• Title/Summary/Keyword: $E_iC_{50}$

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High-Level Expression of Pseudomonas sp. LBC505 Endoglucanase Gene in Escherichia coli

  • Chun, Sung-Sik;Kim, Yang-Woo;Chung, Young-Chul;Kim, Kyeong-Sook;Sung, Nack-Kie
    • Journal of Microbiology and Biotechnology
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    • v.5 no.1
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    • pp.14-17
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    • 1995
  • Endoglucanase gene of Pseudomonas sp. LBC505 was previously cloned in pUC19 to yield plasmid pLCl. The Pseudomonas sp. LBC505 endoglucanase gene was subcloned in a temperature-regulated Es-cherichia coli expression vector, pAS1, containing the leftward promoter $P_L$ of bacteriophage lambda. The level of gene expression was controlled by the thermal inactivation of the heat-sensitive lambda cI857 repressor. Best yield of endoglucanase was obtained by lowering the incubation temperature to $37^{\circ}C$ after induction at $42^{\circ}C$ for 1h. Under these conditions enzyme production continued for about 5h at a gradually decreasing rate. Ecoli harboring recombinant plasmid pASC10 expressed 4.3 times as much CMCase activity as E.coli containing pLCl. To enhance the expression level of endogl, ucanase gene, we have also changed the presumptive Shine-Dalgamo sequence (AGAGGT) of the gene to consensus sequence (AGGAGGT) by site-directed mutagenesis. The genes mutated were subcloned in pASl resulting in the formation of recombinant plasmid pASS50. E.coli harboring the plasmid pASS50 expressed 6.2-fold higher levels of CMCase activity than that of E.coli harboring pLC1.

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Asymmetric resolution of racemic styrene oxide using recombinant Escherichia coli harboring epoxide hydrolase of Rhodotorula glutinis (Rhodotorula glutinis 유래의 고효율 재조합 Epoxide Hydrolase를 이용한 라세믹 Styrene Oxide의 비대칭 광학분할)

  • Park, Kyu-Deok;Choi, Sung-Hee;Kim, Hee-Sook;Lee, Eun-Yeol
    • KSBB Journal
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    • v.23 no.5
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    • pp.369-374
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    • 2008
  • The effects of reaction temperature and the addition of various detergents on the enantioselective hyrolysis activity of the recombinant Escherichia coli containing the epoxide hydrolase (EH) gene of Rhodotorula glutinis were investigated for the production of enantiopure styrene oxide. The recombinant E. coli harboring the EH gene from R. glutinis exhibited the enantiopreference toward (R)-styrene oxide with the maximum hydrolytic activity of $165.04{\mu}mol/min/mg$ of dry cell weight (dcw). The addition of 0.5% (w/v) Tween 20 at $10^{\circ}C$ increased the initial hydrolysis rate and enantioselectivity by 1.45-fold and 2.0-fold, respectively. Enantiopure (S)-styrene oxide was prepared with 99% ee enantiopurity and 46.0% yield (theoretical yield=50%) from 20 mM racemic styrene oxide.

Studies on the Cellulase produced by Myriococcum of albomyces (Myriococcum albomyces가 생산하는 Cellulase에 관한 연구)

  • Chung, Dong-Hyo
    • Applied Biological Chemistry
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    • v.14 no.1
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    • pp.59-97
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    • 1971
  • As a study on the cellulase of Myriococcum albomyces the culture media for enzyme formation and properties of its crude preparation were investigated and the crude enzyme preparation was further fractionated. The results are summarized as follows: 1. Wheat bran solid culture produced stronger activities of cellulase than rice bran or defatted soy bean meal solid culture. 2. Shaking culture with wheat bran, rice bran or defatted soy bean meal produced higher cellulase activities than solid culture with the corresponding media. 3. The enzyme formation was higher at $45^{\circ}C$ than at $37^{\circ}C$ or $50^{\circ}C$ regardless of the kind of culture medium. 4. The formation of CMCase activity was more promoted by organic nitrogen source than inorganic nitrogen source. 5. The formation of cellulase activities were increased 1.5 to 3.0-fold by adding CMC, Avicel or cellulose powder as an inducer into 5% wheat bran basal medium. 6. Cellulase production using a tank culture procedure with addition of CMC or Avicel as an inducer was the highest at fifth day and thereafter decreased slightly. 7. The crude enzyme preparation showed pH optimum in 4.0 to 4.5, and pH stability in the range of 3.5 to 8.0. Optimum temperature for the activity was $65^{\circ}C$ which was higher than among other cellulases and it was stable at $60^{\circ}C$ for 120 minutes. 8. Dialyzed crude enzyme was activated by $Ca^{++}$ and $Mg^{++}$, but inhibited by $Hg^{++}$, $Cu^{++}$ and $Ag^{+}$. 9. Four different types of cellulase, i. e., fraction I, fraction II-a, fraction II-b, and fraction III were purified from the culture filtrate of Myriococcum albomyces through a sequence of ammonium sulfate fractionation, and elution chromatography on DEAE-Sephadex A-25, Amberlite CG-25 type 2 and hydroxyapatite columns. 10. These four cellulase fractions were showed to be homogenous by electrophoresis and ultracentrifugation and also gave a typical ultraviolet absorption spectrum of protein. 11. Four purified fraction showed different specificity toward substrates, fraction I has a stronger activity toward Avicel, cellulose powder, and gauze than that of other cellulase fractions. Fraction II-a had a powerful activity toward cellobiose but it was almost inactive agaisnt fibrous cellulose contrary to fraction I. On the contrary, the main component fraction II-b had a fairly higher activity on CMC and Avicel. Activity of fraction II-b toward cellobiose was about one-third of that of fraction II-a and activity on Avicel was lower than that of fraction I. Fraction III had a more powerful activity in decreasing viscosity of CMC. 12. Final hydrolysis products of fibrous cellulose by each fraction were cellobiose and glucose. Whereas oligosaccharides were predominant in the early stage of hydrolysis, prolonged reaction produced more glucose than cellobiose. Fraction I and fraction II-a acted synergically on Avicel. 13. Optimum pH for the activities of cellulase fraction I, fraction II-a, fraction II-b and fraction III were found to be 5.5, 5.0, 4.0 and $4.0{\sim}4.5$, respectively. These fractions were found to be stable in the range of pH $3.0{\sim}7.5$. 14. Optimum temperature for the activities of fraction I, fraction II-a, fraction II-b, and fraction III were $50^{\circ}C$, $55^{\circ}C$, $60^{\circ}C$ and $55^{\circ}C$, respectively. No less of activity was found by heating 120 minutes at $55^{\circ}C$ and fraction II-a was more stable than the others at $60^{\circ}C$. 15. Fraction I and fraction II-b were activated by $Ca^{++}$ and $Mg^{++}$ but inhibited by $Hg^{++}$ and $Ag^{+}$.

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Properites of Purified Ascorbate Oxidase in Chlamydomonas reinhardtii (Chlamydomonas reinhardtii 에서 정제한 Ascorbate Oxidase 의 특성)

  • 인용호;이정헌;채영규;최영길;강사욱;정가진;하영칠
    • Korean Journal of Microbiology
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    • v.30 no.3
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    • pp.225-231
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    • 1992
  • l o identil) ;~nclc li~iracterize; In iiscorhate oxiililinp enzyme in ('/rItrn~i~rlon~ir~c~t~itr~~lr.o\ r(1rii. we studicil ;is li)llows. Ascorh;ric oxiiliring cn/;jme activit) f ~ o ~thne crude extract 01' ( ' / ~ l o n ~ ~ . c l o t ~1~~oit~rl~1oin~/.t\ii W;I\ dctccietl by 5pecific active 5ta1ning through nati\e gel cletrophorcsi\ and ~iltra\~iolestp eciroscopy. Ascorb~ttco xidizing c n ~ y m ew i15 partilly 1~1rilieJ by \;~riousp roccclurcs inclucli~lga rnmoniu~ns uIl';~tcp recipit;iion. aJ\orption ~111-om;~togrophy on Iiy~lroxyapaiitca nd Scphailcx <;-I50 gel lillration chrornatogral>liy. Plie ~nolecularw eight 01' the nativc cnrytiic was ahour 88.000 tlalton hy nativc gel elcciroplloresis anci subunit niolecul;ir ~rciglit 55,000 ol' this cnrymc w;~c determined hy SIIS-P.ASI!. The optimum tcmper~tture ii)r the cnrymc nos ahout 5j$^{\circ}$C and pH 4.6 was the optimum. Moreover. ascorhaie oxi~losc in C: reinhardtii was confirmet1 by Ll1e\tcrn blotting technique.

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Purification and Characterization of Xylanase I from Trichoderma koningii ATCC 26113 (Trichoderma koningii ATCC 26113으로부터 Xylanase 1의 순수분리 및 특성)

  • Kim, Hyun-Ju;Kang, Sa-Ouk;Hah, Yung-Chil
    • Korean Journal of Microbiology
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    • v.31 no.1
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    • pp.63-71
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    • 1993
  • A xylanase (xylanase I) was purified 11.9-fold from the culture filtrate of Trichoderma koningii ATCC 26113 by the column chromatography on Sephadex G-75, SP-Sephadex C-50, DEAE-Sephadex A-50 and Sephadex G-50 with an overall yield of 8.2%. The molecular mass determined by gel filtration and sodium dodecyl sulfate polyacrylamide gel electrophoresis was found to be a monomeric polypeptide of ca. 35 kDa. The isoelectric point of the enzyme was estimated to be 9.3. The optimal reaction pH and temperature are 5.8 and 55.deg.C, respectively. The enzyme is stable up to 60.deg.C, while 78% of its activity is lost after the incubation for 10 min at 70.deg.C. The enzyme hydrolyzes sylan with relatively high activity, as well as carboxymethyl cellulose and avicel. The $K_{m}$ values of the enzyme for oat-spelf sylan, larchwood xylan and Avicel were 3.5, 1.6 and 10. 1 mg/ml, respectively. The enzyme hydrolyzed oat-spelt sylan to sylose, sylobiose, sylotriose and arabionoxylobiose, while it degraded larchwood xylan to xylose, xylobiose, xylotriose and arabionoxylobiose, while it degraded larchwood xylan to xylose, xylobiose and xylotriose as the major products. The hydrolysis patterns indicate that xylanase I is endo-enzyme.e.

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Diets with Different Forage/Concentrate Ratios for the Mediterranean Italian Buffalo: In vivo and In vitro Digestibility

  • Fabio, Zicarelli;Calabro, Serena;Piccolo, Vincenzo;D'Urso, Simona;Tudisco, Raffaella;Bovera, Fulvia;Cutrignelli, Monica I.;Infascelli, Federico
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.1
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    • pp.75-82
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    • 2008
  • In vivo and in vitro digestibility of 6 diets with a forage to concentrate ratio (F/C) ranging from 100 to 50:50 (diet 1: all hay, diet 2: 90:10, diet 3: 80:20, diet 4: 70:30, diet 5: 60:40, diet 6: 50:50) were investigated using 6 buffaloes in a $6{\times}6$ Latin square design. For the in vivo trial, the individual faeces of buffaloes were collected 3 times per day for 7 days. Individual pooled faeces and samples of each diet were analysed for chemical composition and insoluble acid ash (AIA) contents in order to estimate the coefficient of apparent digestibility (ADC). On the last day of the in vivo trial a sample of faeces was collected from each animal and used as inoculum for the in vitro test, using the gas production technique (IVGPT). The in vivo organic matter digestibility (ADC) rose as the percentage of concentrate increased up to the 70:30 (F/C) diet (67.01, 73.03, 78.06 and 79.05, respectively for diets 1, 2, 3 and 4); the other two diets (60:40 and 50:50 F/C) unexpectedly did not follow this trend (75.11 and 79.06, respectively for diet 5 and 6). However, these data agree with the results of the in vitro trial. The ADC was positively correlated with the dOM (p<0.001), but not with the gas production at different times; cumulative gas production recorded at the end of incubation (OMCV) showed an irregular trend and was not closely correlated to degraded OM. Estimation of in vivo digestibility from in vitro fermentation data was acceptable, despite leaving room for improvement.

Isolation of Enteropathogenic Escherichia coli, Thermophilic Campylobacter and Salmonellae from Scouring Piglets (설사자돈으로부터 병원성대장균, 캠필로박터속균 및 살모넬라속균의 분리동정)

  • Lee, Ju-hong;Cho, Hee-tack;Kim, Yong-hwan;Kang, Ho-jo;Cha, In-ho
    • Korean Journal of Veterinary Research
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    • v.28 no.1
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    • pp.67-73
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    • 1988
  • This study was conducted to isolate etiological agents from the 103 scouring piglets in Gyeongnam area and also carried out antimicrobial drug susceptibility test and epidemiogical served. The incidence of scouring piglet was most prevalent as 81.6% in the age of 2 to 4 weeks after birth, while the rate was less than 10% in the age of 5 to 6 weeks and under 1 week after birch. When compared the isolation frequency of the each etiological agent, enteropathogenic E. coli was most prevalent as 46.6%, thermophilic Campylobacter 26.2% and Salmonellae was 8.7% in order. In the OK serotyping for 117 isolates of enteropathogenic E. coli, type 0141 : K85 (20.5%), 0157:K88ac(14.5%), 0138:K81 and 0149:K91 (13.3%) were encountered most frequently. In the biotyping for 27 isolates of thermophilic Campylobacter, most strains of C. jejuni were belong to type I (50.0%) and II (25.0%), and most strains of C. coli were belong to biotype I (78.9%). In the serotyping for 9 strains of Salmonellae, 3 strains were grouped as D, 2 strains as C. and each 1 strain was group B and E. The other 2 strains were untypable. The 117 isolates of enteropathogenic E. coli were resistant more than 90% to erythromycin, penicillin, tetracycline and streptomycin, wherease about 90% of the isolates were sensitive to kanamycin and gentamicin. In the case of Salmonellae, all of the isolates were resistant to penicillin, but about 89% of the isolates were sensitive to gentamicin and colistin. All of C. jejuni and C. coli isolates were resistant to cephalothin, but more than 89% of C.jejuni and C. coli were sensitive to kanamycin and gentamicin.

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Particle-in-cell simulation feasibility test for analysis of non-collective Thomson scattering as a diagnostic method in ITER

  • Zamenjani, F. Moradi;Asgarian, M. Ali;Mostajaboddavati, M.;Rasouli, C.
    • Nuclear Engineering and Technology
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    • v.52 no.3
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    • pp.568-574
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    • 2020
  • The feasibility of the particle-in-cell (PIC) method is assessed to simulate the non-collective phenomena like non-collective Thomson scattering (TS). The non-collective TS in the laser-plasma interaction, which is related to the single-particle behavior, is simulated through a 2D relativistic PIC code (XOOPIC). For this simulation, a non-collective TS is emitted from a 50-50 DT plasma with electron density and temperature of ne = 3.00 × 1013 cm-3 and Te = 1000 eV, typical for the edge plasma at ITER measured by ETS system, respectively. The wavelength, intensity, and FWHM of the laser applied in the ETS system are λi,0 = 1.064 × 10-4 cm, Ii = 2.24 × 1017 erg=s·㎠, and 12.00 ns, respectively. The electron density and temperature predicted by the PIC simulation, obtained from the TS scattered wave, are ne,TS = 2.91 × 1013 cm-3 and Te,TS = 1089 eV, respectively, which are in accordance with the input values of the simulated plasma. The obtained results indicate that the ambiguities rising due to the contradiction between the PIC statistical collective mechanism caused by the super-particle concept and the non-collective nature of TS are resolved. The ability and validity to use PIC method to study the non-collective regimes are verified.

A Study on the Effect of Drape by the Construction Method of th eCircular Skirt (Circular Skirt의 구성방법에 따른 Drape효과에 관한 연구)

  • 홍정민
    • Journal of the Korean Society of Costume
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    • v.33
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    • pp.217-228
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    • 1997
  • The purpose of this study was to investigate influence of the construction method of the circular skirt on drapability and shape of static silhouette and length variation. In made 24 types of skirts giving the variations (six kinds of peach skin-like finished fabrics two types of cutting method two types of machine stitch method of hem), The results were as follows: 1. Analysis of drapability In the cutting method thre was similarity between warp direction and true bias direc-tion. In the machine stitch method of hem there was similarity between blind stitch and blind stitch machine. According to the fabrics analysis of drapability was excellent in the order N/P 80/20(fabric 1)>P 100(fabric 5)>P 100(fabric 6)>P 100 (fabric 2)>P 100 (fabric 4)>N/C 50/50(fabric 3). 2. Analysis of the characteristics values of static silhouette shape In the cutting method shape of static sil-houette became wide in the warp direction. In the machine stitch method of hem shape of static silhouette became wide in the blind stitch. According to the fabrics shape of static silhouette became most wide in the N/C 50/50(fabric 3) and shape of static sil-houette become most narrow in the P 100(fabric 2,5) 3. Analysis of the length variation In the cutting method the true bias direc-tion was longer than the warp direction. In the machine stitch method of hem there was simi-machine stitch method of hem there was simi-larity between blind stitch and blind stitch ma-chine. In fabrics the P 100(fabric 6) showed the longest the N/C 50/50(fabric 3) showed most slight. Interaction between the cutting and messure-ment part of skirt lengh. In the warp diretion parts that showed longest length variation were C. G, K, O in the true bias diretion parts that showed most slight length variation were A, E, M, I.

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An Analysis on Thermal Insulation Effect of Farm Structures Coated with Surface Treatment (표면코팅 구조재의 달열효과 분석)

  • Suh, Won-Myung;Yoon, Yong-Cheol
    • Journal of The Korean Society of Agricultural Engineers
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    • v.46 no.4
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    • pp.39-46
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    • 2004
  • This experiment was carried out to study on the effect of surface coating on thermal insulation of farm structures to improve thermal resistance and reflective effect of solar radiation. Nine different types of experimental specimen were compared in the temperature variations of inside and outside; A, B, C, D. E and F types are box container and G, H and I types are drum container. The size of these containers is $1,500{\times}2,000{\times}2,500$ mm and ${\varphi}$ $280{\times}330$ mm, respectively. Specimen of 3-type box(A, B, C) is galvanized steel sheet of thickness 0.45 mm. D, E and F types are sandwich panel of the thickness 50 mm inserted with urethane, glass wool and polystyrene form, respectively. G, H and I types are paint pot using in general. The surface of A. D, E, F and I types didn't any treatment, B, C and G types were treated with thermal insulation coating on the outside surface(B, G) or the inside and outside surface(C). And H type was treated with water paint coating on the only outside surface. In general, the experimental results showed the following tendencies; In case of A, B and C types. it was found that the thermal insulation effect of types coated with thermal insulation coating was improved remarkably than that of no treatment. And the thermal insulation effect between steel sheet and sandwich panel type was nearly similar There was not a significant difference of thermal insulation effect between thermal insulation coating and water paint coating. In time of drum container filled with rough rice, The difference of heat transfer tendency and temperature variation among surface treatments was nearly similar that of box types of galvanized steel sheet. And there was time lag about 6 hours between the temperature of middle part of rice and that of inside or outside surface.