Characterization of a Functionally Active Recombinant1-Deoxy-D-Xylulose-5-Phosphate Synthase from Babesiabovis

[摘要] References(33)The1-deoxy-D-xylulose-5-phosphate synthase (DXS) enzyme has been characterized in otherspecies, but not in the genus Babesia, which causes major losses in thelivestock industries worldwide. Therefore, we isolated, cloned and expressed the wild-typeB. bovis dxs cDNA in Escherichia coli and evaluatedits enzymatic activity in vitro. DNA sequence analysis revealed an openreading frame of 2061 bp capable of encoding a polypeptide of 686 amino acid residues witha calculated isoelectric point of pH 6.93 and a molecular mass of 75 kDa. The expressedsoluble recombinant fusion DXS protein was approximately 78 kDa, which is similar to thenative enzyme identified from the parasite merozoite using anti-rDXS serum. Therecombinant fusion DXS enzyme exhibited Km values of 380 ± 46µM and 790 ± 52 µM for D,L-glyceraldehyde 3-phosphateand pyruvate, respectively. In this work, we present the first cloning, expression andcharacterization of DXS enzyme from B. bovis.

[学科分类] 兽医学

[关键词] Babesia bovis;characterize;DXS;isoprenoid biosynthesis