Abstract
The halophilic bacteria Halomonas eurihalina produces an exopolysaccharide (EPS) of potential interest for biotechnological applications. To study the characteristics of this bacteria and others like it, we have developed an efficient chemical mutation technique using hydroxylamine and, in this way, have obtained non-mucoid mutants of its F2-7 strain. We have characterised the phenotypes of the mutants via nutritional, biochemical and physiological tests and by their susceptibility to antimicrobial agents and heavy metals. These mutants will be used as recipient tools in further studies to discover the genetic determinants for the synthesis of EPS.
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References
Béjar V, Llamas I, Calvo C, Quesada, E (1998) Characterization of exopolysaccharides produced by 19 halophilic strains of the species Halomonas eurihalina. J. Biotechnol. 61: 135-141.
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-Dye binding. Anal. Biochem. 72: 248-254.
Calvo C, Ferrer MR, Martínez-Checa F, Béjar V, Quesada E (1995) Some rheological properties of the extracellular polysaccharide produced by Volcaniella eurihalina F2-7. Appl. Biochem. Biotech. 55: 45-54.
Calvo C, Martínez-Checa F, Mota F, Béjar V, Quesada E (1998) Effect of cations, pH and sulphate content on the viscosity and emulsifying activity of the Halomonas eurihalina exopolysaccharide. J. Indust. Microbiol. Biotechnol. 20: 205-209.
Cánovas D, Vargas C, Ventosa A, Nieto JJ (1997) Salt-sensitive and auxotrophic mutants of Halomonas elongata and H. meridiana by use of hidroxylamine mutagenesis. Curr. Microbiol. 34: 85-90.
Dubois M, Gilles KA, Hamilton JK, Rebers PA, Smith F (1956) Colorimetric method for determination of sugars and related substances. Anal. Chem. 28: 350-356.
Fernández-Castillo R, Nieto JJ, Megías M, Ruíz-Berraquero F (1990) Efficient hydroxylamine mutagenesis of Haloferax mediterranei and other halophilic archaebacteria. Curr. Microbiol. 21: 83-89.
Llamas I, del Moral A, Girón MD, Salto R, Béjar V, Quesada E (1997) Plasmids from Halomonas eurihalina, a microorganism which produces an exopolysaccharide of biotecnological interest. FEMS Microbiol. Lett. 156: 251-257.
Mellado E, Nieto JJ, Ventosa A (1995) Phylogenetic interferences and taxonomic consequences of 16S ribosomal DNA sequence comparison of Chromohalobacter marismortui, Volcaniella eurihalina and Deleya halophila and reclassification of V. eurihalina as Halomonas eurihalina comb. nov. Int. J. Syst. Bacterial. 45: 712-716.
Moraine RA, Rogovin P (1966) Kinetics of polysaccharide B-1459 fermentation. Biotech. Bioeng. 8: 511-524.
Morelle G (1989) A plasmid extraction procedure on a miniprep scale. BRL Focus 11: 7-8.
Nieto JJ, Fernández-Castillo R, Márquez MC, Ventosa A, Quesada E, Ruíz-Berraquero F (1989) A survey of metal tolerance in moderately halophilic eubacteria. Appl. Environ. Microbiol. 55: 2385-2390.
Nieto JJ, Fernández-Castillo R, Megías M, Ruíz-Berraquero F (1992) Ethyl methanesulfonate mutagenesis in extremely halophilic archaebacteria: isolation of auxotrophic mutants of Haloferax mediterranei and Haloferax gibbonsii. Curr. Microbiol. 24: 41-47.
Quesada E, Béjar V, Valderrana MJ, Ramos-Cormenzana A (1987) Growth characteristics and salt requirement of Deleya halophila in a defined medium. Curr. Microbiol. 16: 21-25.
Quesada E, Valderrama MJ, Béjar V, Ventosa A, Gutiérrez MC, Ruíz-Berraquero F, Ramos-Cormenzana, A (1990)Volcaniella eurihalina gen nov, sp nov, a moderately halophilic non motile gram-negative rod. Int. J. Syst. Bacterial. 40: 261-267.
Quesada E, Béjar V, Calvo C (1993) Exopolysaccharide production by Volcaniella eurihalina. Experientia 49: 1037-1041.
Quesada E, del Moral A, Béjar, V (1994) Comparative methods for isolation of Volcaniella eurihalina exopolysaccharide. Biotech. Tech. 8: 701-706.
Rodríguez-Valera F, Ruíz-Berraquero F, Ramos-Cormenzana, A (1981) Characteristics of heterotrophic bacterial populations in hypersaline environments of different salt concentrations.Microbial. Ecol. 7: 235-243.
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular Cloning: A Laboratory Manual, 2nd edn. New York: Cold Spring Harbor Laboratory Press.
Sutherland IW (1990)Cambridge Studies in Biotechnology. Biotechnology of Microbial Exopolysaccharides, Vol. 9. Cambridge: Cambridge University Press.
Sutherland IW (1998) Novel and established applications of microbial polysaccharides. Trends. Biotechnol. 16: 41-46.
Thorne L, Tansey L, Pollock TJ (1987) Clustering of mutation blocking synthesis of xanthan gum by Xanthomonas campestris. J. Bacterial. 169: 3593-3600.
Tombs MP, Harding SE (1998) An Introduction to Polysaccharide Biotechnology, London: Taylor and Francis.
Ventosa A, Nieto J, Oren A (1998) Biology of moderately halophilic aerobic bacteria. Microbiol. Mol. Biol. Rev. 62: 504-544.
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Llamas, I., Béjar, V., Argandoña, M. et al. Chemical mutagenesis of Halomonas eurihalina and selection of exopolysaccharide-deficient variants. Biotechnology Letters 21, 367–370 (1999). https://doi.org/10.1023/A:1005483004140
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DOI: https://doi.org/10.1023/A:1005483004140