Gracilibacillus alcaliphilus sp. nov., a facultative alkaliphile isolated from indigo fermentation liquor for dyeing

Abstract

A facultatively alkaliphilic, lactic-acid-producing and halophilic strain, designated SG103 T , was isolated from a fermented Polygonum indigo (Polygonum tinctorium Lour.) liquor sample for dyeing prepared in a laboratory. 16S rRNA gene sequence phylogeny suggested that SG103T is a member of the genus Gracilibacillus with the closest relatives being 'Gracilibacillus xinjiangensis' J2 (similarity: 97.06 %), Gracilibacillus thailandensis TP2-8 T (97.06 %) andGracilibacillus halotolerans NN T (96.87 %). Cells of the isolate stained Gram-positive and were facultatively anaerobic straight rods that were motile by peritrichous flagella. The strain grew at temperatures between 13 and 48 6C with the optimum at 39 6C. It grew in the range pH 7-10 with the optimum at pH 9. The isoprenoid quinone detected was menaquinone-7 (MK-7) and the DNA G+C content was 41.3 mol%. The whole-cell fatty acid profile mainly (.10 %) consisted of iso-C 15 : 0 , anteiso-C 15 : 0 and anteiso-C 17 : 0 . Unlike other reported species of the genus Gracilibacillus, the strain lacked diphosphatidylglycerol as a major polar lipid. DNA-DNA hybridization experiments with strains exhibiting greater than 96.87 % 16S rRNA gene sequence similarity, 'G. xinjiangensis' J2, G. thailandensis TP2-8 T and G. halotolerans NN T , revealed 2±4 %, 4±9 % and 3±2 % relatedness, respectively. On the basis of the differences in phenotypic and chemotaxonomic characteristics, and the results of phylogenetic analyses based on 16S rRNA gene sequences and DNA-DNA relatedness data from reported species of the genus Gracilibacillus, strain SG103 T merits classification as a members of a novel species, for which the name Gracilibacillus alcaliphilus sp. nov. is proposed. The type strain is SG103 T (5JCM 17253 T 5NCIMB 14683 T ).During indigo fermentation, which proceeds under alkaline conditions (higher than pH 10), the pH gradually decreases owing to the action of micro-organisms (Aino et al., 2010). We have isolated indigo-reducing and lactic-acid-producing bacteria, namely Alkalibacterium psychrotolerans, Alkalibacterium iburiense and Alkalibacterium indicireducens, from indigo fermentation liquor obtained from Date City, Iburi, Hokkaido and Tokushima City, Shikoku in Japan (Nakajima et al., 2005;Yumoto et al., 2004Yumoto et al., , 2008. A lacticacid-producing bacterium, Oceanobacillus polygoni, has also been isolated from a fermented indigo liquor sample prepared in a laboratory (Hirota et al., 2013). In the present study, we further screened for new lactic-acid-producing micro-organisms from indigo fermentation liquor and found that one strain belonging to the genus Gracilibacillus produced lactic acid.The genus Gracilibacillus is one of the moderate halophilic genera belonging to the family Bacillaceae. Since the first description of the genus Gracilibacillus with Gracilibacillus halotolerans, which was isolated from surface mud of the Great Salt Lake in the USA (Wainø et al., 1999), and Bacillus dipsosauri (Lawson et al., 1996) from the salt Abbreviation: meso-DAP, mes...

Genome sequence and description of Gracilibacillus timonensis sp. nov. strain Marseille‐P2481T, a moderate halophilic bacterium isolated from the human gut microflora

“… G. timonensis strain Marseille‐P2481 T , G. Gracilibacillus bigeumensis strain BH097 T (Kim et al., ) , G. halophilus strain YIM‐C55.5 T (Chen et al., ) , G. boraciitolerans strain T‐16X T (Ahmed et al., ) , G. saliphilus strain YIM91119 T (Tang et al., ) , G. kekensis strain K170 T (Gao et al., ) , G. halotolerans strain NN T (Wainø et al., ) , G. alcaliphilus strain SG103 T (Hirota, Hanaoka, Nodasaka, & Yumoto, ). …”

“…Classification and general features of Gracilibacillus timonensis strain Marseille-P2481 T according to the MIGS recommendations [23] TA B L E 2 Differential characteristics of Gracilibacillus timonensis strain Marseille-P2481 T and other closely related members of the genus Gracilibacillus Properties data available. G. timonensis strain Marseille-P2481 T , G. Gracilibacillus bigeumensis strain BH097 T (Kim et al, 2012), G. halophilus strain YIM-C55.5 T (Chen et al, 2008), G. boraciitolerans strain T-16X T (Ahmed et al, 2007),G. saliphilus strain YIM91119 T(Tang et al, 2009), G. kekensis strain K170 T(Gao et al, 2012), G. halotolerans strain NN T(Wainø et al, 1999), G. alcaliphilus strain SG103 T(Hirota, Hanaoka, Nodasaka, & Yumoto, 2014).…”

Microbial culturomics unravels the halophilic microbiota repertoire of table salt: description of Gracilibacillus massiliensis sp. nov.

“… G. massiliensis Awa-1 T ; G. thailandensis TP2-8 T ( 9 ); G. orientalis XH-63 T ( 39 ); G. ureilyticus MF38 T ( 6 ); G. halophilus YIM-C55.5 T ( 8 ); G. boraciitolerans T-16X T ( 40 ); G. saliphilus YIM91119 T ( 41 ); G. kekensis K170 T ( 11 ); G. halotolerans NN T ( 5 ); G. alcaliphilus SG103 T ( 7 ). NA=not available.…”

“…It was selected for sequencing based on its phenotypic differences, phylogenetic position, and 16S rRNA sequence similarity with other members of the genus Gracilibacillus . The G+C content of the genomic DNA varies from 35.3 to 42.3 mol% ( 7 ). According to the fact that the G+C content deviation within species is at most 1%, these values confirm the classification of strain Awa-1 T in a distinct species ( 42 ).…”

“…First described by Wainø et al in 1999 ( 5 ), the genus Gracilibacillus includes, moderately halophilic or halotolerant, mobile, gram-positive bacteria, most of them forming endospores or filaments containing menaquinone-7 (MK-7) as predominant respiratory quinone ( 6 ). This genus includes 12 species ( www.bacterio.net ) described with valid published names ( 7 ). Members of the genus Gracilibacillus are salty environmental bacteria isolated most often from soil ( 8 ), food ( 9 ), lakes and salty sea water ( 10 , 11 ).…”