH 6.0~ 7.5. The optimum temperature of this enzyme activity was $37^{\circ}C$, but enzyme activity was gradually lost above $37^{\circ}C$. This enzyme was activated by 10 mM MgCh and inhibited by 10 mM mercaptoethanol. The kinetics of PNPG(p-Nitrophenyl-a-D-glucopyranoside) and maltose were Kp0.52 mM/27.5 pg protein, $V_{max}$= 0.021 mM/min 27.5 ${\mu}g$ protein and $K_m$= 0.32 mMD7.5 ${\mu}g$ protein, $V_{max}$= 0.025 mM/min 27.5 ${\mu}g$ protein, respectively. The molecular weight of $\alpha$-glucosidase was about 37, 000.

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Studies on Production of $\beta$-Galactosidase by Lactobacillus sporogenes - Purification of Extracellular $\beta$-Galactosidase - (Lactobacillus sporogenes에 의한 $\beta$-Galactosidase 생산에 관한 연구 -균체외 $\beta$-Galactosidase의 정제 -)

  • 김영만;이정치;최용진;양한철
    • Microbiology and Biotechnology Letters
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    • v.13 no.3
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    • pp.185-189
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    • 1985
  • Extracellular $\beta$-galactosidase from the culture broth of L. sporogenes was purified to apparent homogeniety by procedures including ammonium sulfate fractionation, Sephadex G-200 gel filtration, DEAE-Sephadex A-50 ion exchange chromatography, and Hydroxyapatite adsorption chromatography. The purifying procedures resulted in 347-fold purification with the overall yield of 39.5% The purified enzyme had a specific activity(using ONPG as a substrate) of about 1, 585 units per mg protein. The molecular weight of the enzyme protein was estimated to be 140, 000 by gel filtration on Sephadex G-200, and SDS-polyacrylamide gel electorphoresis showed that the enzyme consisted of two identical subunits with a molecular weight of 72, 000.

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Purification of Therapeutic Serums of Snake Anti-Venom with Caprylic Acid

  • Norouznejad, Nilofar;Zolfagharian, Hossein;Babaie, Mahdi;Ghobeh, Maryam
    • Journal of Pharmacopuncture
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    • v.25 no.2
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    • pp.114-120
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    • 2022
  • Objectives: Antivenom serums have been used extensively for over a century and are the only effective treatment option for snake bites and other dangerous animal envenomations. In therapeutic serum centers, a wide range of antivenoms is made from animal serum, mainly equine and sheep, that are immunized with single or multiple venoms. This work aimed to use caprylic acid (CA) to purify therapeutic snake antivenom. Methods: Plasma was obtained from equine immunized with a mixture of venoms. Immunized plasma was obtained by precipitation of different concentrations (2-5%) of CA. This methodology was compared to that based on ammonium sulfate (AS) precipitation. Sediment plasma proteins were purified by ion-exchange chromatography. Protein assay, SDSPAGE, and agar gel diffusion were performed. Results: The total protein precipitation with AS was higher than precipitation with CA, but the best results were obtained when CA was added to the plasma until a final CA concentration of 5% was reached. Chromatography and electrophoresis indicated a stronger band for the 5% CA, and the gel diffusion assay showed antigen-antibody interaction in the purified serum. Conclusion: The use of CA compared to the routine method for purifying hyperimmune serums is a practical and cost-effective method for preparing and producing therapeutic serums. It constitutes a potentially valuable technology for alleviating the critical shortage of antivenom in Iran.

Study on removal method of Brownish black and White crust on Mural in Koguryo Tomb (고구려 고분벽화 오염물질 제거에 관한 연구)

  • Han, Kyeong-Soon;Lim, Kwon-Woong
    • Journal of Conservation Science
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    • v.22
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    • pp.99-108
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    • 2008
  • This research aimed to provide a scientific methodology for removing white and black/brown coloured stains on the wall paintings of tombs of Jinpari No 1 and No 4. in the Democratic People's Republic of Korea. For the analysis of chemical composition of stains of the samples from the wall paintings, a microscope and SEM/EDX were used. The analysis confirmed that the fomula of white coloured stains should be $CaSO_4$ or $CaCO_3$ and the black/brown coloured stains should be $CaSO_4$ or $CaCO_3$ with soil deposition. Because of the difficulties of testing several cleaning solutions on sample patches of large area of the painting, the author considered a risk-free cleaning solution as being the most appropriate one, with Ammonium bicarbonate and Anion exchange resin showing satisfactory cleaning effect without visible side effects. For the removal of dense layer of stains, the research suggested that physical cleaning should be followed by applying a cleaning solution.

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Removal of Ammonia from Aqueous Solutions with Zeolite and Bentonite (제오라이트 및 벤토나이트에 의한 수용액중 암모니아의 제거)

  • 이화영;오종기;김성규;고현백
    • Resources Recycling
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    • v.11 no.3
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    • pp.3-9
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    • 2002
  • Relnoval of ammonia from aqueous solutions has been studied with zeolite and bentonite minerals. Zeolite and bentonite powder were supplied by a domestic company and used as delivered without further purification. The aqueous pH was found to increase by addition of zeolite or bentonite up to pH 8.5 from initial pH of 5.5∼5.7. From the C.E.C. measurement by ammonium acetate leaching method, the values of C.E.C. of zeolite and bentonite sample were observed to be 129.7 meq/100 gr and 65.1 meq/100 gr, respectively and Na+ ion accounted for the major part of total C.E.C. in both cases. In the removal of ammonia with zeolite and bentonite, physical adsorption of ammonium ion onto minerals was believed to contribute to the removal of it as well as the intrinsic cation exchange reaction. Finally, zeolite was found to be superior to bentonite in the removal of ammonia from aqueous solutions.

Soil nitrogen availability in a thinned Larix leptolepis plantation using ion exchange resin bags (이온교환수지봉지를 이용한 일본잎갈나무 간벌지의 질소 유효도 측정)

  • Son, Yo-Whan;Kim, Hyun-Seop
    • Korean Journal of Environmental Agriculture
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    • v.19 no.2
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    • pp.188-190
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    • 2000
  • We explored changes in soil nitrogen (N) availability in a thinned (control, light, moderate, and heavy thinning) Larix leptolepis plantation determined by using ion exchange resin bags. Nitrogen availability varied among measurement periods, however, total available N (ammonium plus nitrate) concentrations did not change significantly in the 1 year since thinning. We found higher N availability in summer and fall than in winter.

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Study on desorption characteristics by mixed resins of active carbons and ion exchange resins for perchlorate ion (이온교환수지와 활성탄의 혼합수지를 이용한 과염소산 이온의 탈착 특성 연구)

  • Kim, Young-Eun;Jeong, Yu-Dong;Kim, Sun Hwan;Paeng, Ki-Jung
    • Analytical Science and Technology
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    • v.26 no.1
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    • pp.11-18
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    • 2013
  • Perchlorate ($ClO{_4}^-$) is the material that is used as propellants of rockets and material of explosive as a form of ammonium perchlorate salts. Ammonium perchlorate solution of high concentration is recovered from expired rocket through demilitarization process by the water-jet method. If people take perchlorate in food and water, it interferes with adsorption of iodide which is the substance needed to synthesize thyroid hormone in the thyroid gland. It has an bad influence upon disturbing pregnancy and synthesis of growth hormone. So the effective method is necessary to remove perchlorate anion in water. By considering economic aspect, we studied effective desorption (regeneration) of perchlorate anion from adsorbent with studies on removal and adsorption of perchlorate anion. Desorption experiment was conducted as batch type. Depending on various conditions (concentration, pH, cation anion form) elution, we evaluated amount, efficiency of desorption(amount of adsorption/desorption ${\times}$ 100). Also, research confirmed the efficiency of mixed resins between anion exchange resin and activated carbon and expected synergic effect from advantages of both adsorbents.