Asaia krungthepensis sp. nov., an acetic acid bacterium in the α-Proteobacteria

Abstract

Three bacterial strains were isolated from flowers collected in Bangkok, Thailand, by an enrichment-culture approach for acetic acid bacteria. Phylogenetic analysis based on 16S rRNA gene sequences showed that the isolates were located in the lineage of the genus Asaia but constituted a cluster separate from the type strains of Asaia bogorensis and Asaia siamensis. The DNA base composition of the isolates was 60?2-60?5 mol% G+C, with a range of 0?3 mol%. The isolates constituted a taxon separate from Asaia bogorensis and Asaia siamensis on the basis of DNA-DNA relatedness. The isolates had morphological, physiological, biochemical and chemotaxonomic characteristics similar to those of the type strains of Asaia bogorensis and Asaia siamensis, but the isolates grew on maltose. The major ubiquinone was Q 10 . On the basis of the results obtained, the name Asaia krungthepensis sp. nov. is proposed for the isolates. The type strain is isolate AA08 T (=BCC 12978 T =TISTR 1524 T =NBRC 100057 T =NRIC 0535 T ), which had a DNA G+C content of 60?3 mol% and was isolated from a heliconia flower ('paksaasawan' in Thai; Heliconia sp.) collected in Bangkok, Thailand.The genus Asaia was introduced with a single species, Asaia bogorensis, as the fifth genus of the family Acetobacteraceae (Yamada et al., 2000). Asaia siamensis, the second species, was described by Katsura et al. (2001). Strains assigned to the genus Asaia are characterized by poor or non-existent production of acetic acid from ethanol and by the absence of growth in the presence of 0?35 % acetic acid (v/v). On the other hand, strains of the genera Acetobacter, Gluconobacter, Acidomonas, Gluconacetobacter and Kozakia produce large amounts of acetic acid from ethanol and grow in the presence of 0?35 % acetic acid. According to Yamada et al. (2000) and Katsura et al. (2001), strains of the genus Asaia have been isolated mostly from tropical flowers. During the course of our studies on acetic acid bacteria isolated from flowers collected in Thailand, we found that three isolates constitute a novel species; this paper describes Asaia krungthepensis sp. nov.Three bacterial strains were isolated from heliconia flowers ('paksaasawan' in Thai; Heliconia sp.) collected in Bangkok, Thailand, by an enrichment-culture approach using a sucrose medium, which was composed of 2?0 % sucrose, 0?3 % peptone, 0?3 % yeast extract and 0?3 % acetic acid (v/v) and adjusted to pH 3?5. When bacterial growth was found in the sucrose medium, micro-organisms were streaked on agar plates containing 2?0 % D-glucose, 0?5 % ethanol (v/v), 0?3 % peptone, 0?3 % yeast extract, 0?7 % CaCO 3 and 1?2 % agar (w/v). These acetic acid bacteria were selected as acid-producing bacterial strains that formed clear zones around colonies on agar plates. The three isolates, AA08 T (=BCC 12978 T ), AA09 (=BCC 12979) and AA06 (=BCC 12977), were cultured at 30 uC on agar slants comprising glucose/ethanol/CaCO 3 /agar medium (GECA medium) that contained 2?5 % D-glucose, 0?5 % ethanol, 3Present address: 2-3-21 Seinan...

Neoasaia chiangmaiensis gen. nov., sp. nov., a novel osmotolerant acetic acid bacterium in the .ALPHA.-Proteobacteria

“…An acetic acid bacterium, designated as isolate AC28 T , was used in this study, which was deposited and maintained at BIOTEC Culture Collection (BCC), Pathumthani, Thailand as strain BCC 15763 T and at Culture Collection NBRC, Kisarazu, Japan as strain NBRC 101099 T . The strain was isolated in September 2002 from a flower of red ginger (khing daeng in Thai; Alpinia purpurata) collected in Chiang Mai, Thailand, at pH 3.5 by the enrichment culture approach using a glucose/ethanol/acetic acid (0.3%, w/v) medium, but not either a sorbitol medium, a dulcitol medium, or a sucrose medium (Katsura et al, 2001;Lisdiyanti et al, 2002;Yamada et al, 1976Yamada et al, , 1999Yamada et al, , 2000Yukphan et al, 2004c T was sequenced for 16S rRNA genes, as described previously (Yukphan et al, 2004a, b). A gene fragment specific for 16S rRNA gene-coding regions was amplified by means of a PCR amplification.…”

“…The genus was quite differentiated in these respects from the genera Kozakia and Swaminathania, which showed no motility, the ability to oxidize ethanol to acetic acid, and growth in the presence of 0.35% acetic acid (w/v) (Katsura et al, 2001;Lisdiyanti et al, 2002;Loganathan and Nair, 2004;Yamada et al, 2000;Yukphan et al, 2004c).…”

“…Morphological, physiological, and biochemical characteristics were tested by the methods of Asai et al (1964), Yamada et al (1976Yamada et al ( , 1999Yamada et al ( , 2000, Katsura et al (2001), and Yukphan et al (2004c).…”

“…Recently, the additional two species, Asaia siamensis Katsura et al 2001 and Asaia krungthepensis Yukphan et al 2004 were described (Katsura et al, 2001;Yukphan et al, 2004c). The genus Kozakia Lisdiyanti et al 2002 was subsequently proposed as the sixth genus with a single species, Kozakia baliensis Lisdiyanti et al 2002(Lisdiyanti et al, 2002.…”

Gluconobacter wancherniae sp. nov., an acetic acid bacterium from Thai isolates in the α-Proteobacteria

“…Two isolates, designated as AC42 T (= BCC 15775 T = NBRC 103581 T ) and AC47 (= BCC 15780 = NBRC 103582), were isolated respectively from an unknown seed and an unknown yellow fruit by an enrichment culture approach using glucose/ethanol/acetic acid medium (Yamada et al, 1976(Yamada et al, , 1999(Yamada et al, , 2000Yukphan et al, 2004c), and examined phylogenetically, genetically, phenotypically, and chemotaxonomically. Extraction and isolation of bacterial chromosomal DNAs were performed by a modifi cation of the methods of Marmur (1961) (Ezaki et al, 1983;Saito and Miura, 1963).…”