• Title/Summary/Keyword: Pb band

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Separation and Enzymological Characteristics of Polygalacturonase by Aspergillus sp. (Aspergillus속이 생산하는 Polygalacturonase의 분리 및 특성)

  • 차원섭;김진구;박준희;오상룡;천성숙;조영제
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.24 no.4
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    • pp.570-577
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    • 1995
  • Aspergillus sp. SB-2704 was selected for its strong polygalacturonase activity among various strain of mold found in soil. It was found that production of polygalacturonase reached to maximum when the wheat bran medium containing 1% polypepton, 1% glucose, and 0.2% FeSO4 were cultured for 3 days at 35$^{\circ}C$. Polygalacturonase was purified 20.90 fold from Aspergillus SB-2704. The purification procedures include ammonium sulfate treatment, gel filtration on Sephdex G-150 and DEAE-cellulose ion exchange chromatography. Yield of the enzyme purification was 4.34%. Purified enzyme was confirmed as a single band by the polyacrylamide gel electrophoresis. When the purified enzyme was applied to SDS-polyacrylamide gel electrophoresis, the molecular weight was estimated to be 36,000. The optimum pH for the enzyme activity was 5.5 and optimum temperature was 5$0^{\circ}C$. The enzyme is stable in acidic condition. The activity of purified enzyme was inhibited by Pb2+, Hg2+ and Ba2+, whereas activated by Cu2+, Mn2+, Mg2+ and Fe2+. The activity of polygalacturonase was inhibited by the treament wit maleic anhydride, iodine, and EDTA. The result indicate the possible involvement of histidine and metal ion at active site.

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Optical Properties of Metal Halide Perovskite Nanocrystals with Addition of Metal Bromide (금속 브롬화물의 첨가에 따른 금속 할라이드 페로브스카이트 나노결정의 광학적 특성 변화)

  • Yun, Seokjin;Choi, Jihoon
    • Journal of the Korean institute of surface engineering
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    • v.52 no.6
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    • pp.334-341
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    • 2019
  • Organometal halide perovskite materials have attracted much attention in the photovoltaic and light emitting devices due to the compositional flexibility with AMX3 formula (A is an organic amine cation; M is a metal ion; X is a halogen atom). The addition of homovalent or heterovalent metal cations to the bulk organohalide perovskites has been performed to modify their energy band structure and the relevant optoelectronic properties by ligand-assisted ball milling. Here, we report CH3NH3Pb1-xMxBr3 nanocrystals substituted by metallic cations (M is Sn2+, In3+, Bi3+; x = 0, 0.01, 0.02, 0.05, 0.1, 0.2). Photoluminescence and quantum yield was significantly reduced with increasing metallic cations content. These quenching effect could be resulted from the metal cations that behave as a non-radiative recombination center.

Chemical Composition of Cultured and Wild Codonopsis lanceolata Roots of Different Age Groups -I. Proximate Composition, Minerals and Protein Fractions- (더덕(沙蔘)의 년근별(年根別) 화학성분(化學成分)에 관(關)한 연구(硏究) -제1보(第1報) : 일반성분(一般成分), 무기질(無機質) 및 단백질(蛋白質) 분획(分劃)-)

  • Park, Boo-Duck;Park, Yong-Gone;Choi, Kwang-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.14 no.3
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    • pp.274-279
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    • 1985
  • Proximate compositions, minerals and protein fractions of the roots of cultured and wild Codonopsis lanceolata of different age groups were examined as the basic research for the study of their source of processed foods. The most abundant proximate composition of the roots of C. lanceolata was observed to be total sugars and next come crude protein, crude fiber, crude fat and ash in descending order irrespective of cultured and wild ones. The richest mineral contained in the roots was noticed to be K and followed by Mg and Ca. Generally increased tendency of crude protein, fat, ash, K, Mg, Ca, Mn, Zn, Cu and P contents were observed with older roots, however, decreased total sugars and Fe content. Lead and cadmium content was far bellow the authorized tolerance limits. The quantitative fractionation of the protein of the roots ranked albumin the highest content, followed by globuin, prolamin and glutelin. Decreased albumin content was observed with the older age roots, while increased globulin, prolamin and glutelin content. The minimum solubility of the soluble protein of the roots was found to be at pH 4.0 and maximum, at pH 10.0. Disc gel electrophoresis of the soluble protein of C. lanceolata roots showed almost similar patterns and numbers of bands. The molecular weight for main band protein was estimated to be about 90,000.

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Deterimination of an Optimal Time Point for Analyzing Transcriptional Activity and Analysis of Transcripts of Avian Influenza Virus H9N2 in Cultured Cell (배양세포에서 Semi-quantitative RT-PCR에 의한 조류인플루엔자 H9N2의 전사활성 분석 최적 시기 결정 및 전사체 분석)

  • Na, Gi-Youn;Lee, Young-Min;Byun, Sung-June;Jeon, Ik-Soo;Park, Jong-Hyeon;Cho, In-Soo;Joo, Yi-Seok;Lee, Yun-Jung;Kwon, Jun-Hun;Koo, Yong-Bum
    • Korean Journal of Microbiology
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    • v.45 no.3
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    • pp.286-290
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    • 2009
  • The transcription of mRNA of avian influenza virus is regulated temporally during infection. Therefore, the measurement of transcript level in host cells should be performed before viral release from host cells because errors can occur in the analysis of the transcript levels if the viruses released from the infected cells re-infect cells. In this study, the timing of viral release was determined by measuring the level of viral RNA from viruses released from H9N2-infected chicken fibroblast cell line UMNSAH/DF-1 by semi-quantitative RT-PCR. The viral genomic RNA was isolated together with mouse total RNA which was added to the collected medium as carrier to monitor the viral RNA recovery and to use its GAPDH as an internal control for normalizing reverse transcription reaction as well as PCR reaction. It was found that viral release of H9N2 in the chicken fibroblast cell line UMNSAH/DF-1 took between 16 and 20 h after infection. We measured all 8 viral mRNA levels. Of the 8 transcripts, 7 species of viral mRNAs (each encoding HA, NA, PB1, PB2, NP, M, NS, respectively) except PA mRNA showed robust amplification, indicating these mRNA can be used as targets for amplification to measure transcript levels. These results altogether suggest that the method in this study can be used for screening antiviral materials against viral RNA polymerase as a therapeutic target.

Characteristics of Microbial Pretense far Application to Abolished Protein Resource (폐단백자원에 이용하기 위한 미생물 Protease의 특성)

  • Chun, Sung-Sook;Cho, Young-Je;Sung, Tae-Soo;Son, Jun-Ho;Choi, Cheong
    • Applied Biological Chemistry
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    • v.41 no.1
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    • pp.6-12
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    • 1998
  • To extract insoluble proteins and to improve functional properties of abolished proteins, a protease producing Aspergillus sp. MS-18 was isolated from soil. The enzyme was purified and its enzymological characteristics were investigated. It was found that production of protease reached to the maximum when the wheat brae medium containing, 3% arabinose, 0.5% polypepton, 0.1% $(NH_4)_2SO_4$ and 0.2% magnesium chloride was cultured for 3 days. Protease was purified 16.9 folds after ion exchange chromatography and gel filtration and the specific activity was 340.4 unit/mg. Purified enzyme was confirmed as a single band by the polyacrylamide gel electrophoresis. The molecular weight of protease was estimated to be 30,000. Crystalization form of purified protease was a stick shape with rounding edges. The optimum pH and temperature for the protease activity were 9.0 and $60^{\circ}C$, respectively. The enzyme was stable in pH 7.0-12.0 at $50^{\circ}C$. The activity of purified enzyme was inhibited by $Hg^{2+}$, $Cu^{2+}$, $Zn^{2+}$ and $Pb^{2+}$, whereas it was activited by $Na^+$, $Mg^{2+}$ and $Mn^{2+}$. The activity of the protease was inhibited by the treatment with ethylenediaminetetraacetic acid and phenylmethane sulfonyl fluoride. The result suggests that the purified enzyme is a serine protease with metal ion at active site. Km and Vmax of purified protease were $29.33\;{\mu}mole/L$ and $5.13\;{\mu}g/min$, respectively.

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Effects of Heavy Metals on Mortality and Metabolite Changes in Mosquitoes, Culex pipiens pallens (Diptera: Culicidae) (중금속이 빨간집모기(Culex pipiens)의 치사율과 대사물질의 변화에 미치는 영향)

  • Lee, Su Mi;Shin, Byung Sik
    • Korean Journal of Environmental Biology
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    • v.36 no.4
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    • pp.694-700
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    • 2018
  • To investigate the effect of heavy metals (Cu, Cd, Hg, Pb, Zn) on mosquito Culex pipiens, 50% lethal concentration ($LC_{50}$) concentration, total lipid content, lipid content composition, and total protein content were measured. The results showed that the Hg $LC_{50}$ was $0.45mg\;kg^{-1}$ and the Hg toxicity was higher than the other metals. The results also showed that cadmium (Cd) significantly retarded the growth of mosquito larvae among the tested heavy metals. Six types of lipid bands were isolated from mosquito samples exposed to heavy metals, and five of them were identified (phospholipid, cholesterol, fatty acid, triglyceride, and cholesterol ester). The total lipid content of all treatments decreased compared to the control. In addition, the protein content of the control group ($0.51mg\;ind.^{-1}$) was higher than that of the treated group, and the protein content of the larvae treated with Hg and Cd was very low. In conclusion, heavy metals not only lower the survival rate of mosquitoes, but also affect the protein content and lipids in the mosquito's body, thereby causing growth inhibition. These results indicate that heavy metal contamination will affect the size of mosquito population by inhibiting the survival and growth of mosquitoes, and it is expected that it can be used to measure the degree of contamination using mosquito through subsequent studies.

Purification and Characteristics of Endo-Polygalacturonase from Korean Tomato (한국산 토마토의 Endo-Polygalacturonase 정제 및 성질)

  • Choi, Cheong;Cho, Young-Je;Son, Gyu-Mok
    • Applied Biological Chemistry
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    • v.33 no.1
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    • pp.73-78
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    • 1990
  • Endo-polygalacturonase was purified from tomato, Lycopersicon esculentum L. The purification procedures included gel filtration on Sephadex G-150 and DEAE-cellulose ion exchange chromatography. Yield of the enzyme purification was 12.74 %. Purified enzyme was confirmed as a active single band by the SDS-polyacrylamide gel electrophoresis. When the purified enzyme was applied to SDS-PAGE, the molecular weight was estimated about 50,000. The optimum pH for the enzyme activity was 5.0 and the range of its stability to the pH was 4.0 to 5.0. The optimum temperature was $50^{\circ}C$, while the enzyme was abruptly inactivated above $50^{\circ}C$. From the result of the study on the effects of metals ion, it was found that $Ag^+$, $Zn^{++}$ and $Mg^{++}$ increased on the enzyme activity. In contrast, $Ba^{++}$, $Hg^{++}$, $Pb^{++}$, $Ca^{++}$, $Mn{++}$, $Cu^{++}$, $Fe^{+++}$, $Na^+$ and $K^+$ decreased it. the reaction catalyzed by this enzyme followed typical Michaelis-Menten kinetics with the Km value of $1.43{\times}10^{-1}\;mol/l$.

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Studies on proteolytic enzyme from A New Strain in Serratia sp. (Serratia속의 신균주가 생산하는 단백질분해효소)

  • 최완수;정계종;이주경;박주웅;이상훈;이진복;이송락;최신원
    • YAKHAK HOEJI
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    • v.37 no.2
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    • pp.129-135
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    • 1993
  • Serratia sp. Y-4 was isolated from soil. Many characteristics of the strain and optimum cultivation condition for protease production were investigated.,The protease from Serratia sp. Y-4 was purified and studied for the properties of the enzyme. The isolated strain was identified to the genus Serratia. The strain was cultivated in 1%-casein, 0.5%-Na$_{3}$PO$_{4}$.7H$_{2}$O, 0.1%-NaCl, 0.05%-KCI, 0.02%-MgSO$_{4}$.7H$_{2}$O, 0.02%-CaCl$_{2}$.2H$_{2}$O, 0.02%-ZnSO$_{4}$.7H$_{2}$O, 0.02%-MnCl$_{2}$.4H$_{2}$O and 0.5%-soy bean oil at pH 7.0 for 35 hrs. The enzyme was purified about 5.89 fold from the culture broth with 31.1% recovery and 19,613 u/mg through ultrafiltration, ammonium sulfate, DEAE-sephacel and Superose-12 chromatography. The purified enzyme was identified as one band by isoelectric focusing, SDS- and native-PAGE. It has maxium activity at $37^{\circ}C$ and pH 9.0. Molecular weight of it is approx. 50 kD and pl is about 6.70. Its Km value for casein was 20 mg/ml. 5 mM-EDTA, 5mM-SDS, Ag$^{+1}$, Cu$^{+2}$, Hg$^{+2}$ and Pb$^{+2}$ inhibited the enzyme.

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Purification and Characterization of Transglutaminase from a Newly Isolated Streptomyces platensis YK-2 (토양 방선균 Streptomyces platensis YK-2가 생산하는 Transglutaminase의 정제 및 효소학적 특성)

  • Ko, Hee-Sun;Kim, Hyun-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.6
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    • pp.801-806
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    • 2009
  • A species producing transglutaminase (EC 2.3.2.13) was isolated from forest soil and identified as Streptomyces platensis YK-2. The transglutaminase was purified from culture broth by 50% methanol precipitation, followed by successive chromatography on DEAE-Sephadex. The yield and purification-fold was 63.4% and 2.2-fold, respectively. The purified microbial transglutaminase (MTG) migrated as a single band of approximately 45 kDa upon sodium dodecyl sulfate polyacrylamide gel eletrophoresis. The isoelectric point determined by multichambered electrofocusing was pH $6.0{\sim}7.0$. The enzyme was strongly inhibited by $Hg^{++}$, but was activated by $Cd^{++}$, $Mg^{++}$, $Mn^{++}$, $Pb^{++}$ and reducing agents such as dithiothreitol and mercaptoethanol.

Uptake of Heavy Metal Ions by Water Dropwort (Oenanthe stolonifera DC.) and Identification of Its Heavy Metal-Binding Protein (미나리의 중금속 흡수량 측정 및 중금속 결합단백질의 동정)

  • Park, Young-Il;Kim, Hee-Guen;Kim, Yoo-Young;Kim, In-Soo
    • Applied Biological Chemistry
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    • v.39 no.6
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    • pp.494-500
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    • 1996
  • Uptake of hen metal ions by water dropwort (Oenanthe stolonifera DC.) and its cadmium-binding protein were studied to probe for good method to remove heavy metal contaminants from environments. The plant was cultured in the culture medium (pH 7.0) containing the various concentrations of $Cd^{2+}$, $Cr^{3+}$ or $Pb^{2+}$, for 3 and 7 days. The residual heavy metals deposited in roots linearly increased as the metal ions concentration increased up to 17 ppm for $Cd^{2+}$, 20 ppm for $Cr^{3+}$ and 50 ppm for $Pb^{2+}$. Above these concentrations, the plant growth was inhibited and the uptake rates of the metal ions decreased. The heavy metals absorbed by the plant were mostly deposited in roots. In particular, the residual concentration of lead in roots was about four times higher than those of cadmium and chromium. When cultured in the medium containing 20 ppm of each metal ion, 80% of cadmium, 90% of cromium and 96% of lead were deposited in roots out of the total residual metal ions in the plant. These values correspond to 6.1 mg of cadmium, 5.2 mg of chromium and 23.6 mg of lead per one gram of roots tissue on a dry weight basis. A cadmium-binding protein was partially purified by extraction, gel filtration and DEAE-Cellulose chromatography from water dropworts that was grown in the medium containing 20 ppm $Cd^{2+}$. The purified protein was a single band on SDS- and non-denaturing- polyacrylamide gel electrophoresis. Its molecular mass was estimated to be ca. 5,000 dalton by gel filteration. Analysis of amino acid composition of the protein indicated that it had a typical amino acid composition of heavy metal-binding protein in that it contained 27% of acidic amino acids and 9.9% of cysteine. However, it is likely that the protein is a new plant metal-binding protein, since its amino acid composition is somewhat different from those of phytochelatins that have been known so far.

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