Purification and Properties of NAD-Dependent Malate Dehydrogenase from Mesembryanthemum crystallinum L. Exhibiting Crassulacean Acid Metabolism

Kazuyuki Saitou, Waichi Agata, Takeshi Kawarabata, Yuko Yamamoto, Fumitake Kubota

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

By DEAE-cellulose anion exchange column chromatography, three different forms of NAD-dependent malate dehydrogenase (EC 1.1.1.37) (NAD-MDH) were isolated from leaves of Mesembryanthemum crystallinum L. in the crassulacean acid metabolism mode. NAD-MDH that eluted last from an anion exchange column was purified to a specific activity of 1,096 units (mg protein)-1 using Blue Sepharose CL-6B chromatography. Citrate and adenosine 5’-triphosphate effectively inhibited the activity of NAD-MDH. The inhibition of the enzymatic activity by citrate was reversed by inorganic phosphate and the degree of reversal increased with increasing the concentration of oxaloacetate, the substrate of the reaction. The optimal pH for NAD-MDH activity was around 7.5. Citrate inhibited the enzymatic activity over a wide range of pH and caused a shift in the optimal pH. The enzymatic activity in the presence of citrate was increased by adding inorganic phosphate below pH 7.5. On the basic side, higher than pH 7.5, however, the inhibition by citrate was enhanced by adding inorganic phosphate.

Original languageEnglish
Pages (from-to)760-766
Number of pages7
JournalJapanese Journal of Crop Science
Volume64
Issue number4
DOIs
Publication statusPublished - Jan 1 1995

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Mesembryanthemum
Mesembryanthemum crystallinum
Malate Dehydrogenase
Crassulacean acid metabolism
malate dehydrogenase
Citric Acid
citrates
NAD
Acids
Phosphates
anion exchange
phosphates
Anions
Chromatography
chromatography
Oxaloacetic Acid
DEAE-Cellulose
adenosine triphosphate
agarose
cellulose

All Science Journal Classification (ASJC) codes

  • Food Science
  • Agronomy and Crop Science
  • Genetics

Cite this

Purification and Properties of NAD-Dependent Malate Dehydrogenase from Mesembryanthemum crystallinum L. Exhibiting Crassulacean Acid Metabolism. / Saitou, Kazuyuki; Agata, Waichi; Kawarabata, Takeshi; Yamamoto, Yuko; Kubota, Fumitake.

In: Japanese Journal of Crop Science, Vol. 64, No. 4, 01.01.1995, p. 760-766.

Research output: Contribution to journalArticle

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