PURIFICATION AND CHARACTERIZATION OF 2 LIPOXYGENASE ISOENZYMES FROM GERMINATING BARLEY

被引：227

作者：

DODERER, A ^{[1
]}

KOKKELINK, I ^{[1
]}

VANDERVEEN, S ^{[1
]}

VALK, BE ^{[1
]}

SCHRAM, AW ^{[1
]}

DOUMA, AC ^{[1
]}

机构：

[1] HEINEKEN TECH BEHEER BV, ZOETERWOUDE, NETHERLANDS

来源：

BIOCHIMICA ET BIOPHYSICA ACTA | 1992年 / 1120卷 / 01期

关键词：

BARLEY; LIPOXYGENASE; PURIFICATION; ISOENZYME; GERMINATION; LINOLEIC ACID HYDROPEROXIDE;

D O I：

10.1016/0167-4838(92)90429-H

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Two lipoxygenase isoenzymes (linoleate:oxygen oxidoreductase, EC 1.13.11.12) present in the embryo of germinating barley seed have been purified to homogeneity and characterized. Both isoenzymes are monomeric proteins with a molecular mass of approx. 90 kDa and crossreact on Western blots with antibodies raised against pea lipoxygenase. They have an apparent K(m) of approx. 16-mu-M for linoleic acid. The isoenzymes differ in the product formed upon incubation with linoleic acid. One of the isoenzymes (lipoxygenase 1) solely forms the 9-HPOD as a product whereas the 13-HPOD is the major product formed by the other isoenzyme (lipoxygenase 2). Lipoxygenase 1 shows a pH-optimum of 6.5, is active in a broad pH range and has an isoelectric point of 5.2-5.3. Lipoxygenase 2 has the same pH optimum, but is active in a narrow pH range and has a significantly higher pI, namely 6.8-6.9. The occurrence of two isoenzymes was confirmed by peptide analysis of the proteins. Amino acid sequence data obtained from proteolytic fragments of lipoxygenase 1 show up to 50% identity with other plant lipoxygenases.

引用

页码：97 / 104

页数：8

共 33 条

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