Partial Purification and Characterization of Gibberellin 3β-hydroxylase from Immature Seeds of Phaseolus vulgaris L
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概要
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Gibberellin 3β-hydroxylase,a 2-oxoglutarate-dependent dioxygenase that catalyzes the hy-droxylation of GA_<20> to GA_1, was purified 313-fold from immature seeds of Phaseolus vulgaris L. The mol wt of the enzyme was estimated to be 42,000 by gel filtration HPLC and SDS-poly-acrylamide gel electrophoresis. The enzyme exhibited maximum activity at pH 7.7. The K_m values for [2,3-^3H]GA_<20> and [2,3-^3H]GA_9 were 0.29μM and 0.33 μM, respectively. The enzyme requires 2-oxoglutarate as a cosubstrate; the K_m value for 2-oxoglutarate was 250μM using [^3H]-GA_<20> as a substrate. Fe^<2+> and ascorbate significantly activated the enzyme at all purification steps, while catalase and BSA activated the purified enzyme only. The enzyme was inhibited by divalent cations Mn^<2+>, Co^<2+>, Ni^<2+>, Cu^<2+>, Zn^<2+>, Cd^<2+> and Hg^<2+>. 3β-Hydroxylation of [^3H]-GA_<20> was also inhibited by non-radioactive GA_5, GA_9,GA_<15>, GA_<20> and GA_<44>. The possible site of 3β-hydroxylation in gibberellin biosynthesis is discussed in terms of the substrate specificity of partially purified gibberellin 3β-hydroxylase.
- 日本植物生理学会の論文
著者
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SAKURAI Akira
The Institute of Physical and Chemical Research
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Kamiya Y
Riken Plant Science Center
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Sakurai Akira
The Inst. Of Phys. And Chem. Res.
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Takahashi Nobutaka
The Institute of Physical and Chemical Research
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Kamiya Yuji
The Institute of Physical and Chemical Research
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Kamiya Yuji
Riken Frp.
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Kwak Sang-soo
Department Of Agricultural Chemistry The University Of Tokyo:(present)genetic Engineering Center Kor
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Kamiya Y
Growth Regulation Research Group Riken Plant Science Center
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Kwak S‐s
Korea Res. Inst. Of Bioscience And Biotechnology Taejon Kor
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Kwak Sang-soo
Plant And Animal Cell Technology Research Division Korea Research Institute Of Bioscience
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Sakurai Akira
Fac. Sci. & Eng. Saitama Univ.
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Kamiya Yuji
Frontier Research Program The Institute Of Physical And Chemical Research
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KWAK Sang-Soo
The Institute of Physical and Chemical Research
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Graebe Jan
Pflanzenphysiologisches Institut der Universitat
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Kamiya Yuji
Growth Regulation Research Group Riken Plant Science Center (psc)
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