Molecular and biochemical characterization of a serine racemase from Arabidopsis thaliana

被引:48
作者
Fujitani, Y
Nakajima, N
Ishihara, K
Oikawa, T
Ito, K
Sugimoto, M
机构
[1] Okayama Univ, Bioresources Res Inst, Kurashiki, Okayama 7100046, Japan
[2] Okayama Prefectural Univ, Collaborat Res Ctr, Okayama 7191197, Japan
[3] Okayama Univ Sci, Dept Life Sci, Okayama 7000005, Japan
[4] Kansai Univ, Fac Engn, Dept Biotechnol, Suita, Osaka 5648680, Japan
[5] Sapporo Breweries Ltd, Plant Bioengn Res Labs, Ohta, Gunma 3700393, Japan
关键词
Arabidopsis thaliana; serine racemase; D-amino acid; pyridoxal 5 '-phosphate;
D O I
10.1016/j.phytochem.2006.01.003
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
A cDNA encoding a homolog of mammalian serine racemase, a unique enzyme in eukaryotes, was isolated from Arabidopsis thaliana and expressed in Escherichia coli cells. The gene product, of which the amino acid residues for binding pyridoxal 5'-phosphate (PLP) are conserved in this as well as mammalian serine racemases, catalyzes not only serine racemization but also dehydration of serine to pyruvate. The enzyme is a homodimer and requires PLP and divalent cations, Ca2+, Mg2+, Mn2+, Fe2+, or Ni2+, at alkaline pH for both activities. The racemization process is highly specific toward L-serine, whereas L-alanine, L-arginine, and L-glutamine were poor substrates. The V-max/K-m values for racemase activity of L- and D-serine are 2.0 and 1.4 nmol/mg/min/mM, respectively, and those values for L- and D-serine on dehydratase activity are 13 and 5.3 nmol/mg/min/mM, i.e. consistent with the theory of racemization reaction and the specificity of dehydration toward L-serine. Hybridization analysis showed that the serine racemase gene was expressed in various organs of A. thaliana. (c) 2006 Elsevier Ltd. All rights reserved.
引用
收藏
页码:668 / 674
页数:7
相关论文
共 32 条
[1]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[2]   A note on the kinetics of enzyme action. [J].
Briggs, GE ;
Haldane, JBS .
BIOCHEMICAL JOURNAL, 1925, 19 (02) :338-339
[3]   Chromatographic determination of L- and D-amino acids in plants [J].
Brückner, H ;
Westhauser, T .
AMINO ACIDS, 2003, 24 (1-2) :43-55
[4]   Direct calcium binding results in activation of brain serine racemase [J].
Cook, SP ;
Galve-Roperh, I ;
del Pozo, AM ;
Rodríguez-Crespo, I .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2002, 277 (31) :27782-27792
[5]   Human serine racemase: moleular cloning, genomic organization and functional analysis [J].
De Miranda, J ;
Santoro, A ;
Engelender, S ;
Wolosker, H .
GENE, 2000, 256 (1-2) :183-188
[6]   Cofactors of serine racemase that physiologically stimulate the synthesis of the N-methyl-D-aspartate (NMDA) receptor coagonist D-serine [J].
de Miranda, J ;
Panizzutti, R ;
Foltyn, VN ;
Wolosker, H .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2002, 99 (22) :14542-14547
[7]   THE PRESENCE OF FREE D-ASPARTIC ACID IN RODENTS AND MAN [J].
DUNLOP, DS ;
NEIDLE, A ;
MCHALE, D ;
DUNLOP, DM ;
LAJTHA, A .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1986, 141 (01) :27-32
[8]   Chromatographic determination of amino acid enantiomers in beers and raw materials used for their manufacture [J].
Erbe, T ;
Brückner, H .
JOURNAL OF CHROMATOGRAPHY A, 2000, 881 (1-2) :81-91
[9]   BIOSYNTHETIC ALANINE RACEMASE OF THE SALMONELLA-TYPHIMURIUM - PURIFICATION AND CHARACTERIZATION OF THE ENZYME ENCODED BY THE AIR GENE [J].
ESAKI, N ;
WALSH, CT .
BIOCHEMISTRY, 1986, 25 (11) :3261-3267
[10]  
ESAKI N, 2002, ENZYME CATALYSIS ORG, V3, P1281