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- 저자 : 변시명 ( Kil - Hyun Kim (검색결과 2 건)
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Studies on Soybean Lipoxygenase 2
김길현,변시명,Kim, Kil-Hyun,Byun, Si-Myung 생화학분자생물학회 1981 한국생화학회지 Vol.14 No.2
Lipoxygenase 2 (E.C. 1. 13. 11. 12)를 대두로부터 친화성 크로마토그라피 방법과 DEAE-cellulose chromatography 방법으로 분리, 정제하였다. 친화성 크로마토그라파에서는 흡착제로서 linoleate-aminohexyl Sepharose 4B 를 사용하였으며, pH 6.8 에서 시행한 결과 lipoxygenase 2는 6배 정제되었고 수율은 91%였으나 lipoxygenase 1은 정제도가 오히려 떨어졌으며 수율은 15%이었다. 최종 lipoxygenase 2는 32.4 배 정제되었으며 수율은 28.5%이었다. 여기서 얻은 효소는 전기 영동법에 의해 거의 단일 단백질로 되어있음을 알 수 있었다. 여기서 얻은 lipoxygenase 2를 arachidonic acid와 반응 시킨후 sodium dithionite 로 환원시킨 다음 에테르로 추출한 물질중에 prostaglandin $F_{2{\alpha}}$로 보이는 소량의 물질이 포함되어 있음을 TLC 방법에 의해 확인하였다. Soybean lipoxygenase 2 was purified by the combination of ammonium sulfate fractionation, affinity chromatography on linoleate-aminohexyl Sepharose 4B and DEAE-cellulose chromatography. Purification of lipoxygenase 2 through affinity chromatography resulted in 6-fold purification and the yield of 91%, while lipoxygenase 1 was retrogressed and recovery was only 15%. Overall purification of lipoxygenase 2 was 32.4 folds and the yield was 28.5%, respectively. The purified enzyme preparation was almost homogeneous electrophoretically. When the lipoxygenase 2 preparation was incubated with arachidonic acid, and subsequently reduced with sodium dithionite, a material from ether extract of the reaction mixture seemed to have some of prostaglandin $F_{2{\alpha}}$, which was identified through thin layer chromatography.
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김길현,변시명 ( Kil - Hyun Kim,Si Myung Byun ) 생화학분자생물학회 1981 BMB Reports Vol.14 No.2
Soybean lipoxygenase 2 was purified by the combination of ammonium sulfate fractionation, affinity chromatography on linoleate-aminohexyl Sepharose 4 B and DEAE-cellulose chromatography. Purification of lipoxygenase 2 through affinity chromatography resulted in 6-fold purification and the yield of 91%, while lipoxygenase 1 was retrogressed and recovery was only 15%. Overall purification of lipoxygenase 2 was 32.4 folds and the yield was 28.5%, respectively. The purified enzyme preparation was almost homogeneous electrophoretically. When the lipoxygenase 2 preparation was incubated with arachidonic acid, and subsequently reduced with sodium dithionite, a material from ether extract of the reaction mixture seemed to have some of prostaglandin F_(2α), which was identified through thin layer chromatography.
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