title: Characterization of a functionally active recombinant 1-deoxy-D-xylulose-5-phosphate synthase from Babesia bovis.
abstract: The 1-deoxy-D-xylulose-5-phosphate synthase (DXS) enzyme has been characterized in other species, but not in the genus Babesia, which causes major losses in the livestock industries worldwide. Therefore, we isolated, cloned and expressed the wild-type B. bovis dxs cDNA in Escherichia coli and evaluated its enzymatic activity in vitro. DNA sequence analysis revealed an open reading frame of 2061 bp capable of encoding a polypeptide of 686 amino acid residues with a calculated isoelectric point of pH 6.93 and a molecular mass of 75 kDa. The expressed soluble recombinant fusion DXS protein was approximately 78 kDa, which is similar to the native enzyme identified from the parasite merozoite using anti-rDXS serum. The recombinant fusion DXS enzyme exhibited Km values of 380 ± 46 µM and 790 ± 52 µM for D,L-glyceraldehyde 3-phosphate and pyruvate, respectively. In this work, we present the first cloning, expression and characterization of DXS enzyme from B. bovis.
authors.affiliation: Lanzhou Institute of Husbandry and Veterinary Pharmaceutical Science, Chinese Academy of Agricultural Sciences, Key Laboratory for Veterinary Drug Innovation, Ministry of Agriculture, Lanzhou 730050, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Wang
authors.forename: You-Ming
authors.initials: YM
authors.lastname: Shen
authors.forename: Bing
authors.initials: B
authors.lastname: Li
authors.forename: Xu-zheng
authors.initials: XZ
authors.lastname: Zhou
authors.forename: Cui-cui
authors.initials: CC
authors.lastname: Liu
authors.forename: Ji-yu
authors.initials: JY
authors.lastname: Zhang
mesh_terms: D000595:Amino Acid Sequence; D016793:Babesia bovis / Q000201:enzymology*; D003001:Cloning, Molecular; D019295:Computational Biology; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D008969:Molecular Sequence Data; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D014166:Transferases / Q000235:genetics / Q000378:metabolism*
chemical_list: D015800:Protozoan Proteins; D014166:Transferases; C109461:deoxyxylulose-5-phosphate synthase
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