Purification of active myrosinase from plants by aqueous two-phase counter-current chromatography.
Purification of active myrosinase from plants by aqueous two-phase counter-current chromatography.
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DOI：
10.1002/pca.2535
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发表时间：
2015-01
影响因子：
3.3
通讯作者：
Fahey, Jed W.
中科院分区：
文献类型：
--
作者：
Wade, Kristina L.;Ito, Yoichiro;Ramarathnam, Aarthi;Holtzclaw, W. David;Fahey, Jed W.
黑芥子酶（硫代葡萄糖苷葡萄糖水解酶；E.C. 3.2.1.147）是一种植物酶，引起生物医学界越来越多的兴趣和重要性。黑芥子酶通过水解其丰富的芥子油苷（β-硫代葡萄糖苷 N-羟基硫酸盐）前体，催化异硫氰酸酯的形成，例如萝卜硫素（来自西兰花）和 4-(α-l-吡喃鼠李糖基氧基)苄基异硫氰酸酯（来自辣木），它们是人类细胞保护第 2 相反应的有效诱导剂。开发水性两相逆流色谱 (CCC) 系统，用于快速、三步纯化具有催化活性的黑芥子酶。高浓度磷酸钾和聚乙二醇双相水两相系统 (ATPS) 与新开发的 CCC 配置一起使用，该配置采用螺旋缠绕、平扭管（具有卵形横截面）。直接在 ATPS 中制备初始粗植物提取物并仅注射下层相，可以在螺旋盘 CCC 上进行短暂层析之前对黑芥子酶复合物进行高度选择性的分配。黑芥子酶与其他植物蛋白的最佳相保留和分离提供了 60 倍的纯化。在可预测扩展的 CCC 系统中，从 3 天的西兰花芽、7 天的白萝卜芽、芥菜籽和田间种植的辣木树的叶子中纯化出具有催化活性的黑芥子酶。
Myrosinase (thioglucoside glucohydrolase; E.C. 3.2.1.147), is a plant enzyme of increasing interest and importance to the biomedical community. Myrosinase catalyses the formation of isothiocyanates such as sulforaphane (frombroccoli) and 4-(α-l-rhamnopyranosyloxy)benzyl isothiocyanate (from moringa), which are potent inducers of the cytoprotective phase-2 response in humans, by hydrolysis of their abundant glucosinolate (β-thioglucoside N-hydroxysulphate) precursors. To develop an aqueous two-phase counter-current chromatography (CCC) system for the rapid, three-step purification of catalytically active myrosinase. A high-concentration potassium phosphate and polyethylene glycol biphasic aqueous two-phase system (ATPS) is used with a newly developed CCC configuration that utilises spiral-wound, flat-twisted tubing (with an ovoid cross-section). Making the initial crude plant extract directly in the ATPS and injecting only the lower phase permitted highly selective partitioning of the myrosinase complex before a short chromatography on a spiral disk CCC. Optimum phase retention and separation of myrosinase from other plant proteins afforded a 60-fold purification. Catalytically active myrosinase is purified from 3-day broccoli sprouts, 7-day daikon sprouts, mustard seeds and the leaves of field-grown moringa trees, in a CCC system that is predictably scalable.
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影响因子：
3.6
作者：
Li, Fei;Hullar, Meredith A. J.;Beresford, Shirley A. A.;Lampe, Johanna W.
通讯作者：
Lampe, Johanna W.
DOI：
10.1016/j.jprot.2007.11.006
发表时间：
2008-04-24
影响因子：
--
作者：
Bellostas, Natalia;Petersen, Iben Lykke;Sorensen, Hilmer
通讯作者：
Sorensen, Hilmer
影响因子：
3.1
作者：
Magri, ML;Cabrera, RB;Cascone, O
通讯作者：
Cascone, O
影响因子：
4.1
作者：
Shibusawa, Yoichi;Takeuchi, Naoko;Ito, Yoichiro
通讯作者：
Ito, Yoichiro
影响因子：
5.2
作者：
Li, Yanyan;Zhang, Tao;Li, Xiaoqin;Zou, Peng;Schwartz, Steven J.;Sun, Duxin
通讯作者：
Sun, Duxin