Yuzo Yamada

Genera and species in acetic acid bacteria

Taxonomic studies of acetic acid bacteria were historically surveyed. The genus Acetobacter was first introduced in 1898 with a single species, Acetobacter aceti. The genus Gluconobacter was proposed in 1935 for strains with intense oxidation of glucose to gluconic acid rather than oxidation of ethanol to acetic acid and no oxidation of acetate. The genus Acetomonas was described in 1954 for strains with polar flagellation and no oxidation of acetate. The proposals of the two generic names were due to confusion, and Acetomonas was a junior subjective synonym of Gluconobacter. The genus Acetobacter was in 1984 divided into two subgenera, Acetobacter and Gluconoacetobacter. The latter was elevated to the genus Gluconacetobacter in 1998. In the acetic acid bacteria, ten genera are presently recognized and accommodated to the family Acetobacteraceae, the Alphaproteobacteria: Acetobacteer, Gluconobacter, Acidomonas, Gluconacetobacter, Asaia, Kozakia, Swaminathania, Saccharibacter, Neoasaia and Granulibacter. In contrast, the genus Frateuria, strains of which were once named pseudacetic acid bacteria, was classified into the Gammaproteobacteria. The genus Gluconacetobacter was phylogenetically divided into two groups: the Gluconacetobacter liquefaciens group and the Gluconacetobacter xylinus group. The two groups were discussed taxonomically.

Gluconobacter japonicus sp. nov., an acetic acid bacterium in the Alphaproteobacteria.

Five strains, NBRC 3271(T), NBRC 3272, NBRC 3263, NBRC 3260 and NBRC 3269 were examined genetically, phylogenetically, phenotypically and chemotaxonomically. The DNA G+C contents of the five strains were 55.1-56.4 mol%. The five strains had low levels of DNA-DNA hybridization of 13-51 % to the type strains of Gluconobacter frateurii, Gluconobacter thailandicus, Gluconobacter oxydans, Gluconobacter cerinus, Gluconobacter albidus and Gluconobacter kondonii and formed a cluster that was separate from the type strains of the six Gluconobacter species given above in phylogenetic trees based on 16S rRNA gene and 16S-23S rRNA gene internal transcribed spacer sequences. The five strains weakly produced dihydroxyacetone from glycerol, but not 2,5-diketo-d-gluconate or a water-soluble brown pigment from d-glucose and contained ubiquinone-10. The five strains were assigned as representing a novel species of the genus Gluconobacter, for which the name Gluconobacter japonicus sp. nov. is proposed. The type strain is NBRC 3271(T) (=BCC 14458(T)=strain 7(T), K. Kondo). Cells of the type strain are motile by means of polar flagella and the DNA G+C content is 56.4 mol%.

Tanticharoenia sakaeratensis gen. nov., sp. nov., a New Osmotolerant Acetic Acid Bacterium in the α-Proteobacteria

Tanticharoenia sakaeratensis gen. nov., sp. nov. is proposed for three strains isolated from soil collected in Thailand. The three strains, AC37T, AC38, and AC39, were included within a lineage comprising the genera Asaia, Kozakia, Swaminathania, Neoasaia, Acetobacter, Gluconobacter, and Saccharibacter in a phylogenetic tree based on 16S rRNA gene sequences, but formed a quite different, independent cluster. Pair-wise sequence similarities of strain AC37T were 96.5–92.1% to the type strains of Acetobacter aceti, Gluconobacter oxydans, Acidomonas methanolica, Gluconacetobacter liquefaciens, Asaia bogorensis, Kozakia baliensis, Swaminathania salitolerans, Saccharibacter floricola, Neoasaia chiangmaiensis, and Granulibacter bethesdensis. The three strains had DNA base compositions comprising respectively 65.6, 64.5, and 65.6 mol % G+C with a range of 1.1 mol %, and formed a single species. Phenotypically, the three strains did not oxidize acetate or lactate, but grew on 30% D-glucose (w/v). Chemotaxonomically, they had Q-10. The type strain is AC37T (= BCC 15772T = NBRC 103193T).

Asaia lannaensis sp. nov., a New Acetic Acid Bacterium in the Alphaproteobacteria

Asaia lannaensis sp. nov. was described for two strains isolated from flowers of the spider lily collected in Chiang Mai, Thailand. The isolates produced acetic acid from ethanol on ethanol/calcium carbonate agar, differing from the type strains of Asaia bogorensis, Asaia siamensis, and Asaia krungthepensis, but did not grow in the presence of 0.35% acetic acid (v/v). The new species is the fourth of the genus Asaia, the family Acetobacteraceae.

Neokomagataea gen. nov., with Descriptions of Neokomagataea thailandica sp. nov. and Neokomagataea tanensis sp. nov., Osmotolerant Acetic Acid Bacteria of the α-Proteobacteria

Isolates AH11T and AH13T were isolated from flowers of lantana and candle bush respectively collected in Thailand. In phylogenetic trees based on 16S rRNA gene sequences, the two isolates formed an independent cluster, which was then connected to the type strain of Saccharibacter floricola. The calculated pair-wise 16S rRNA gene sequence similarities of isolate AH11T were 95.7–92.3% to the type strains of the type species of the 12 genera of acetic acid bacteria. The DNA base composition was from 51.2 to 56.8 mol % G+C, with a range of 5.6 mol %. When isolate AH11T was labeled, DNA-DNA similarities were 100, 12, 4, 5, and 4% respectively to isolates AH11T and AH13T and the type strains of Saccharibacter floricola, Gluconobacter oxydans, and Acetobacter aceti. The two isolates were non-motile and did not oxidize either acetate or lactate. No growth was found in the presence of 0.35% acetic acid w/v. The two isolates were not osmophilic but osmotolerant, produced 2,5-diketo-D-gluconate from D-glucose, and did not oxidize lactate, thus differing from strains of Saccharibacter floricola, which showed weak lactate oxidation. The two isolates contained unsaturated C18:1ω7c fatty acid as the major fatty acid, and were unique in the presence of a considerable amount of straight-chain C18:12OH fatty acid. Q-10 was present as the major isoprenoid quinone. Neokomagataea gen. nov. was proposed with the two species, Neokomagataea thailandica sp. nov. for isolate AH11T (=BCC 25710 T =NBRC 106555T), which has 56.8 mol % G+C, and Neokomagataea tanensis sp. nov. for isolate AH13T (=BCC 25711T=NBRC 106556T), which has 51.2 mol % G+C.

Ameyamaea chiangmaiensis gen. nov., sp. nov., an Acetic Acid Bacterium in the α-Proteobacteria

Two isolates, AC04T and AC05, were isolated from the flowers of red ginger collected in Chiang Mai, Thailand. In phylogenetic trees based on 16S rRNA gene sequences, the two isolates were included within a lineage comprised of the genera Acidomonas, Gluconacetobacter, Asaia, Kozakia, Swaminathania, Neoasaia, Granulibacter, and Tanticharoenia, and they formed an independent cluster along with the type strain of Tanticharoenia sakaeratensis. The calculated pair-wise sequence similarities of isolate AC04T were 97.8–92.5% to the type strains of the type species of the 11 genera of acetic acid bacteria. The DNA base composition was 66.0–66.1 mol % G+C with a range of 0.1 mol %. A single-stranded, labeled DNA from isolate AC04T presented levels of DNA-DNA hybridization of 100, 85, 4, and 3% respectively to DNAs from isolates AC04T and AC05 and the type strains of Tanticharoenia sakaeratensis and Gluconacetobacter liquefaciens. The two isolates were unique morphologically in polar flagellation and physiologically in intense acetate oxidation to carbon dioxide and water and weak lactate oxidation. The intensity in acetate oxidation almost equaled that of the type strain of Acetobacter aceti. The two isolates had Q-10. Isolate AC04T was discriminated from the type strains of the type species of the 11 genera by 16S rRNA gene restriction analysis using restriction endonucleases TaqI and Hin6I. The unique phylogenetic, genetic, morphological, physiological, and biochemical characteristics obtained indicate that the two isolates can be classified into a separate genus, and Ameyamaea chiangmaiensis gen. nov., sp. nov. is proposed. The type strain is isolate AC04T (=BCC 15744T, =NBRC 103196T), which has a DNA G+C content of 66.0 mol %.