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한문희,성백린,김봉희,민태익,Han, Moon-H.,Seong, Baik-Lin,Kim, Bong-Hee,Mheen, Tae-Ick 생화학분자생물학회 1981 한국생화학회지 Vol.14 No.3
E. coli에서 추출한 penicillin amidase를 황산암모니움에 의한 분별침전 및 DEAE-cellulose, CM-cellulose, Sephadex G-100 컬럼 크로마토그라피에 의해서 3,200배 정제하였고, 이때의 효소의 회수율은 57%이었다. 이것은 기존방법에 비해 비활성은 56배, 그리고 회수율은 1.4배 증가되었다. 이와 같이 얻은 penicillin amidase는polyacrylamide gel 전기영동에 의해 순수함이 증명되었고, SDS-gel 전기영동에 의해 분자량은 55,000임을 알았다. 정제된 효소외 비활성은 기질인 벤질페니실린에 대하여 2.22 ${\mu}$kat/mg protein 이었고 촉매속도항수는 122 $sec^{-1}$이었다. 이 효소의 최적온도 및 pH는 각각 $50^{\circ}C$와 8.5이었으며, $50^{\circ}C$ 이상의 온도와 pH 9.0 이상에서 효소불활성이 빨랐다. $Hg^{++}$를 제외한 대부분의 금속이온은 효소활성에 영향이 없었다. 벤질페니실린에 대한 $K_m$ 상수는 2.9 mM, 기질 저해상수는 35.4 mM, 리고 반응생성물인 6-aminopenicillanic acid (6-APA)와 페닐초산에 의한 저해상수는 각각 10 mM과 4.1 mM이었다. 아실중간체를 형성하는 4단계 반응을 가정하여 steady state 방정식을 유도하고 반응생성물의 효소에 대한 저해양상에 대해서 실험결과 및 보고된 결과들과 비교하였다. Penicillin amidase of Escherichia coli was purified abot 3,200 folds by ammounim sulfate fractionation, DEAE-cellulose, CM-cellulose, and Sephadex G-100 colum chromatography. The overall recovery yield was 57%. As compared with the previous purification method, the purification factor and the recovery yield increased about 56 and 1.4 folds, respectively. The purified enzyme was found to be homogeneous in polyacrplamide gel electrophoresis. The molecular weight determined by SDS-gel electrophoresis was 55,000. The specific activity of penicillin amidase for benzylpenicillin was 2.22 ${\mu}$moles/sec/mg protein and the catalytic rate constant was 122 $sec^{-1}$. The optimum pH and temperature were 8.5 and $50^{\circ}C$, respectively. This enzyme was rapidly deactivated at temperature above $50^{\circ}C$ and pH above 9.0. Almost all of the metal ions did not affect the enzyme activity except for $Hg^{+2}$ ion. The Michaelis constant for benzylpenicillin was 2.9 mM, the substrate inhibition constants for 6-APA and phenylacetic acid were 10 mM and 4.1 mM, respectively A steady state equation based on the four-step ordered reaction with an acyl intermediate was derived. The inhibition modes of both products based on this equation were discussed with respect to the present experimental results.
미생물 페니실린 아미다아제에 관한 연구 ( 3 ) Escherichia coli 가 생산하는 페니실린 아미다아제의 정제 및 동력학적 특성
한문희,성백린,김봉희,민태익 ( Moon H . Han,Baik Lin Seong,Bong Hee Kim,Tae Ick Mheen ) 생화학분자생물학회 1981 BMB Reports Vol.14 No.3
Penicillin amidase of Escherichia coli was purified abot 3,200 folds by ammounim sulfate fractionation, DEAE-cellulose, CM-cellulose, and Sephadex G-100 colum chromatography. The overall recovery yield was 57%. As compared with the previous purification method, the purification factor and the recovery yield increased about 56 and 1.4 folds, respectively. The purified enzyme was found to be homogeneous in polyacrplamide gel electrophoresis. The molecular weight determined by SDS-gel electrophoresis was 55,000. The specific activity of penicillin amidase for benzylpenicillin was 2.22 μmoles/sec/㎎ protein and the catalytic rate constant was 122 sec^(-1). The optimum pH and temperature were 8. 5 and 50℃, respectively, This enzyme was rapidly deactivated at temperature above 50℃ and pH above 9.0. Almost all of the metal ions did not affect the enzyme activity except for Hg^(+2) ion. The Michaelis constant for benzylpenicillin was 2.9 mM, the substrate inhibition constants for 6-APA and phenylacetic acid were 10 mM and 4.1 mM, respectively A steady state equation based on the four-step ordered reaction with an acyl intermediate was derived. The inhibition modes of both products based on this equation were discussed with respect to the present experimental results.
공기청정기와 공기청정장치에 의한 미생물과 바이러스의 제거 효과
김호중(Ho-Jung Kim),이광희(Kwang-Hee Lee),남기석(Ki-Seok Nam),성백린(Baik-Lin Seong),변종길(Jong-Geel Byun),최호선(Ho-Seon Choi) 한국실내환경학회 2004 한국실내환경학회지 Vol.1 No.1
The performance of various air-cleaning devices and an assembled air cleaner has been evaluated for the removal of biological pollutants in indoor air. Bacteria, fungi, and viruses were sprayed in a test chamber, and air samples in the chamber were taken for analysis. Air-cleaning devices - UV lamp, ion generator, an UV LED and plasma electricity dust collector - were tested for their ability in the removal of microorganisms in the air. The UV lamp and the ion generator tested exhbited complete sterilization effect within 4hrs of operation. Other devices were less effective: The extent of removal by the UV LED and the plasma electricity dust collector after 6hrs of operation was about 20% to 82%, depending on the microorganisms tested. The performance of an assembled air cleaner was much superior to individual air-cleaning devices: the extent of removal being 97.6%, 99.1%, 98.7%, and 93.7% for Staphylococcus aureus (Gram(+) bacteria), Escherichia coli (Gram(-) bacteria), Aspergillus niger and Penicillium funiculosum (fungi), respectively, after 3hrs of operation. The removal of influenza virus was even more effective, with 99.9% of removal within 25min of operation. The results show that the air cleaner is effective for the removal of microbial and viral pathogens in the air.