title: Identification and characterization of aldehyde dehydrogenase 9 from Lampetra japonica and its protective role against cytotoxicity.
abstract: Aldehyde dehydrogenases (ALDHs), which oxidize aldehyde to corresponding acids, play a major role in the detoxification of various endogenous and exogenous aldehydes. In this study, we cloned and characterized ALDH9 (designated LjALDH9) from Arctic lamprey Lampetra japonica. The open reading frame of LjALDH9 was 1566 bp, encoding 521 amino acids with a predicted molecular mass of 55.68 kDa. LjALDH9 protein had a signal peptide and Aldedh domain with the active site Cys315. In addition, LjALDH9 shares high sequence homology with ALDH9 of jawed vertebrates. Real-time quantitative PCR revealed that LjALDH9 was highly expressed in the buccal gland. A reactive LjALDH9 protein was obtained by prokaryotic expression, two-step-denaturing and refolding and affinity purification. During enzyme activity analysis of recombinant LjALDH9, we found that the most suitable reaction conditions were pH7.0, 16-23 °C and Mn(2+) as the activator. Our study provides theoretical proof that LjALDH9 plays an important role in the parasitic life phase of lamprey.
authors.affiliation: Liaoning Provincial Key Laboratory of Biotechnology and Drug Discovery, Faculty of Life Science, Liaoning Normal University, Dalian 116081, PR China.
authors.forename: Chunhui
authors.initials: C
authors.lastname: Zhao
authors.affiliation: Liaoning Provincial Key Laboratory of Biotechnology and Drug Discovery, Faculty of Life Science, Liaoning Normal University, Dalian 116081, PR China.
authors.forename: Dan
authors.initials: D
authors.lastname: Wang
authors.affiliation: Department of Biotechnology, Dalian Medical University, Dalian 116044, PR China.
authors.forename: Bin
authors.initials: B
authors.lastname: Feng
authors.affiliation: Liaoning Provincial Key Laboratory of Biotechnology and Drug Discovery, Faculty of Life Science, Liaoning Normal University, Dalian 116081, PR China.
authors.forename: Meng
authors.initials: M
authors.lastname: Gou
authors.affiliation: Lamprey Research Center, Liaoning Normal University, Dalian 116029, PR China.
authors.forename: Xin
authors.initials: X
authors.lastname: Liu
authors.affiliation: Lamprey Research Center, Liaoning Normal University, Dalian 116029, PR China. Electronic address: liqw@263.net.
authors.forename: Qingwei
authors.initials: Q
authors.lastname: Li
mesh_terms: D000444:Aldehyde Dehydrogenase / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D000447:Aldehydes / Q000633:toxicity; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003603:Cytotoxins / Q000633:toxicity*; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D007700:Kinetics; D007798:Lampreys / Q000235:genetics*; D008969:Molecular Sequence Data; D009928:Organ Specificity
chemical_list: D000447:Aldehydes; D003603:Cytotoxins; D000444:Aldehyde Dehydrogenase
keywords: ALDH9; Aldehyde dehydrogenase; Enzyme activity analysis; Lampetra japonica