Characterization of haloglycan, an exopolysaccharide produced by Halomonas stenophila HK30.

We have conducted a thorough study of the exopolysaccharide (EPS) produced by strain HK30 of Halomonas stenophila, which we have named haloglycan. This strain was chosen during an ongoing research programme aimed at finding novel exopolysaccharide-producing halophilic bacteria in unexplored hypersaline habitats. Strain HK30 was isolated from a saline-wetland in Brikcha (Morocco) and identified as belonging to the species H. stenophila. It produced EPS mainly during the exponential growth phase and to a lesser extent during the stationary phase. Culture parameters influenced both bacterial growth and EPS production, EPS yield always being directly related to the quantity of biomass. Under optimum culture conditions, strain HK30 produced 3.89 g of EPS per litre of medium. The polymer was a sulphated heteropolysaccharide composed of two fractions, with molecular masses of 8.2 × 10(4) and 1.4 × 10(6). The crude EPS contained 44 ± 0.1% w/w carbohydrates and the following monosaccharide composition: glucose (24 ± 1.73), glucuronic acid (7.5 ± 0.37), mannose (5.5 ± 0.17), fucose (4.5 ± 0.36), galactose (1.2 ± 0.17) and rhamnose (1 ± 0.05) (%, w/w). It produced solutions of high viscosity and pseudoplastic behaviour that showed interesting flocculating and emulsifying activities and was also involved in forming biofilm.

The potential biotechnological applications of the exopolysaccharide produced by the halophilic bacterium Halomonas almeriensis.

We have studied the extracellular polysaccharide (EPS) produced by the type strain, M8(T), of the halophilic bacterium Halomonas almeriensis, to ascertain whether it might have any biotechnological applications. All the cultural parameters tested influenced both bacterial growth and polysaccharide production. EPS production was mainly growth-associated and under optimum environmental and nutritional conditions M8(T) excreted about 1.7 g of EPS per litre of culture medium (about 0.4 g of EPS per gram of dry cell weight). Analysis by anion-exchange chromatography and high-performance size-exclusion chromatography indicated that the exopolysaccharide was composed of two fractions, one of 6.3 × 10(6) and another of 1.5 × 10(4) Daltons. The monosaccharide composition of the high-molecular-weight fraction was mannose (72% w/w), glucose (27.5% w/w) and rhamnose (0.5% w/w). The low-molecular-weight fraction contained mannose (70% w/w) and glucose (30% w/w). The EPS has a substantial protein fraction (1.1% w/w) and was capable of emulsifying several hydrophobic substrates, a capacity presumably related to its protein content. The EPS produced solutions of low viscosity with pseudoplastic behaviour. It also had a high capacity for binding some cations. It contained considerable quantities of sulphates (1.4% w/w), an unusual feature in bacterial polysaccharides. All these characteristics render it potentially useful as a biological agent, bio-detoxifier and emulsifier.

Halomonas rifensis sp. nov., an exopolysaccharide-producing, halophilic bacterium isolated from a solar saltern.

A polyphasic taxonomic study was conducted on strain HK31(T), a moderately halophilic bacterium isolated from a solar saltern in Chefchaouen, Morocco. The strain was a Gram-reaction-negative, oxidase-positive rod, which was motile by means of peritrichous flagella. The strain required NaCl for growth and grew in salt concentrations (mixture of sea salts) of 0.5-20 % (w/v) (optimum 5-7.5 %, w/v), at 25-45 °C (optimum 32 °C) and at pH 5-10 (optimum pH 6-9). Strain HK31(T) did not produce acids from sugars and its metabolism was respiratory, using oxygen as terminal electron acceptor. The strain was positive for the accumulation of poly-ß-hydroxyalkanoate granules and formed mucoid colonies due to the excretion of an exopolysaccharide. The DNA G+C content was 61.5 mol%. 16S rRNA gene sequence analysis indicated that it belonged to the genus Halomonas in the class Gammaproteobacteria. The most phylogenetically related species was Halomonas anticariensis, with which strain HK31(T) showed a 16S rRNA gene sequence similarity of 96.48 %. Its major fatty acids were C(18 : 1)ω7c, C(16 : 0), C(19 : 0) cyclo ω8c, C(16 : 1)ω7c/iso-C(15 : 0) 2-OH and C(12 : 0) 3-OH and the predominant respiratory lipoquinone was ubiquinone with nine isoprene units (Q-9). Based on the evidence provided in this study, strain HK31(T) (= CECT 7698(T) = LMG 25695(T)) represents a novel species of the genus Halomonas, for which the name Halomonas rifensis is proposed.