Classifications

• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12H—PASTEURISATION, STERILISATION, PRESERVATION, PURIFICATION, CLARIFICATION OR AGEING OF ALCOHOLIC BEVERAGES; METHODS FOR ALTERING THE ALCOHOL CONTENT OF FERMENTED SOLUTIONS OR ALCOHOLIC BEVERAGES
  • C12H1/00—Pasteurisation, sterilisation, preservation, purification, clarification, or ageing of alcoholic beverages
  • C12H1/02—Pasteurisation, sterilisation, preservation, purification, clarification, or ageing of alcoholic beverages combined with removal of precipitate or added materials, e.g. adsorption material
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
  • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
  • A23L29/20—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents
  • A23L29/269—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents of microbial origin, e.g. xanthan or dextran
• • C—CHEMISTRY; METALLURGY
  • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
  • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
  • C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
  • C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof

Definitions

• the present invention relates to a new polysaccharide produced by a marine bacterium.
• the bacterial subspecies Alteromonas macleodii subsp. fij iensis has been isolated from underwater hydrothermal vents. When cultivated under laboratory conditions on a medium rich in glucose, this bacterium secretes exopolysaccharides in large quantities.
• the morphological, physiological and phylogenetic characteristics of Alteromonas ma cleodii subsp. fijiensis are indicated in the publication of RAGUENES et al., [Applied and Environmental Microbiology, 62 (1), pp. 67-73, (1996)].
• This publication also gives indications on the overall composition of the polysaccharide mixture produced by a strain of Al teromonas macleodii subsp. fijiensis called ST716 cultivated on medium enriched in glucose, and recovered from this medium by precipitation with ethanol.
• This polysaccharide mixture comprises (percentages by mass) 10.5% glucose, 11.1% galactose, 4.4% mannose, 5% mannose pyruvate, 12.2% glucuronic acid and 6.3% d galacturonic acid, corresponding to respective molar ratios of glucose, galactose, mannose, glucuronic acid, and galacturonic acid of 1.0 / 1.1 / 0.6 / 1.1 / 0.6. Its sulphate content is 5%.
• the inventors sought to identify the constituents of this mixture, and purified from it a new polysaccharide.
• This purified polysaccharide which is the subject of the present invention, consists of glucose, galactose, glucuronic acid, galacturonic acid, and mannose pyruvate. It is characterized in that these various constituents are respectively represented in the molar ratios 1/1/1/2/1, and associated in a repeating hexasaccharide unit in which three osidic residues form a main chain, the branching point of which is constituted a residue of galacturonic acid. On the latter is grafted a side chain ending in a residue of mannose pyruvate in position 4 and in position 6.
• a polysaccharide according to the invention consists of n hexasaccharide units (n being equal to or greater than 1) corresponding to the formula (I) below: -> 4) - ⁇ -D-Glcp- (1 ⁇ 4) - ⁇ -D-GalpA- (1 ⁇ 4) - ⁇ -D-Galp- (l ⁇
• a polysaccharide in accordance with the invention can, for example, be purified from the exopolysaccharides produced by the strain ST716.
• This strain was deposited by IFREMER, according to the Budapest Treaty, on October 17, 1995 with the CNCM (National Collection of Cultures of Microorganisms) held by the Institut Pasteur, 25 rue du Dondel Roux, in Paris, under number 1 -1627.
• a polysaccharide of formula (I), purified by anion exchange chromatography from the mixture of exopolysaccharides secreted by Al teromonas macleodii subsp. Fij iensis, has an average molecular weight (M w ) of the order of 10 6 Da, or an average degree of polymerization DP n of the order of 800.
• the invention therefore also relates to the use of a polysaccharide according to the invention as a thickening agent, as well as as a shear-thinning agent.
• a polysaccharide according to the invention as a thickening agent, as well as as a shear-thinning agent.
• the invention also relates to the use of a polysaccharide according to the invention as a texturing agent, in particular stabilizer and / or thickener in the food industry. It relates more particularly to the use of a polysaccharide according to the invention for the preparation of a product intended for human or animal consumption.
• the polysaccharides according to the invention can, for example, be used to impart the desired consistency to liquid, creamy products, pasty, gelled or semi-solid, while improving and / or stabilizing their texture.
• They can also be used as a moisture retainer in solid products, to give them and keep them a soft texture.
• polysaccharides in accordance with the invention are very easily and homogeneously incorporated into all the preparations carried out in the presence of water. Their use in the context of industrial processes is therefore very simple, and, moreover, they are particularly suitable for the manufacture of “ready-to-use” products to be reconstituted by the end consumer.
• the polysaccharides according to the invention can also be used for the clarification of liquid products for food use, such as wines or vinegar.
• the present invention also encompasses products intended for human or animal consumption, comprising at least one polysaccharide according to the invention.
• the polysaccharides in accordance with the invention are advantageously present in the products in question at a rate of 0.01 to 10%, preferably 0.05 to 5% by weight, relative to the total weight of the ingredients.
• exopolysaccharides secreted by the strain of Al teromonas macleodii subsp. fij iensis C.N.C.M 1-1627 are obtained from cultures of this bacterium by ethanol precipitation, as described by RAGUENES et al., [Applied and Environmental Microbiology, 62 (1), pp. 67-73, (1996)].
• the elution profile is shown in Figure 1.
• the main peak fractions corresponding to an NaCl concentration of 450 mM were pooled, dialyzed and lyophilized.
• composition The polysaccharide obtained as described in Example 1 is subjected to acidic methanolysis and the methylglycosides are converted into corresponding trimethylsilylated derivatives according to the method of MONTREUIL et al. [Carbohydrate analysis: a practical approach, Chaplin M. F. and Kennedy J.K. (Eds), I.R.L. Press Oxford, Washington D.C., Glycoproteins pp.143-204 (1986)], then analyzed by gas chromatography coupled to a mass spectrometer.
• Neutral dares are also analyzed in the form of alditol acetates after hydrolysis of the polysaccharide, followed by reduction and acetylation according to the method of BLAKENEY et al. [Carbohydr. Res., 113, pp. 291-299, (1983)].
• the pyruvic acid content is estimated using a colorimetric assay method using 2,4-dinitrophenylhydrazone [SLONEKER et al., Nature, 194, pp. 478-479, (1962)].
• the alditol acetates resulting from the reduction then from the acetylation of the partially methylated residues are analyzed by coupling gas chromatography / mass spectrometry. Their nature, characterized by the position of their methyl groups, defines the cyclic form as well as the position of the glycosidic bonds of the corresponding residues.
• This analysis shows the presence of glucose and galactose substituted at 4, mannose di-substituted at 4 and 6 (confirming the substitution sites of the pyruvate group), glucuronic acid substituted at 3, glucuronic acid substituted at 4 and of galacturonic acid di-substituted in 3 and in 4.
• the latter therefore constitutes a connection point of a repeating branched hexasaccharide unit whose end of the side chain is the residue of mannose pyruvate. All residues are cyclic pyranose. Location of uronic acids
• the previously methylated polysaccharide is treated in basic medium, according to the method of
• the repeating unit therefore comprises a sequence of three uronic acid residues, linked in position 4 of the galactose residue.
• the NMR spectra were recorded at 320K on BRUKER AMX-500 devices and AMX-600.
• the high field analyzes in homonuclear mode "TOCSY” (Total Observed Correlation SpectroscopY) and "COZY” (Correlation Observed SpectroscopY) as well as the spectrum obtained in high field in heteronuclear mode "HSQC” (Heteronuclear Single Quantum Coherence) assignment of signals to the protons and carbons of each residue.
• TOCSY Total Observed Correlation SpectroscopY
• COZY Correlation Observed SpectroscopY
• the ⁇ / ⁇ configuration of the glycosidic bonds is defined by the value of the chemical shifts of the signals corresponding to the anomeric carbons and protons.
• Analysis in homonuclear mode "NOESY” Nuclear Overhauser Enhancement SpectroscopY determines the sequence of residues of the repeating unit.
• EXAMPLE 3 DETERMINATION OF THE RHEOLOGICAL PROPERTIES OF POLYSACCHARIDE.
• the shape of the flow curve demonstrates a shear thinning behavior, with the existence of a so-called “Newtonian” plateau in the field of low values of shear speed followed by a decrease in the apparent viscosity as a function shear speed for high speed values.
• EXAMPLE 4 USE OF A POLYSACCHARIDE CONFORMING TO THE INVENTION FOR THE PREPARATION OF FOODSTUFFS

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• Chemical & Material Sciences (AREA)
• Engineering & Computer Science (AREA)
• Health & Medical Sciences (AREA)
• Life Sciences & Earth Sciences (AREA)
• Organic Chemistry (AREA)
• Biochemistry (AREA)
• Food Science & Technology (AREA)
• Polymers & Plastics (AREA)
• General Health & Medical Sciences (AREA)
• Dispersion Chemistry (AREA)
• General Engineering & Computer Science (AREA)
• Materials Engineering (AREA)
• Chemical Kinetics & Catalysis (AREA)
• Medicinal Chemistry (AREA)
• Molecular Biology (AREA)
• Bioinformatics & Cheminformatics (AREA)
• Nutrition Science (AREA)
• Genetics & Genomics (AREA)
• Wood Science & Technology (AREA)
• Zoology (AREA)
• Polysaccharides And Polysaccharide Derivatives (AREA)
• Jellies, Jams, And Syrups (AREA)
• Preparation Of Compounds By Using Micro-Organisms (AREA)

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Citations (4)

• 1998
  • 1998-06-22 FR FR9807839A patent/FR2780063B1/fr not_active Expired - Fee Related
• 1999
  • 1999-06-22 EP EP99957183A patent/EP1171625B1/fr not_active Expired - Lifetime
  • 1999-06-22 US US09/720,238 patent/US6545145B1/en not_active Expired - Fee Related
  • 1999-06-22 AU AU42704/99A patent/AU4270499A/en not_active Abandoned
  • 1999-06-22 JP JP2000556051A patent/JP2002518586A/ja not_active Ceased
  • 1999-06-22 DE DE69923356T patent/DE69923356T2/de not_active Expired - Fee Related
  • 1999-06-22 WO PCT/FR1999/001490 patent/WO1999067411A1/fr not_active Ceased

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