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A Novel cry2Ab Gene from the Indigenous Isolate Bacillus thuringiensis subsp. kurstaki

  • Sevim, Ali;Eryuzlu, Emine;Demirbag, Zihni;Demir, Ismail
    • Journal of Microbiology and Biotechnology
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    • v.22 no.1
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    • pp.133-140
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    • 2012
  • A novel cry2Ab gene was cloned and sequenced from the indigenous isolate of Bacillus thuringiensis subsp. kurstaki. This gene was designated as cry2Ab25 and its sequence revealed an open reading frame of 1,902 bp encoding a 633 aa protein with calculated molecular mass of 70 kDa and pI value of 8.98. The amino acid sequence of the Cry2Ab25 protein was compared with previously known Cry2Ab toxins, and the phylogenetic relationships among them were determined. The deduced amino acid sequence of the Cry2Ab25 protein showed 99% homology to the known Cry2Ab proteins, except for Cry2Ab10 and Cry2Ab12 with 97% homology, and a variation in one amino acid residue in comparison with all known Cry2Ab proteins. The cry2Ab25 gene was expressed in Escherichia coli BL21(DE3) cells. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed that the Cry2Ab25 protein is about 70 kDa. The toxin expressed in BL21(DE3) exhibited high toxicity against Malacosoma neustria and Rhagoletis cerasi with 73% and 75% mortality after 5 days of treatment, respectively.

Mechanical Properties of Carbon Fiber/Nylon 6 Composite Introducing Coupling Agent (II) -Increasing Interfacial Strength of Composite- (카플링제를 도입한 탄소섬유/나일론 6 복합재료의 기계적 성질(II) -복합재료의 계면강도 증가-)

  • Park, Chan Hun;Lee, Yang Hun;Shin, Eun Joo
    • Textile Coloration and Finishing
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    • v.9 no.4
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    • pp.47-53
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    • 1997
  • To improve the interfacial bonding of carbon fiber-nylon 6 composite, carbon fiber(CF) were oxidized by nitric acid treatment, and two types of graft polymer(GP) of nylon 6-g-polyacrylamide (PAAm) -water dispersable GP(WDGP) and m-cresol solu ble GP(CSGP) were treated as coupling agents. Introduction of polar groups such as -COOH, -OH, etc, on the surface of the oxidized CF was confirmed by IR spectra. The stem polymer of nylon 6 in the coupling agent (GP) could be compatible with'matrix nylon 5, and the grafted branch of PAAm on GP could react to the polar groups on the oxidized CF in composite. The interfacial strength was measured by the transverse tensile test to the fiber direction for single CF embedded nylon 6 film especially prepared and by the pull-out test method. The interfacial strength of the composite reinforced with oxidized CF is greater than that reinforced with unoxidized CF. The interfacial strength of the composite was increased by treatment of coupling agents(GPs) considerably, and the increasing tendency by the WDGP is greater than that by the CSGP. The optimum conditions of coupling agent treatment are as follows: the concentration, adsorption tlme of GP, and curing temperature are 2%, 20 minutes, and $170^{\circ}$, respectively.

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Stability of Antioxidant Properties and Essential Amino Acids in Squid Viscera Hydrolysate Produced using Subcritical Water

  • Kim, Ryoung-Hee;Asaduzzaman, A.K.M.;You, Chi-Hong;Chun, Byung-Soo
    • Fisheries and Aquatic Sciences
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    • v.16 no.2
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    • pp.71-78
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    • 2013
  • Subcritical water hydrolysis was carried out in a batch reactor to produce valuable materials, such as low-molecular weight (MW) peptides and essential amino acids with antioxidant properties, from heat-dried squid viscera. Hydrolysis of squid viscera was performed at 160 to $280^{\circ}C$ for 3 min. The yield was increased by increasing the temperature and pressure, while the protein content of squid viscera hydrolysate decreased with increasing temperature. Low-MW peptides were detected in all hydrolysates by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The highest yields of free and structural amino acids in heat-dried squid viscera hydrolysate were at $160^{\circ}C$ and were $411.95{\pm}1.15$ and $346.62{\pm}1.25$ mg/100 g, respectively. All essential amino acids were detected in viscera hydrolysates; leucine was the most abundant. Antioxidant activities of hydrolysates were highest at $220^{\circ}C$. Greater than $98{\pm}0.26%$ of the ABTS antioxidant activity was retained in hydrolysates after long-term heat treatment.

Pseudomonas sp. CB-33이 생산하는 $\beta$-Xylosidase의 특성

  • Yu, Jin-Whan;Kim, Hyun-Ku;Kim, Chi-Kyung;Lim, Jai-Yun
    • Microbiology and Biotechnology Letters
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    • v.24 no.2
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    • pp.197-205
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    • 1996
  • The $\beta$-xylosidase was purified 99- fold from the culture supernatant of Pseudo onas sp. CB-33 by ammonium sulfate precipitation, PEI precipita- tion, DEAE-Sephadex column chromatography, Sephadex G-75 gel filtration chromatography and preparative disc gel electrophoresis. Molecular weight of the enzyme was estimated to be 44,000 by SDS polyacrylamide gel electrophoresis. The enzyme has a pH optimum for activity at 7.0 and is stable over pH 6.5-9.0. The optimal temperature of the enzyme was 45$\circ$C, and its enzymatic activity was completely inactivated at 55$\circ$C for 30 min. Km value of the enzyme for p-nitrophenyl-$\beta$-D-xylopyranoside was calculated to be 4.6 mM. The effect of various reagents on the $\beta$-xylosidase activity was investigated. The enzyme activity was completely inhibited by Hg$^{2+}$, Cu$^{2+}$ and Zn$^{2+}$. The $\beta$-xylosidase was inactivated by tryptophan-specific reagent, N-bromosuccinimide and tyrosine-specific reagent, iodine. The enzyme could degrade xylo-oligosaccharides to xylose and the enzyme was competitively inhibited by xylose. The $\beta$-xylosidase and endoxylanase from Psedomonas sp. CB-33 hydrolized xylan synergically. The purified enzyme also showed $\alpha$-L-arabinofuranosidase activity.

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Purification and Characterzation of a Restriction Endonuclease from Pseudomonas syringae pv.phaselicola (Pseudomonas syringe pv. phaseolicola로 부터 제한효소의 분리정제 및 특성)

  • Bae, Moo;Lee, Eun-Young
    • Microbiology and Biotechnology Letters
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    • v.22 no.5
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    • pp.485-490
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    • 1994
  • A restriction endonuclease, PsyI, has been isolated from Pseudomonas syringae pv. pha- seolicola, and its catalytic properties have been studied. This enzyme was purified through strepto- mycin sulfate and ammonium sulfate fractionation, phosphocellulose Pll, DEAE-cellulose, hydroxy- apatite and Sephadex G-100 column chromatography. It's molecular weight was about 50,000 dalton as determined by 7.5% polyacrylamide gel electrophoresis containing 0.1% SDS. In catalytic proper- ties, PsyI shows stable at wide ranges of pH between 7.0 and 10.0, of temperature between 30$\circ$C and 37$\circ$C, and its thermal stability is between 25$\circ$C, and 45$\circ$C, at the presence Of 10 mM MgCl$_{2}$-PsyI essentially require Na salt for enzyme reaction, is rather inhibited in the high Na salt concent- ration. The presence of 2-mercaptoethanol is absolutely required for the enzyme activity. This endonuclease, PsyI was determined to be an isoschizomer of SalI from the results of the restriction mapping and DNA sequencing.

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Purification and Characteristics of Amylase from Haloarcular sp. EH-1 (Haloarcular sp. EH-1이 생산하는 Amylase의 정제 및 특성)

  • 정명주;박형숙
    • Microbiology and Biotechnology Letters
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    • v.27 no.2
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    • pp.129-135
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    • 1999
  • EH-1 was highest at 9 days of incubation. This regrowth and enzymatic activity of Haloarcular sp. EH-1 was highest at 9 days of incubation. This amylase was purified by acetone fractionation, DEAG-Cellulose column chromatography, 1st Sephadex G-75 gel filtration, CM-Cellulose column chromatography and 2nd Sephadex G-75 gel filtration. The amylase was purified about 98.64 fold with a yield of 11.75%. The molecular weight of amylase was estimated to be about 43,000and 40,000 by gel filtration and SDS-polyacrylamide gel electrophoresis, respectively, suggesting that the enzyme was a monomer. Amylase had an optimal temperature of 4$0^{\circ}C$, and an optimum pH of 7.0, and the thermal stability was observed the above 50% at 10$0^{\circ}C$ after 1 hour, and the stable range of pH was 6.0 to 8.0. The enzymatic activity was increased in the presence of 10 mM 2-mercaptoethanol, slightly by 10 mM SnCl2.2H2O.FeCl2.4H2O.CuCl2.2H2O.HgCl2.6H2O and SDS. End products from soluble starch were glucose, maltose and maltotriose, and Km value for soluble starch was 2.5mg/ml.

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Experimental studies on the axisymmetric sphere-wall interaction in Newtonian and non-Newtonian fluids

  • Lee, Sang-Wang;Sohn, Sun-Mo;Ryu, Seung-Hee;Kim, Chongyoup;Song, Ki-Won
    • Korea-Australia Rheology Journal
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    • v.13 no.3
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    • pp.141-148
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    • 2001
  • In this research, experimental studies leave been performed on the hydrodynamic interaction between a spherical particle and a plane wall by measuring the force between the particle and wall. To approach the system as a resistance problem, a servo-driving system was set-up by assembling a microstepping motor, a ball screw and a linear motion guide for the particle motion. Glycerin and dilute solution of polyacrylamide in glycerin were used as Newtonian and non-Newtonian fluids, respectively. The polymer solution behaves like a Boger fluid when the concentration is 1,000 ppm or less. The experimental results were compared with the asymptotic solution of Stokes equation. The result shows that fluid inertia plays all important role in the particle-wall interaction in Newtonian fluid. This implies that the motion of two particles in suspension is not reversible even in Newtonian fluid. In non-Newtonian fluid, normal stress difference and viscoelasticity play important roles as expected. In the dilute solution weak shear thinning and the migration of polymer molecules in the inhomogeneous flow field also affect the physic of the problem.

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Differential Induction of Pathogenesis-Related Proteins in the Compatible and Incompatible Interactions of Tomato Leaves with Xanthomonas campestris pv. vesicatoria (Xanthomonas campestris pv. vesicatoria와 토마토잎의 친화적, 불친화적 반응에서 병생성관련 단백질의 유도)

  • 김정동;황병국
    • Korean Journal Plant Pathology
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    • v.11 no.1
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    • pp.53-60
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    • 1995
  • Inoculation with the compatible strain Ds 1 of Xanthomonas campestris pv. vesicatoria caused brownish ad water-soaked lesions, but incompatible strain Bv5-4a produced hypersensitive symptoms with local necrosis on tomato (cv. Kwangyang) leaves. Bacterial populations of the compatible strains Ds 1 propagated more greatly than the incompatible strain Bv5-4a at the frist onset, but no differences were observed 5 days after inoculation. The bacterial infection induced the synthesis and accumulation of soluble proteins in tomato leaves, especially in the incompatible interaction. Native-polyacrylamide gel electrophoresis distinguished the soluble proteins in the tomato leaves infected by the compatible or incompatible strains. A protein of low molecular weight occurred only in the incompatible interaction. Some pathogenesis-related (PR) proteins, especially the 15, 18, 23, 26 and 54 kDa proteins, were detected only in the infected tomato leaves. In the two-dimensional electrophoresis, some proteins with different molecular weights (Mr. 21∼29 kDa) and the pI 8∼9 appeared more distinctly only in the incompatible interaction. These data suggest that the de novo synthesis of some PR proteins in tomato may be significant in defense against X. c. pv. vesicatoria.

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Use of Cationic PAM as a Surface Sizing Additive to Improve Paper Properties

  • Seo, Man-Seok;Lee, Hak-Lae;Youn, Hye-Jung
    • Proceedings of the Korea Technical Association of the Pulp and Paper Industry Conference
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    • 2006.06b
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    • pp.245-250
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    • 2006
  • This study was focused on the use of cationic PAM (Polyacrylamide) as a surface sizing additive to improve the surface sizing properties of paper. Effects of the ionic property, viscosity and charge density of PAM on bending stiffness of surface sized papers were investigated. Use of cationic PAM as a surface sizing additive improved bending stiffness while addition of anionic PAM did not show any effect. Increase of starch holdout with the addition of cationic PAM was attributed as a prime reason of stiffness increase. Viscosity of PAM was one of the most important factors affecting surface sizing due to its influence on the interaction between cationic PAM and oxidized starch solution. Greater improvement of bending stiffness of paper was obtained when high charged PAM was used as an additive. The order of addition was found to have significant influence on the effect of additives since it influences the formation of network structure among starch, cationic PAM, and SA (styrene acrylic acid copolymer). Investigation on the penetration of starch solution was carried out with CLSM (Confocal Laser Scanning Microscopy), and it was shown that the addition of cationic PAM to oxidized starch solution made starch molecules stay on the paper surface rather than penetrating into the paper structure because of the electrostatic interaction between negatively charged fibers and positively charged cationic PAM.

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Haptoglobin as an early indicator of infection in pigs (돼지에서 감염의 조기진단 지표로서의 Haptoglobin 적용에 관한 연구)

  • Moon, Jin-san;Nam, Hyang-mi;Ku, Bok-gyeong;Joo, Yi-seok;Jung, Suk-chan;Kim, Jong-yeom;Park, Yong-ho
    • Korean Journal of Veterinary Research
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    • v.36 no.3
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    • pp.615-619
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    • 1996
  • Sera of pigs with clinically normal and infectious conditions were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) to demonstrate the specific changes in protein profile. In the sera from pigs with infection, haptoglobin with a 40KDa protein was found to be increased as compared to that of sera from normal pigs. As a rapid detection method for monitoring infections at large-scale farms, one of acute phase protein, haptoglobin, was selected to compare the concentrations between infectious and non-infectious conditions. Haptoglobin concentrations were low in pigs with clinically normal conditions but significantly increased in pigs with Aujesky's disease, hog cholera and parvo-virus infection. The studies provide that haptoglobin can be used as an indicator to monitor infections early at farm level.

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