title: The multifunctional isopropyl alcohol dehydrogenase of Phytomonas sp. could be the result of a horizontal gene transfer from a bacterium to the trypanosomatid lineage.
abstract: Isopropyl alcohol dehydrogenase (iPDH) is a dimeric mitochondrial alcohol dehydrogenase (ADH), so far detected within the Trypanosomatidae only in the genus Phytomonas. The cloning, sequencing, and heterologous expression of the two gene alleles of the enzyme revealed that it is a zinc-dependent medium-chain ADH. Both polypeptides have 361 amino acids. A mitochondrial targeting sequence was identified. The mature proteins each have 348 amino acids and a calculated molecular mass of 37 kDa. They differ only in one amino acid, which can explain the three isoenzymes and their respective isoelectric points previously found. A phylogenetic analysis locates iPDH within a cluster with fermentative ADHs from bacteria, sharing 74% similarity and 60% identity with Ralstonia eutropha ADH. The characterization of the two bacterially expressed Phytomonas enzymes and the comparison of their kinetic properties with those of the wild-type iPDH and of the R. eutropha ADH strongly support the idea of a horizontal gene transfer event from a bacterium to a trypanosomatid to explain the origin of the iPDH in Phytomonas. Phytomonas iPDH and R. eutropha ADH are able to use a wide range of substrates with similar Km values such as primary and secondary alcohols, diols, and aldehydes, as well as ketones such as acetone, diacetyl, and acetoin. We speculate that, as for R. eutropha ADH, Phytomonas iPDH acts as a safety valve for the release of excess reducing power.
authors.affiliation: Instituto de Biología Molecular y Celular de Rosario-Consejo Nacional de Investigaciones Científicas y Técnicas, Departamento de Microbiología, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Santa Fe, Argentina.
authors.forename: Sara M
authors.initials: SM
authors.lastname: Molinas
authors.forename: Silvia G
authors.initials: SG
authors.lastname: Altabe
authors.forename: Fred R
authors.initials: FR
authors.lastname: Opperdoes
authors.forename: Mark H
authors.initials: MH
authors.lastname: Rider
authors.forename: Paul A M
authors.initials: PA
authors.lastname: Michels
authors.forename: Antonio D
authors.initials: AD
authors.lastname: Uttaro
mesh_terms: D000426:Alcohol Dehydrogenase / Q000737:chemistry* / Q000235:genetics*; D000483:Alleles; D000595:Amino Acid Sequence; D000818:Animals; D015139:Blotting, Southern; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D008401:Gas Chromatography-Mass Spectrometry; D022761:Gene Transfer, Horizontal; D007700:Kinetics; D008969:Molecular Sequence Data; D010802:Phylogeny; D020033:Protein Isoforms; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D014345:Trypanosoma / Q000201:enzymology / Q000235:genetics*; D015032:Zinc / Q000737:chemistry
chemical_list: D020033:Protein Isoforms; D000426:Alcohol Dehydrogenase; D015032:Zinc