• 제목/요약/키워드: $E_iC_{50}$

검색결과 564건 처리시간 0.029초

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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    • 제5권1호
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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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Rhodotorula glutinis 유래의 고효율 재조합 Epoxide Hydrolase를 이용한 라세믹 Styrene Oxide의 비대칭 광학분할 (Asymmetric resolution of racemic styrene oxide using recombinant Escherichia coli harboring epoxide hydrolase of Rhodotorula glutinis)

  • 박규덕;최성희;김희숙;이은열
    • KSBB Journal
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    • 제23권5호
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    • pp.369-374
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    • 2008
  • Rhdotorula glutinis epoxide hydrolase 유전자를 pColdI 벡터 와 pET-21b(+) 벡터에 재조합하여 제작한 E. coli를 생촉매로 사용하여 라세믹 styrene oxide에 대하여 회분식 가수분해 반응을 실시하였다. pET-21b(+)/RgEH 재조합 플라스미드 DNA를 가진 E. coli를 $15^{\circ}C$에서 저온 배양할 때 수용성 단백질 형태로 가장 많이 발현되었고, 입체선택적 가수분해 활성과 촉매 안정성이 가장 좋았다. 라세믹 styrene oxide 20 mM에 대하여 반응온도 $30^{\circ}C$에서는 반응시간 20분 동안에 수율 24.0%로 (S)-styrene oxide를 얻은 반면에, 반응온도를 $10^{\circ}C$로 낮추고 0.5% (w/v) Tween 20을 첨가하고 반응시키면 광학순도 99.0% ee 이상의 (S)-styrene oxide을 46.0%의 수율로 얻을 수 있었다. 최적조건에서 E 값은 6.68이었으며, 100 mM의 라세믹 styrene oxide에 대해서는 반응시간 50분에 이론 수율 50% 대비 40%의 높은 수율로 (S)-styrene oxide를 얻을 수 있었다.

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

  • 정동효
    • Applied Biological Chemistry
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    • 제14권1호
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    • pp.59-97
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    • 1971
  • Myriococcum albomycesf가 생산하는 섬유소 분해효소군에 관한 연구로서 호소생산배지 및 조효소의 성질을 규명하고 몇 가지 효소군으로 정제한바 다음과 같은 결과를 얻었다. 1. 밀기울 고채해양의 각 효소 활성은 쌀겨고체배양 및 탈지대두박고체배양의 그것보다 강하였다. 2. 밀기울진탕, 쌀겨진탕배양 및 대두박진탕배양등은 상기의 고체배양보다 각 효소의 활성이 우수하였다. 3. $45^{\circ}C$에서 배양한 것이 배양기의 종류에 관계없이 $37^{\circ}C$$50^{\circ}C$에서 배양한 것보다 각 효소의 활성이 강하였다. 4. CMCase는 무기 질소원보다 유기 질소원을 첨가하므로서 더욱 생성이 촉진되었다. 5. 기본밀기울진탕배양기에 CMC, Avicel, 여지분말 등의 indncer를 첨가 하므로서 각 효소활성은 $1.5{\sim}3$배나 증가되었다. 6. CMC와 Avicel을 inducer로 하여 jar formentor에서 배양할 때 작 효소의 활성은 대개 5일째에 최고에 달하였다. 7. Cellulae 조효소의 최적 pH는 $4.0{\sim}4.5$, pH안정성은 $3.5{\sim}8.0$이였다. 그리고 최적온도는 $65^{\circ}C$ 부근으로 다른 사상균의 cellulase에 비하여 높으며 온도안정성도 $60^{\circ}C$에서 120분으로 거의 실활되지 않았다. 8. 조효소의 활성은 $Ca^{++}$, $Mg^{++}$으로 부활되며, $Hg^{++}$, $Cu^{++}$, $Ag^{+}$는 강한 저해체였다. 그리고 투석으로 약간 그 활성이 저하되었다. 9. 배양액을 여과하고 황산암모니움 분획, DEA-E-sephadex A-25, Amberlite CG-50 및 hydroxy-apatite column chromatography로 Avicel, CMC, 여지 분말에 대하여 활성이 다른 4개의 fraction을 분리 하고 이를 cellulase fraction I, fraction II-a, fraction II-b 및 fraction III라고 명명하였다. 10. 이들 4개의 fraction은 전기 영동, 초원심상 및 자외선흡수 등으로 보아서 단일의 단백질로 생각되었다. 11. Fraction I은 Avicelase활성이 강하고, fraction II-a는 cellobiase 활성이 강하였다. 그리고 fraction II는 CMCase 활성이 강하였으며, fraction III는 CMC 점도감소 활성이 강하였다. 12. 섬유소질을 각 fraction으로 가수분해한 최종산물은 cellobiose 및 glucose였다. 그리고 fraction I과 fraction II-a는 Avicel을 협동적으로 분해하였다. 13. Fraction I의 최적 pH 5.5, fraction II-a는 pH 5.0, fraction II-b는 pH 4.0, fractionIII는 pH $4.0{\sim}4.5$이며, 각 fraction의 pH 안정성은 pH $3.0{\sim}7.0$이였다. 14. Fraction I의 최적온도는 $50^{\circ}C$, fractionII-a는 $55{\sim}60^{\circ}C$, fraction II-b 는 $60^{\circ}C$, fraction III는 $55^{\circ}C$이며 각 fraction의 열안정성은 $55^{\circ}C$ 부근에서 120분으로 거의 실활되지 않고 fraction II-a는 $60^{\circ}C$에서도 특별히 안정하였다. 15. Fraction I과 fraction II-b 활성은 $Ag^{++}$, $Hg^{++}$에 의하여 저해되며 $Ca^{++}$, $Mg^{++}$으로는 부활되었다.

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

  • 인용호;이정헌;채영규;최영길;강사욱;정가진;하영칠
    • 미생물학회지
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    • 제30권3호
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    • pp.225-231
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    • 1992
  • 생물체내의 ascorbic acid :dehydroascorbic acid system 을 조절하는데 관여하는 것으로 알려진 ascorbate 산화효소의 활성을 단세포 녹조류인 Chlamydomonas reinhardtii 에서 확인함으로써 이 효소의 존재와 특성을 고찰하였다. 조효소는 native gel 전기 영동에 의한 활성 염색과 분광광도계에 의한 ascorbic acid .의 흡광도 감소 측정에서 뚜렷한 활성을 나타내었다. 황산암모늄 침전, hydroxyapatite 흡착 크로마토그래피, 그리고 Sephadex G-150 gel 여과 크로마토그래피 분리한 결과 26.1 units/mg /의 specific activity 를 나타내었다. 분리된 ascorbate 산화효소는 $55^{\circ}C$, pH 4.6 에서 가장 활성도가 높았다. Native gel 전기 영동에서 88,000 dalton 갸량의 분자량을 가지고 있었고 소단위체의 분자량을 55,000 이었다. 또한 Western bloting technique 을 이용하여 C. reinhardtii 의 ascorbate oxidase를 확인하였다.

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

  • Kim, Hyun-Ju;Kang, Sa-Ouk;Hah, Yung-Chil
    • 미생물학회지
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    • 제31권1호
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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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    • 제21권1호
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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.

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

  • 이주홍;조희택;김용환;강호조;차인호
    • 대한수의학회지
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    • 제28권1호
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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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    • 제52권3호
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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.

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

  • 홍정민
    • 복식
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    • 제33권
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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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표면코팅 구조재의 달열효과 분석 (An Analysis on Thermal Insulation Effect of Farm Structures Coated with Surface Treatment)

  • 서원명;윤용철
    • 한국농공학회논문집
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    • 제46권4호
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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.