从未发芽大麦中纯化脂氧合酶。特性及产物形成

Purification of a lipoxygenase from ungerminated barley. Characterization and product formation.

作者信息

van Aarle P G, de Barse M M, Veldink G A, Vliegenthart J F

机构信息

Bijvoet Center for Biomolecular Research, Department of Bio-Organic Chemistry, Utrecht University, The Netherlands.

出版信息

FEBS Lett. 1991 Mar 11;280(1):159-62. doi: 10.1016/0014-5793(91)80227-t.

DOI:10.1016/0014-5793(91)80227-t

PMID:1901276

Abstract

Lipoxygenase was purified from ungerminated barley (variety 'Triumph'), yielding an active enzyme with a pI of 5.2 and a molecular mass of approximately 90 kDa. In addition to the 90 kDa band SDS-PAGE showed the presence of two further proteins of 63 kDa. Western blot analysis showed cross-reactivity of each of these proteins with polyclonal antisera against lipoxygenases from pea as well as from soybean, suggesting a close immunological relationship. The 63 kDa proteins appear to be inactive degradation products of the active 90-kDa enzyme. This barley lipoxygenase converts linoleic acid mainly into (9S)-(10E,12Z)-9-hydroperoxy-10,12-octadecadienoic acid, and arachidonic acid into (5S)-(6E,8Z,11Z,14Z)-5-hydroperoxy-6,8,11,14-eic osatetraenoic acid.

摘要

脂氧合酶是从未发芽的大麦（品种“凯旋”）中纯化得到的，得到了一种活性酶，其等电点为5.2，分子量约为90 kDa。除了90 kDa的条带外，SDS-PAGE显示还存在另外两种63 kDa的蛋白质。蛋白质免疫印迹分析表明，这些蛋白质中的每一种都与针对豌豆和大豆脂氧合酶的多克隆抗血清发生交叉反应，这表明它们在免疫学上关系密切。63 kDa的蛋白质似乎是活性90 kDa酶的无活性降解产物。这种大麦脂氧合酶主要将亚油酸转化为(9S)-(10E,12Z)-9-氢过氧-10,12-十八碳二烯酸，将花生四烯酸转化为(5S)-(6E,8Z,11Z,14Z)-5-氢过氧-6,8,11,14-二十碳四烯酸。