High performance liquid chromatography method for the determination of cinnamyl alcohol dehydrogenase activity in soybean roots

被引：26

作者：

dos Santos, W. D. ^{[1
]}

Ferrarese, Maria De Lourdes Lucio ^{[1
]}

Ferrarese-Filho, O. ^{[1
]}

机构：

[1] Univ Maringa, Dept Biochem, Lab Plant Biochem, BR-87020900 Maringa, Parana, Brazil

来源：

PLANT PHYSIOLOGY AND BIOCHEMISTRY | 2006年 / 44卷 / 7-9期

关键词：

cinnamyl alcohol dehydrogenase; coniferaldehyde; high performance liquid chromatography; sinapaldehyde; soybean;

D O I：

10.1016/j.plaphy.2006.08.004

中图分类号：

Q94 [植物学];

学科分类号：

071001 ;

摘要：

This study proposes a simple, quick and reliable method for determining the cinnamyl alcohol dehydrogenase (CAD; EC 1.1.1.195) activity in soybean (Glycine max L. Merr.) roots using reversed-phase high performance liquid chromatography (RP-HPLC). The method includes a single extraction of the tissue and conduction of the enzymatic reaction at 30 degrees C with cinnamaldchydes (coniferyl or sinapyl), substrates of CAD. Disappearance of the substrates in the reaction mixture is monitored at 340 nm (for coniferaldehyde) or 345 nm (for sinapaldehyde) by isocratic elution with methanol/acetic acid through a GLC-ODS (M) column. This HPLC technique furnishes a rapid and reliable measure of cinnamaldehyde substrates, and may be used as an alternative tool to analyze CAD activity in enzyme preparation without previous purification. (c) 2006 Elsevier Masson SAS. All rights reserved.

引用

页码：511 / 515

页数：5

共 12 条

[1] Induced phenylpropanoid metabolism during suberization and lignification: a comparative analysis [J].

Bernards, MA ;

Susag, LM ;

Bedgar, DL ;

Anterola, AM ;

Lewis, NG .

JOURNAL OF PLANT PHYSIOLOGY, 2000, 157 (06) :601-607

[2] Lignin biosynthesis [J].

Boerjan, W ;

Ralph, J ;

Baucher, M .

ANNUAL REVIEW OF PLANT BIOLOGY, 2003, 54 :519-546

[3] Structural and kinetic basis for substrate selectivity in Populus tremuloides sinapyl alcohol dehydrogenase [J].

Bomati, EK ;

Noel, JP .

PLANT CELL, 2005, 17 (05) :1598-1611

[4]

BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3

[5] Metabolite profiling reveals a role for atypical cinnamyl alcohol dehydrogenase CAD1 in the synthesis of coniferyl alcohol in tobacco xylem [J].

Damiani, I ;

Morreel, K ;

Danoun, S ;

Goeminne, G ;

Yahiaoui, N ;

Marque, C ;

Kopka, J ;

Messens, E ;

Goffner, D ;

Boerjan, W ;

Boudet, AM ;

Rochange, S .

PLANT MOLECULAR BIOLOGY, 2005, 59 (05) :753-769

[6] Lignification and related enzymes in Glycine max root growth-inhibition by ferulic acid [J].

dos Santos, WD ;

Ferrarese, MDL ;

Finger, A ;

Teixeira, ACN ;

Ferrarese, O .

JOURNAL OF CHEMICAL ECOLOGY, 2004, 30 (06) :1203-1212

[7] 'Defence lignin' and hydroxycinnamyl alcohol dehydrogenase activities in wounded Eucalyptus gunnii [J].

Hawkins, S ;

Boudet, A .

FOREST PATHOLOGY, 2003, 33 (02) :91-104

[8] The last step of syringyl monolignol biosynthesis in angiosperms is regulated by a novel gene encoding sinapyl alcohol dehydrogenase [J].

Li, LG ;

Cheng, XF ;

Leshkevich, J ;

Umezawa, T ;

Harding, SA ;

Chiang, VL .

PLANT CELL, 2001, 13 (07) :1567-1585

[9] ELICITOR-INDUCED CINNAMYL ALCOHOL-DEHYDROGENASE ACTIVITY IN LIGNIFYING WHEAT (TRITICUM-AESTIVUM L) LEAVES [J].

MITCHELL, HJ ;

HALL, JL ;

BARBER, MS .

PLANT PHYSIOLOGY, 1994, 104 (02) :551-556

[10] Molecular basis for the evolution of xylem lignification [J].

Peter, G ;

Neale, D .

CURRENT OPINION IN PLANT BIOLOGY, 2004, 7 (06) :737-742

← 1 2 →