Alkaliphilus crotonatoxidans sp. nov., a strictly anaerobic, crotonate-dismutating bacterium isolated from a methanogenic environment

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Abstract
Two bacterial strains were isolated from methanogenic butyrate-oxidizing mixed cultures. The cells were straight to slightly curved, Gram-positive rods that were motile by means of multiple flagella and formed endospores. Growth was observed in the temperature range 15–45 °C (optimum 37 °C) and pH range 5·5–9·0 (optimum pH 7·5). The novel isolates were strictly anaerobic chemo-organotrophs capable of utilizing yeast extract, peptone, tryptone and a variety of sugars and organic acids, but not glucose. None of the accessory electron acceptors tested (elemental sulfur, thiosulfate or fumarate) improved growth, except crotonate, which was dismutated to butyrate and acetate. The G+C content of the DNA of one of the isolates, strain B11-2T, was 30·6 mol%. Phylogenetic analysis based on 16S rDNA sequence similarity between strain B11-2T and some other strictly anaerobic, spore-forming bacteria indicated that the novel isolates represented a species in cluster XI within the low-GC Gram-positive bacteria, being most closely related to Alkaliphilus transvaalensis JCM 10712T. DNA–DNA relatedness between strain B11-2T and A. transvaalensis JCM 10712T was 21 %. On the basis of physiological and molecular properties, and cellular fatty acid and cell wall compositions, the novel isolates are proposed to represent a novel species of the genus Alkaliphilus, for which the name Alkaliphilus crotonatoxidans is proposed (type strain B11-2T=AS 1.2897T=JCM 11672T).

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