从菠菜叶片中部分纯化赤霉素氧化酶。
Partial purification of gibberellin oxidases from spinach leaves.
机构信息
MSU-DOE Plant Research Laboratory, Michigan State University, East Lansing, Michigan 48824.
出版信息
Plant Physiol. 1987 Sep;85(1):87-90. doi: 10.1104/pp.85.1.87.
Abstract
Four enzyme activities catalyzing the following oxidative steps in the gibberellin (GA) biosynthetic pathway have been extracted from spinach (Spinacia oleracea L.) leaves after exposure to 8 long days: GA(12) --> GA(53) --> GA(44) --> GA(19) --> GA(20). Two of these, GA(53) oxidase and GA(19) oxidase, were separable from the other two, GA(44) oxidase and GA(12) 13-hydroxylase, by anion exchange high performance liquid chromatography (HPLC). Apparent molecular weights of the four enzymes as determined by gel filtration HPLC are: GA(12) 13-hydroxylase, 28,400; GA(53) oxidase, 42,500; GA(44) oxidase, 38,100; GA(19) oxidase, 39,500. GA(44) oxidase was purified approximately 100-fold in 0.3% yield by a combination of ammonium sulfate fractionation, anion exchange HPLC, phenyl-Sepharose chromatography and gel filtration HPLC.
摘要
四种酶活性催化了赤霉素（GA）生物合成途径中的以下氧化步骤，这些酶活性在菠菜（Spinacia oleracea L.）叶片暴露于 8 个长日之后被提取出来：GA(12) --> GA(53) --> GA(44) --> GA(19) --> GA(20)。其中两种，GA(53)氧化酶和 GA(19)氧化酶，可以通过阴离子交换高效液相色谱（HPLC）从另外两种酶，GA(44)氧化酶和 GA(12) 13-羟化酶中分离出来。通过凝胶过滤 HPLC 测定，这四种酶的表观分子量分别为：GA(12) 13-羟化酶，28400；GA(53)氧化酶，42500；GA(44)氧化酶，38100；GA(19)氧化酶，39500。GA(44)氧化酶通过硫酸铵分级沉淀、阴离子交换 HPLC、苯基-Sepharose 色谱和凝胶过滤 HPLC 的组合，以 0.3%的产率被大约纯化了 100 倍。
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