Efficient purification of cryoprotective dehydrin protein from the radish (Raphanus sativus) taproot

Efficient purification of cryoprotective dehydrin protein from the radish (Raphanus sativus) taproot
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从萝卜 (Raphanus sativus) 主根中高效纯化冷冻保护脱水蛋白

DOI:
10.1007/s00217-014-2228-6
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发表时间:
2014
影响因子:
3.3
通讯作者:
Hermann Wätzig
Hermann Wätzig
中科院分区:
农林科学3区
文献类型:
--
作者:
Masakazu Hara;Saki Uchida;Takae Murata;Hermann Wätzig

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脱水醇是一种与植物耐冷性相关的蛋白质。由于甜菊素显示出有效的冷冻保护活性,因此它具有用于食品储存应用的潜力。本文提出了一种高效纯化萝卜天然脱水素的方法。免疫印迹分析,使用的抗拟南芥KS-型甜菜碱蛋白抗体显示,相关的甜菜碱蛋白积累在萝卜主根。通过两个简单的层析步骤:固定化金属亲和层析和阴离子交换层析,纯化了积累在主根维管组织中的萝卜苷。产率高于先前报道的基于鲜重的产率。结果表明,纯化的白藜芦醇苷对苹果酸脱氢酶的低温保护作用强于小分子冷冻保护剂。这表明,萝卜是一个适当的来源,为生产天然的甜菜碱。
Dehydrin is a protein that is related to cold stress tolerance in plants. Because dehydrin shows potent cryoprotective activity, it has the potential to be used in food storage applications. In this paper, we presented an efficient purification method for native dehydrin from radishes (Raphanus sativus). Immunoblot analysis using an anti-ArabidopsisKS-type dehydrin antibody revealed that the related dehydrin accumulates in the radish taproot. The radish dehydrin that accumulated in the vascular tissues of the taproot was purified through two simple chromatographic steps: immobilized metal affinity chromatography followed by anion exchange chromatography. The yield was higher than yields previously reported on a fresh weight basis. The cryoprotective activity for malate dehydrogenase shown by purified dehydrin was more potent than that shown by small molecule cryoprotectants. This suggests that the radish is an appropriate source for the production of native dehydrin.
DOI: 10.1006/prep.1994.1040
发表时间: 1994-06-01
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