title: A functionally critical single nucleotide polymorphism in the gene encoding the membrane-bound alcohol dehydrogenase found in ethanol oxidation-deficient Gluconobacter thailandicus.
abstract: The Gluconobacter thailandicus strains NBRC3254, NBRC3255, NBRC3256, NBRC3257, and NBRC3258 are naturally deficient in the ethanol-oxidizing respiratory chain because they do not produce the cytochrome subunit of the membrane-bound alcohol dehydrogenase (ADH). Draft genomes of G. thailandicus strains NBRC3255 and NBRC3257 indicated that the adhB gene encoding the cytochrome subunit contains four base differences when compared to a closely related gene in the public database One of the nucleotide differences results in an Opal codon at the -19th tryptophan (Trp) in the signal sequence for translocation to the periplasmic space (here, the position of +1st residue is assigned to the N-terminal amino acid residue after signal peptide cleavage), while the other differences result in one missense and two silent amino acid alterations. All five of the G. thailandicus strains were shown to have the Trp(-19)Opal alteration. Ethanol oxidation and ADH activities in NBRC3255 were restored by transformation with a derivative of the endogenous adhB gene, of which the -19th Opal codon was altered to encode Trp. These results indicate that this sequence is a functionally critical single nucleotide polymorphism in the cytochrome subunit. Comparative genomic analyses between the draft genomes of NBRC3255 and NBRC3257 revealed that although the two genomes are closely related, they both have a significant number of unique open reading frames. We suggest that the closely related NBRC3255 and NBRC3257 diverged from a common ancestor having the mutation in the adhB gene, whereas no additional functionally critical mutation occurred in the adhB pseudogene over the course of evolution.
authors.affiliation: Interdisciplinary Graduate Program in Genetic Engineering, The Graduate School, Kasetsart University, Bangkok 10900, Thailand.
authors.forename: Piyanat
authors.initials: P
authors.lastname: Charoenyingcharoen
authors.affiliation: Department of Biological Chemistry, Faculty of Agriculture, Yamaguchi University, Yamaguchi 753-8515, Japan.
authors.forename: Minenosuke
authors.initials: M
authors.lastname: Matsutani
authors.affiliation: Department of Biological Chemistry, Faculty of Agriculture, Yamaguchi University, Yamaguchi 753-8515, Japan; Research Center for Thermotolerant Microbial Resources, Yamaguchi University, Yamaguchi 753-8515, Japan. Electronic address: juji@yamaguchi-u.ac.jp.
authors.forename: Toshiharu
authors.initials: T
authors.lastname: Yakushi
authors.affiliation: Interdisciplinary Graduate Program in Genetic Engineering, The Graduate School, Kasetsart University, Bangkok 10900, Thailand; Department of Microbiology, Faculty of Science, Kasetsart University, Bangkok 10900, Thailand.
authors.forename: Gunjana
authors.initials: G
authors.lastname: Theeragool
authors.affiliation: BIOTEC Culture Collection (BCC), National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Pathumthani 12120, Thailand.
authors.forename: Pattaraporn
authors.initials: P
authors.lastname: Yukphan
authors.affiliation: Department of Biological Chemistry, Faculty of Agriculture, Yamaguchi University, Yamaguchi 753-8515, Japan; Research Center for Thermotolerant Microbial Resources, Yamaguchi University, Yamaguchi 753-8515, Japan.
authors.forename: Kazunobu
authors.initials: K
authors.lastname: Matsushita
mesh_terms: D000426:Alcohol Dehydrogenase / Q000235:genetics*; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D001483:Base Sequence; D020576:Gluconobacter / Q000235:genetics*; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D020641:Polymorphism, Single Nucleotide; D017422:Sequence Analysis, DNA; D013401:Sugar Alcohol Dehydrogenases / Q000235:genetics
chemical_list: D001425:Bacterial Outer Membrane Proteins; D013401:Sugar Alcohol Dehydrogenases; D000426:Alcohol Dehydrogenase; C036131:glycerol oxidase
keywords: Cytochrome c; Gluconobacter; Membrane-bound alcohol dehydrogenase; Pyrroloquinoline quinone; Single nucleotide polymorphism