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- 저자 : 김승수 ( Hyun Min Choi (검색결과 2 건)
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담배 조직배양에서 6 - Phosphogluconater Dehydrogenase 의 정제와 성질에 관한 연구
최현민,김승수 ( Hyun Min Choi,Soung Soo Kim ) 생화학분자생물학회 1984 BMB Reports Vol.17 No.4
One 6-phosphogluconate dehydrogenase isoenzyme, designated 6PG DH II, was purified from tobacco callus culture line (Nicotiana tobacum L., var. Virginia 115) utilizing ammonium sulfate precipitation, DEAE-cellulose chromatography, Red Sepharose CL-6 B affinity chromatography followed by Sephadex G-100 gel filtration. The purified enzyme was homogeneous according to the criteria of polyacrylamide gel electrophoresis. 6 PG DH II appears to have a dimeric structure with subunit molecular weight of 44,000 and native molecular weight of approximately 83,000. Double reciprocal plot of reaction velocity vs. 6-phosphogluconate concentrations indicated Km values of 17 μM with a marked inhibition of initial velocity at high 6-phosphogluconate concentrations.The NADP^+ double reciprocal plot showed an abrupt increase in the apparent Km and Vmax with increasing NADP^+. The two Km`s for low and high NADP^+ concentrations were 6.5 μM and 70 μM, respectively. The pH optimum of the enzyme was 8.5 in Tris-HCl and 8.0 in phosphate buffer. MgCl₂ was not required for the enzymatic activity. Furthermore, MgCl₂ inhibited the activity at high concentration.
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Choi, Hyun Min,Kim, Soung Soo 연세대학교 자연과학연구소 1984 學術論文集 Vol.13 No.-
담배조직배양의 배지에 indole-3-acetic acid(IAA)와 kinetin을 정상 농도보다 아주 높거나 낮게 첨가한 후 glucose-6-phosphate dehydrogenase 와 6-phosphogluconate dehydrogenase의 활성도와 isoenzyme pattern에 미치는 영향을 조사하였다. 정상 농도보다 훨씬 낮은 IAA농도(10^-8M)와 높은 kinetin 농도(10^-5M)에서는 두 효소의 활성이 대부분의 배양 기간동안 대조군보다 약간 높게 나타났다. 한편 정상 농도보다 아주 높은 IAA 농도(10^-3M)와 낮은 kinetin 농도 (10^-9M)에서는 대부분의 배양 기간동안 대조군보다 낮은 활성을 유지하였다. IAA와 kinetin의 농도 변화에 따른 두 효소의 이러한 활성변화는 전기영동을 통하여 살펴본 두 효소의 isoenzyme pattern 변화에서도 그대로 나타났다. 즉 담배조직배양에서 HMP shunt 조절효소들에 대한 두 hormone의 영향은 서로 상반됨을 알 수 있었다. The effect of hormone stress on the activities of glucose-6-phosphate deydrogenase and 6-phosphogluconate dehydrogenase in culture of Nicotiana tabacum was examined. In the extremely low IAA concentration range (10^-8M) and high kinetin level (10^-5M), the activities of HMP shunt enzymes were enhanced during most culture period as compared to the growth promoting normal IAA and kinetin concentrations. In high toxic IAA concentration range (10^-3M) on the other hand, the level of both enzymes was reduced significantly throughout culture. The slight decrease in enzyme activity also occurred when the tissue was grown on very low kinetin level (10^-9M). The results obtained in the activity profile experiments were also reflected by isoenzyme pattern studies performed at different hormone concentrations.
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