title: Cloning and expression of mitochondrial malate dehydrogenase of Clonorchis sinensis.
abstract: The NAD-dependent mitochondrial malate dehydrogenase (mMDH, EC1.1.1.37) plays pivotal roles in tricarboxylic acid and is crucial for the survival and pathogenecity of parasites. A cDNA, which was identified by high throughput sequencing from the cDNA library constructed from adult Clonorchis sinensis, encoded a putative peptide of 341 amino acids with more than 50% identity with mMDHs from other organisms. The mMDH was expressed in Escherichia coli as the recombinant protein with a GST tag and purified by glutathione-Sepharose 4B column. The recombinant mMDH showed MDH activity of 63.6 U/mg, without lactate dehydrogenase activity and NADPH selectivity. The kinetic constants of recombinant mMDH were determined.
authors.affiliation: Center for Disease Control and Prevention of JiangMen City, Guangdong Province, 529000, People's Republic of China.
authors.forename: Nancai
authors.initials: N
authors.lastname: Zheng
authors.forename: Baoming
authors.initials: B
authors.lastname: Huang
authors.forename: Jin
authors.initials: J
authors.lastname: Xu
authors.forename: Shansheng
authors.initials: S
authors.lastname: Huang
authors.forename: Jinzhong
authors.initials: J
authors.lastname: Chen
authors.forename: Xuchu
authors.initials: X
authors.lastname: Hu
authors.forename: Chaoneng
authors.initials: C
authors.lastname: Ji
authors.forename: Kang
authors.initials: K
authors.lastname: Ying
authors.forename: Xinbing
authors.initials: X
authors.lastname: Yu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003004:Clonorchis sinensis / Q000201:enzymology* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015723:Gene Library; D007700:Kinetics; D008291:Malate Dehydrogenase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D008928:Mitochondria / Q000201:enzymology*; D008969:Molecular Sequence Data; D009243:NAD / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D017422:Sequence Analysis, DNA
chemical_list: D011994:Recombinant Proteins; D009243:NAD; D008291:Malate Dehydrogenase