Characterization and functional validation of glyoxalase II from rice

Glyoxalase II, one of the enzymes of the glyoxalase pathway, cDNA cloned from rice ( OsglyII) consists of 1623 nucleotides with an open reading frame of 1010 bp encoding a polypeptide of 336 amino acids and an estimated isoelectric point of 8.08. The recombinant protein purified from Escherichia col...

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Bibliographic Details
Published in:Protein expression and purification 2007, Vol.51 (1), p.126-132
Main Authors: Yadav, Sudesh Kumar, Singla-Pareek, Sneh L., Kumar, Manoj, Pareek, Ashwani, Saxena, Mukesh, Sarin, Neera B., Sopory, S.K.
Format: Article
Language:English
Subjects:
Abiotic stresses
Abscisic acid
Amino Acid Sequence
Arabidopsis
Characterization
Cloning, Molecular
Enzyme Induction
Escherichia coli
Escherichia coli - enzymology
Functional validation
Glyoxalase II
Molecular Sequence Data
Oryza - enzymology
Oryza sativa
Rice
Sequence Alignment
Thiolester Hydrolases - biosynthesis
Thiolester Hydrolases - chemistry
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Summary:Glyoxalase II, one of the enzymes of the glyoxalase pathway, cDNA cloned from rice ( OsglyII) consists of 1623 nucleotides with an open reading frame of 1010 bp encoding a polypeptide of 336 amino acids and an estimated isoelectric point of 8.08. The recombinant protein purified from Escherichia coli using Ni–NTA affinity chromatography showed molecular mass of ∼37 kDa. Catalytic parameters of the protein were determined using S- d-lactoylglutathione as a thioester substrate. The K m (61 μM) and K cat (301 s −1) values were lower than those reported for Arabidopsis, human and yeast and showed pH optima at 7.2. The E. coli overexpressing OsglyII were able to grow on higher concentration of methylglyoxal. Transcript analysis in rice showed that OsglyII gene expression is stimulated within 15 min in response to various abiotic stresses as well as treatment with abscisic acid or salicylic acid. This multistress response of OsglyII gene documents its future utility in developing tolerance to various stresses in crop plants.
ISSN:1046-5928
1096-0279
DOI:10.1016/j.pep.2006.07.007