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 coli using Ni-NTA affinity chromatography showed molecular mass of approximately 37 kDa. Catalytic parameters of the protein were determined using S-D-lactoylglutathione as a thioester substrate. The K(m) (61 microM) 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.
Yadav
Sudesh Kumar
SK
Plant Molecular Biology Group, International Centre for Genetic Engineering and Biotechnology, New Delhi 110 067, India.
Singla-Pareek
Sneh L
SL
Kumar
Manoj
M
Pareek
Ashwani
A
Saxena
Mukesh
M
Sarin
Neera B
NB
Sopory
S K
SK
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D004790:Enzyme Induction; D004926:Escherichia coli / Q000201:enzymology; D008969:Molecular Sequence Data; D012275:Oryza / Q000201:enzymology*; D016415:Sequence Alignment; D013869:Thiolester Hydrolases / Q000096:biosynthesis / Q000737:chemistry*; D000099301:Hydroxyacylglutathione Hydrolase
D013869:Thiolester Hydrolases; D000099301:Hydroxyacylglutathione Hydrolase