화학공학소재연구정보센터
Protein Expression and Purification, Vol.51, No.1, 126-132, 2007
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 1010bp 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 similar to 37 kDa. Catalytic parameters of the protein were determined using S-D-lactoylglutathione as a thioester substrate. The K-m (61 mu M) 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 OsgyII gene documents its future utility in developing tolerance to various stresses in crop plants. (c) 2006 Elsevier Inc. All rights reserved.