• Title/Summary/Keyword: PB-22

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Cloning, High-Level Expression, Purification, and Properties of a Novel Endo-${\beta}$-1,4-Mannanase from Bacillus subtilis G1 in Pichia pastoris

  • Vu, Thi Thu Hang;Quyen, Dinh Thi;Dao, Thi Tuyet;Nguyen, Sy Le Thanh
    • Journal of Microbiology and Biotechnology
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    • v.22 no.3
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    • pp.331-338
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    • 2012
  • A novel gene coding for an endo-${\beta}$-1,4-mannanase (manA) from Bacillus subtilis strain G1 was cloned and overexpressed in P. pastoris GS115, and the enzyme was purified and characterized. The manA gene consisted of an open reading frame of 1,092 nucleotides, encoding a 364-aa protein, with a predicted molecular mass of 41 kDa. The ${\beta}$-mannanase showed an identity of 90.2-92.9% ${\leq}95%$) with the corresponding amino acid sequences from B. subtilis strains deposited in GenBank. The purified ${\beta}$-mannanase was a monomeric protein on SDS-PAGE with a specific activity of 2,718 U/mg and identified by MALDI-TOF mass spectrometry. The recombinant ${\beta}$-mannanase had an optimum temperature of $45^{\circ}C$ and optimum pH of 6.5. The enzyme was stable at temperatures up to $50^{\circ}C$ (for 8 h) and in the pH range of 5-9. EDTA and most tested metal ions showed a slightly to an obviously inhibitory effect on enzyme activity, whereas metal ions ($Hg^{2+}$, $Pb^{2+}$, and $Co^{2+}$) substantially inhibited the recombinant ${\beta}$-mannanase. The chemical additives including detergents (Triton X-100, Tween 20, and SDS) and organic solvents (methanol, ethanol, n-butanol, and acetone) decreased the enzyme activity, and especially no enzyme activity was observed by addition of SDS at the concentrations of 0.25-1.0% (w/v) or n-butanol at the concentrations of 20-30% (v/v). These results suggested that the ${\beta}$-mannanase expressed in P. pastoris could potentially be used as an additive in the feed for monogastric animals.

Use of hybrid materials in the trace determination of As(V) from aqueous solutions: An electrochemical study

  • Tiwari, Diwakar;Jamsheera, A.;Zirlianngura, Zirlianngura;Lee, Seung Mok
    • Environmental Engineering Research
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    • v.22 no.2
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    • pp.186-192
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    • 2017
  • The carbon paste electrode (CPE) was modified with the pristine bentonite and hybrid material (HDTMA-modified bentonite). The modified-CPEs are then employed as working electrode in an electrochemical detection of As(V) from aqueous solutions using the cyclic voltammetric measurements. Cyclic voltammograms revealed that As(V) showed reversible behavior onto the working electrode. The hybrid material-modified carbon paste electrode showed significantly enhanced electrochemical signal which was then utilized in the low level detection of As(V). Moreover, the studies were conducted at neutral pH conditions. The electrochemical studies were conducted with scan rates (20 to 200 mV/s) to deduce the mechanism of redox processes involved at the electrode surface. The anodic current was linearly increased, increasing the concentration of As(V) from 5.0 to $35.0{\mu}g/g$ using the hybrid material-modified electrode. This provided fairly a good calibration line for As(V) detection. The presence of varied concentrations of As(III) in the determination of total arsenic was studied. The influence of several cations and anions viz., Cu(II), Mn(II), Zn(II), Pb(II), Cd(II), Fe(III), $Cl^-$, $NO_3{^-}$, $PO_4{^{3-}}$, EDTA and glycine in the detection of As(V) from aqueous solution was also studied. Further, in an attempt to simulate the real matrix analysis, the tap water sample was spiked with As(V) and subjected for As(V) detection using the modified-CPE.

STUDIES ON THE WATER QUALITY OF NAGDONG RIVER DOWNSTREAM FOR DRINKING WATER AND INDUSTRIAL SUPPLY WATER 2. ANNUAL VARIATIONS OF HEAVY METAL CONCENTRATIONS IN THE DOWNSTREAM WATER OF NAGDONG RIVER FROM MAY 1977 TO APRIL 1978 (음료수및 공업용수로서의 낙동강 하류수질에 대하여 2. 남지이남 낙동강 하류수의 중금속 함량에 대하여 (1977년 5월~1978년 4월))

  • WON Jong Hun;YANG Han Seob
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.11 no.3
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    • pp.139-146
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    • 1978
  • The annual variations of some heavy metal concentrations, Al, Mn, Fe, Cu, Pb, Zn, Cd, Hg were determined on tile day of spring tides in every month from May 1977 to April 1978 at eight stations in the Nagdong River downstream. Samples were taken at intervals of one or two hours from 7 a.m. to 7 p.m. at each station. Annual ranges and means of concentrations of the heavy metals are as follows : aluminium 40.7-3700 ppb, 286 ppb; manganese 10.3-261.5 ppb, 80.8 ppb; iron ND-1237 ppb, 147.7 ppb; copper ND-30.9 ppb, 2.49 ppb; lead ND-29.9 ppb, 1.10 ppb; zinc ND-156.8 ppb, 5.61 ppb; cadmium ND-1.22 ppb, 0.09 ppb; mercury ND-0.37 ppb, 0.02 ppb respectively. In general, the contents of heavy metals except managanese and mercury were higher at the stations above station one, Kupo, though the ranges of the contents showed remarkable difference according to the sampling stations. Annual means of the concentrations of iron and manganese were exceeded already the desirable standards for industrial water and closed to the criteria of raw water for public supply. The values that over the criteria of raw water were sometimes found. Mercury concentration showed comparative high level such as ND-0.37 ppb in annual range but it was not over the environment criterion, 0.5 ppb. Aluminium, iron and mercury showed generally high values in summer period and other metals in winter period, through the patterns of seasonal variations were difference depending on the sampling stations and the kind of metals.

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Experimental Study on Four Cation Exchange Membranes in Electrosynthesis of Ammonium Persulfate

  • Wang, Chao;Zhou, Junbo;Gao, Liping
    • Journal of Electrochemical Science and Technology
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    • v.9 no.1
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    • pp.37-43
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    • 2018
  • In order to improve current efficiency and decrease energy consumption in the electrosynthesis of ammonium persulfate, electrolytic properties of four cation exchange membranes, namely, the $JCM-II^{(R)}$ membrane, $Nafion^{(R)}$ 324 membrane CMI-$7000^{(R)}$ membrane and a self-made perfluorosulfonic ion exchange membrane (PGN membrane) were investigated using a sintered platinized titanium anode and a Pb-Sb-Sn alloy cathode in a self-made electrolytic cell. The effect of cell voltage and electrolyte flow rate on the current efficiency and the energy consumption were investigated. The results indicated that the PGN membrane could improve current efficiency to 94.85% and decrease energy consumption to $1119kWh\;t^{-1}$ (energy consumption per ton of the ammonium persulfate generated) under the optimal operating conditions and the highest current efficiency of the $JCM-II^{(R)}$ membrane, $Nafion^{(R)}$ 324 membrane and CMI-$7000^{(R)}$ membrane were 80.73%, 77.76% and 73.22% with their lowest energy consumption of $1323kWh\;t^{-1}$, $1539kWh\;t^{-1}$ and $2256kWh\;t^{-1}$, respectively. The PGN membrane has the advantages of high current efficiency and energy power consumption and has sufficient mechanical strength with the reinforced mesh. Therefore the PGN membrane will has good value in popularization in the industrial electrosynthesis of ammonium persulfate in the future.

The Concentrations of TSp, PM10 and Heavy Metal at Underground parking Lots of Public Facilities in Taegu City (대구시 공중이용시설 지하주차장의 총부유먼지, 호흡성먼지 및 중금속 농도)

  • 이현주;정재열;이종영;송희봉;홍성철
    • Journal of Environmental Health Sciences
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    • v.26 no.2
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    • pp.22-29
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    • 2000
  • This study was performed to investigate the concentrations of TSP, PM10 and heavy metals(Pb, Cd,Hg, Ni, Zn) of underground parking lots in Taegu city. The samples were collected from 3 department stores and 1 central park in the winter and the summer, 1997. The samples of 3 department stores were divided into sale period and non-sale period in the winter and the summer. The concentrations of TSP and PM10 were 109.6±1.5㎍/㎥ and 93.3±1.5㎍/㎥. In TSP, the zinc was the highest level, 287.16±1.5ng/㎥ and the cadmium was the lowest, 2.4±2.1ng/㎥ and in PM10, the zinc was the highest level, 193.6±1.5ng/㎥ and the cadmium was the lowest, 0.6±3.9ng/㎥. The correlation coefficient between the concentration of TSP and PM10 was 0.982(p<0.05). The correlation coefficients of corresponding heavy metal in TSP and PM10 were 0.863 for lead, 0.617 for mercury, 0.890 for nickel and 0.850 for zinc(p<0.05). The concentrations of TSP, PM10 and cadmium of PM10 in the winter were higher than those of the summer. However, the concentration of Hg of PM10 in the summer was higher than that of the winter. The concentrations of TSP and PM10 in sale period was higher than those of non-sale period and the concentrations of heavy metal in TSP and PM10 were not significantly different between sale and non-sale period. The proportions of PM10 to TSP were above 60% in dust, lead, nickel, and zinc and less than 40% in cadmium and mercury. The concentrations of TSP, PM10 and heavy metal in the underground parking lots were comparatively lower than those of general atmosphere. However, this kind of research to PM10 in the underground parking lots must be continued because it is very important particulate matter that affects human beng's health.

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Improved Enucleation Efficiency of Pig Somatic Cell Nuclear Transfer by Early Denudation of Oocytes at 30 Hours of In Vitro Maturation

  • Song, Kil-Young;Hyun, Sang-Hwan;Lee, Eun-Song
    • Journal of Embryo Transfer
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    • v.22 no.4
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    • pp.235-243
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    • 2007
  • Our goal was to examine the effects of early denudation on the enucleation efficiency and developmental competence of embryos following somatic cell nuclear transfer (SCNT) and parthenogenetic activation (PA). Oocytes were denuded following 30 h of in vitro maturation (IVM) and then cultured with (D+) or without (D-) their detached cumulus cells for additional $10{\sim}14$ h. Control oocytes were denuded after $40{\sim}44$ h of IVM. The size of the perivitelline space was larger at 40 h of IVM ($11.7{\sim}11.8{\mu}m$) than at 30 h ($8.9{\mu}m;$ p<0.01). The distances between the metaphase II (M II) plates and the polar bodies (PBs) were shorter in D+ ($19.4{\mu}m$) and D- oocytes ($18.9{\mu}m$) than in control oocytes ($25.5{\mu}m;$ p<0.01). Enucleation rates following blind aspiration at 40 h of IVM were higher (p<0.01) in D+ (92%) and D- oocytes (93%) compared to controls (82%). Early denudation did not affect oocyte maturation or the in vitro development of SCNT and PA embryos. When SCNT embryos from D+ oocytes were transferred to four gilts, pregnancy was established in two pigs, and one of them farrowed three live piglets. In conclusion, early denudation of oocytes at 30 h of IVM could improve the enucleation efficiency by maintaining the M II plate and the PB within close proximity and support the in vivo development of SCNT embryos to term.

Characterization of a Recombinant Thermostable Xylanase from Hot Spring Thermophilic Geobacillus sp. TC-W7

  • Liu, Bin;Zhang, Ningning;Zhao, Chao;Lin, Baixue;Xie, Lianhui;Huang, Yifan
    • Journal of Microbiology and Biotechnology
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    • v.22 no.10
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    • pp.1388-1394
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    • 2012
  • A xylanase-producing thermophilic strain, Geobacillus sp. TC-W7, was isolated from a hot spring in Yongtai (Fuzhou, China). Subsequently, the xylanase gene that encoded 407 amino acids was cloned and expressed. The recombinant xylanase was purified by GST affinity chromatography and exhibited maximum activity at $75^{\circ}C$ and a pH of 8.2. The enzyme was active up to $95^{\circ}C$ and showed activity over a wide pH range of 5.2 to 10.2. Additionally, the recombinant xylanase showed high thermostability and pH stability. More than 85% of the enzyme's activity was retained after incubation at $70^{\circ}C$ for 90 min at a pH of 8.2. The activity of the recombinant xylanase was enhanced by treatment with 10 mM enzyme inhibitors (DDT, Tween-20, 2-Me, or TritonX-100) and was inhibited by EDTA or PMSF. Its functionality was stable in the presence of $Li^+$, $Na^+$, and $K^+$, but inhibited by $Hg^{2+}$, $Ni^{2+}$, $Co^{2+}$, $Cu^{2+}$, $Zn^{2+}$, $Pb^{2+}$, $Fe^{3+}$, and $Al^{3+}$. The functionality of the crude xylanase had similar properties to the recombinant xylanase except for when it was treated with $Al^{2+}$ or $Fe^{2+}$. The enzyme might be a promising candidate for various industrial applications such as the biofuel, food, and paper and pulp industries.

The Microstructure and Interfacial Reaction between Sn-3.5wt.%Ag-1wt.%Zn and Cu Substrate (Sn-3.5wt.%Ag-1wt.%Zn 땜납과 Cu기판간의 미세조직 및 계면반응)

  • Baek, Dae-Hwa;Seo, Youn-Jong;Lee, Kyung-Ku;Lee, Doh-Jae
    • Journal of Korea Foundry Society
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    • v.22 no.2
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    • pp.89-96
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    • 2002
  • This study examined the effects of adding Zn to Sn-3.5Ag solder on the microstructure changes and behavior of interface reaction of the solder joint with Cu substrate. The solder/Cu joints were examined with microscope to observe the characteristics of microstructure changes and interfacial reaction layer with aging treatment for up to 120 days at $150^{\circ}C$. Results of the microstructure changes showed that the microstructures were coarsened with aging treatment, while adding 1%Zn suppresses coarsening microstructures. The Sn-3.5Ag/Cu had a fast growth rate of the reaction layer in comparison with the Sn-3.5Ag-1Zn at the aging temperature of $150^{\circ}C$. Through the SEM/EDS analysis of solder joint, it was proved that intermetallic layer was $Cu_6Sn_5$ phase and aged specimens showed that intermetallic layer grew in proportion to $t^{1/2}$, and the precipitate of $Ag_3Sn$ occur to both inner layer and interface of layer and solder. In case of Zn-containing composite solder, $Cu_6Sn_5$ phase formed at the side of substrate and Cu-Zn-Sn phase formed at the other side in double layer. It seems that Cu-Zn-Sn phase formed at solder side did a roll of banrier to suppress the growth of the $Cu_6Sn_5$ layer during the aging treatment.

Human Multi-route Exposure Assessment of Lead and Cadmium for Korean Volunteers (일부 자원자를 통한 납, 카드뮴의 다경로 인체 노출평가)

  • Oh, Eun-Ha;Lim, Ho-Sub;Jang, Jae-Yeon;Lee, Eun-Il
    • Journal of Preventive Medicine and Public Health
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    • v.39 no.1
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    • pp.53-58
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    • 2006
  • Objectives: We wanted to evaluate the main route of exposure to lead and cadmium for the general population in Korea by using multi-route and multi-media exposure assessment Methods: Samples of air at the homes, samples of the food and water and peripheral blood samples were collected from thirty volunteers living in Seoul (the metropolitan area), Yong-in (the suburban area) and Ansan (the industrial area) in 2001. Graphite furnace atomic absorption spectrometric methods were used for the determination of the Pb and Cd levels in the air, food, water and blood samples. Results: The average intake of lead through the air, drinking water and food were $5.06{\mu}g/day\;(26.3%)\;0.002{\mu}g/day\;(0.1%),\;and\;16.4{\mu}g/day\;(73.6%)$, respectively. The average intake of cadmium through the air, drinking water and food were $0.082{\mu}g/day\;(0.9%),\;0.001{\mu}g/day\;(0.007%),\;and\;12.61 {\mu}g/day\;(99.0%)$, respectively. The blood lead level was statistically higher in the male subjects than in the female subjects (3.39 and $2.22{\mu}g/dl$, respectively), and only gender was a significant variable on the multiple regression analysis for blood lead. Conclusions: This study showed that the intake of lead and cadmium through food was the major route of exposure. A policy to reduce the pollutants according to the exposure routes should be established. However, more studies will be needed to support these data for the general population.

Effects of Onion Juice on Toxicity of Lead in Rat (양파즙 투여가 Rat의 납 독성에 미치는 영향)

  • 서화중;임현지;정두례
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.22 no.2
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    • pp.138-143
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    • 1993
  • In this study the onion juice (2%) in diet fed rats simultaneously ingested lead acetate 100mg/ kg (OP group) showed more increased weight gain than single lead treated rats (P group). The OP group had also improved in the hemoglobin contents and biochemical analyzed values of blood including GPT, blood urea nitrogen and alkaline phosphatase, which were elevated in case of P group rats. The Pb content in the rats liver of OP group was lower than in the rats liver of P group. In the histopathological findings of liver cell OP group rats did not show any signs of liver damage as observed in P group rats that had degenerated hepatocytes, followed sinusoidal dilatation, perivascular hemorrhage and some necrosis of hepatic cells accompanied by increased Kuffer cell bearing dark brown pigment. In conclusion 2% onion juice diet in rat have somewhat antidotic effects on the lead intoxicated rats.

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