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Rhizobium japonicum 1-23 Glutamine Synthetase I의 분리 정제 및 성질
이왕식,성하진,방원기,Lee, Wang-Sik,Sung, Ha-Chin,Bang, Won-Gi 생화학분자생물학회 1989 한국생화학회지 Vol.22 No.4
Glycerol를 유일한 탄소원으로 하여, Rhizobium japonicum 1-23으로부터, Glutamine synthetase I (GS I)을 유도하였다. 이 효소를 열처리, 황산암모니움 분별침전, DEAE-Sepharose 음이온 교환, Cibacron Blue F3GA affinity chromatography 및 Ultrogel AcA 22 gel fractionation 과정을 통하여 28.8배 정제하였다. 정제된 GS I의 $\bar{n}$은 4.5이었으며, isoactivity point는 7.53이었다. GS I은 크기가 52,000인 동일한 12개의 subunit으로 구성되어 있는 전체 분자량 약 600,000으로 추정되었다. GS I의 활성은 7.14 mM의 alanine에 의하여 87.2%가 저해되었으며, 5 mM의 inosine에 의하여 98.1%가 저해 되었다. Glutamine에 대한 $K_m$값은 5.4 mM이었으며, $V_{max}$는 $6.90{\mu}mol/min/mg$이었다. Glutamine synthetase I (GS I) was induced in Rhizobium japonicum 1-23 grown on glycerol as a sole source of carbon. The enzyme was purified 28.8 fold by heat-treatment ammonium sulfate fractionation, DEAE-Sepharose ion-exchange chromatography, Cibacron-Blue F3GA affinity chromatography, and Ultrogel AcA 22 gel fractionation. The $\bar{n}$ and the isoactivity point of GS I were 4.5 and 7.53, respectively. Molecular weight of GS I was 600,000 composed of twelve identical 52,000 subunits. The activities of GS I were inhibited by 87.2% (7.14 mM of alanine) and 98.1% (5 mM of inosine). $K_m$ for the substrate, glutamine, was 5.4 mM and $V_{max}$ was $6.90{\mu}mol/min/mg$.
Rhizobium Japonicum 1 - 23 Glutamine Synthetase Ⅰ의 분리 정제 및 성질
이왕식,성하진,방원기 ( Wang Sik Lee,Ha Chin Sung,Won Gi Bang ) 생화학분자생물학회 1989 BMB Reports Vol.22 No.4
Glutamine synthetase I (GS I) was induced in Rhizobium japonicum 1-23 grown on glycerol as a sole source of carbon. The enzyme was purified 28.8 fold by heat-treatment ammonium sulfate fractionation, DEAE-Sepharose ion-exchange chromatography, Cibacron-Blue F3GA affinity chromatography, and Ultrogel AcA 22 gel fractionation. The n and the isoactivity point of GS I were 4.5 and 7.53, respectively. Molecular weight of GS I was 600,000 composed of twelve identical 52,000 subunits. The activities of GS I were inhibited by 87.2% (7.14 mM of alanine) and 98.1% (5 mM of inosine). K_m for the substrate, glutamine, was 5.4 mM and V_(max) was 6.90 μ㏖/min/㎎.
식물성 식품재료로부터 보체계 활성화 다당의 검색 및 그 활성검토
신광순,라경수,성하진,양한철,Shin, Kwang-Soon,Ra, Kyung-Soo,Sung, Ha-Chin,Yang, Han-Chul 한국식품과학회 1993 한국식품과학회지 Vol.25 No.3
식물성 식품재료를 대상으로 보체 용혈분석법$(TCH_{50})$ 을 이용하여 보체 활성화능(항보체 활성)에 대한 검색을 실시하였다. 총 38종의 식용 식물추출물 중 5종의 시료에서 대조구에 비해 60% 이상의 $TCH_{50}$ 감소를 일으키는 비교적 높은 활성을 나타냈으며 그 활성의 순서는 $1000{\mu}g/ml$ 시료 농도에서 생강>토란대>냉이>은행잎>달래이었다. 한편 가장 높은 활성을 보였던 생강에서 조제된 ZR-1의 경우 pronase 소화 후에는 활성의 변화가 없는 반면에 과요오드산 산화에 의해서는 급격한 활성의 감소를 나타냄으로써 ZR-1의 단백질 부위가 아닌 다당 부위가 활성에 기여함을 알 수 있었다. 또한 ZR-1의 항보체 활성은 $Ca^{2+}$ 이온 부재시 부분적인 감소현상을 보였으며, ZR-1을 정상인의 혈청과 반응 후 anti-humanC3를 이용하여 2차원 면역 전기영동을 행한 결과 C3의 분해산물을 관찰할 수 있었다. 또한 이들 획분은 $ACH_{50}$의 저해를 일으켰다. 동 결과로부터 ZR-1의 보체 활성화 양식은 classical pathway 뿐만 아니라 alternative pathway도 경유함을 알 수 있었다. Screenings were performed on edible plants to examine their complement-system activating ability (anti-complementary activity) by hemolytic complement assay $(TCH_{50})$. Among 38 kinds of plant extracts, 5 kinds showed relatively strong anti-complementary activity which decreased $TCH_{50}$ more than 60% comparison with control and the order of activity was Zingiber officinale>Colocasia antiquorum>Capsella bursapastoris>Ginkgo biloba>Alium monanthum in $1000{\mu}g/ml$. The anti-complementary activity of ZR-1 prepared from the root of Zingiber officinale which was showed the most potent activity, did not change by pronase treatment, but decreased greatly by periodate oxidation. These results indicate that not protein moiety but carbohydrate moiety in ZR-1 fraction may also contribute to the anti-complementary activity. Also, the anti-complementary activity of ZR-1 was reduced partially in the absence of the $Ca^{2+}$ ion. When crossed immunoelectrophoresis using anti-human C3 serum was carried out after incubation of normal human serum with the ZR-1 in $Ca^{2+}$ free condition, a cleavage of C3 precipitin line was observed. Furthermore this polysaccharide fraction considerably inhibited $ACH_{50}$. These results also indicate that the mode of complement activation by polysaccharide from Zingiber officinale is via not only the classical pathway but also the alternative pathway.
권미향,임은정,성하진 ( Mee Hyang Kweon,En Jung Lim,Ha Chin Sung ) 한국응용생명화학회 1998 Applied Biological Chemistry (Appl Biol Chem) Vol.41 No.1
During the screening for anti-complementary activity from 10 kinds of edible mushrooms, an alkali extract of Agaricus bisporus showed the highest activity through the complement fixation test. The crude anti-complementary material(AB-O) from Agaricus bisporus was obtained by the alkali extraction using 1 N NaOH containing 5% urea(65℃), followed by methanol reflux, dialysis and lyophilization. The fraction AB-O showed potent anit-complementary and anti-tumor activity against sarcoma-180 injected mice. The fraction AB-O was divided into 5 fractions(AB-20, AB-40, AB-60, AB-80, AB-A) by gradual acetone precipitation. Among them fraction AB-20 having the highest activity and yield was found to contain 39% carbohydrate and 46% protein. The anti-complementary protein-bound polysaccharide AB-20 consisted of glucose, arabinose, xylose, galactose and mannose in a molar ratio of 6.49 : 1.98 : 1.24 : 1.00 : 0.71, respectively and its main component amino acids were alanine(20.59%), isoleucine(16.85%), glutamine+glutamic acid(14.12%) and leucine (13.83%). The anti-complementary activity of AB-20 was decreased greatly by periodate oxidation, but decreased slightly by pronase digestion. This indicates that polysaccharide moiety is corelated with the anti-complementary activity and that protein is also involved in the activity.