Occurrence of D-serine in rice and characterization of rice serine racemase

被引：25

作者：

Gogami, Yoshitaka ^{[1
]}

Ito, Katsuyoshi ^{[1
]}

Kamitani, Yuji ^{[1
]}

Matsushima, Yuki ^{[1
]}

Oikawa, Tadao ^{[1
]}

机构：

[1] Kansai Univ, Dept Life Sci & Biotechnol, Fac Chem Mat & Bioengn, Osaka 5648680, Japan

来源：

PHYTOCHEMISTRY | 2009年 / 70卷 / 03期

关键词：

Unpolished rice; Oryza sativa L; Gramineae; D-serine; D-amino acid; Serine racemase; Serine dehydratase; Pyridoxal 5 '-phosphate; ALANINE RACEMASE; D-ASPARTATE; BRAIN; ACID; PURIFICATION; BIOSYNTHESIS; ACTIVATION; SEEDLINGS; PROTEIN; CLONING;

D O I：

10.1016/j.phytochem.2009.01.003

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：

Germinated, unpolished rice was found to contain a substantial amount of D-serine, with the ratio of the D-enantiomer to the L-enantiomer being higher for serine than for other amino acids. The relative amount of D-serine (D/(D + L)%) reached approximately 10% six days after germination. A putative serine racemase gene (serr, clone No. 001-110-B03) was found in chromosome 4 of the genomic DNA of Oryza sativa L ssp. Japonica cv. Nipponbare. This was expressed as serr in Escherichia coli and its gene product (SerR) was purified to apparent homogeneity. SerR is a homodimer with a subunit molecular mass of 34.5 kDa, and is highly specific for serine. In addition to a serine racemase reaction, SerR catalyzes D-and L-serine dehydratase reactions, for which the specific activities were determined to be 2.73 and 1.42 nkatal/mg, respectively. The optimum temperature and pH were respectively determined for the racemase reaction (35 degrees C and pH 9.0) and for the dehydratase reaction (35 degrees C and pH 9.5). SerR was inhibited by PLP-enzyme inhibitors. ATP decreased the serine racemase activity of SerR but increased the serine dehydratase activity. Kinetic analysis showed that Mg2+ increases the catalytic efficiency of the serine racemase activity of SerR and decreases that of the serine dehydratase activity. Fluorescence-quenching analysis of the tryptophan residues in SerR indicated that the structure of SerR is distorted by the addition of Mg2+, and this structural change probably regulates the two enzymatic activities. (C) 2009 Elsevier Ltd. All rights reserved.

引用

页码：380 / 387

页数：8

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