EP2616547A1 - Procédé de production de l-fucose - Google Patents

Procédé de production de l-fucose

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Publication number
EP2616547A1
EP2616547A1 EP11769803.5A EP11769803A EP2616547A1 EP 2616547 A1 EP2616547 A1 EP 2616547A1 EP 11769803 A EP11769803 A EP 11769803A EP 2616547 A1 EP2616547 A1 EP 2616547A1
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EP
European Patent Office
Prior art keywords
fucose
fermentation
glucose
polysaccharide
ppm
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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Application number
EP11769803.5A
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German (de)
English (en)
Inventor
Alessandro Gori
Silvia Biagiolini
Marco Manoni
Luana Vagnoli
Liana Salsini
Jacopo Chini
Silvia Giacomelli
Giovanni Cipolletti
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Inalco SpA
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Inalco SpA
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Publication date
Application filed by Inalco SpA filed Critical Inalco SpA
Publication of EP2616547A1 publication Critical patent/EP2616547A1/fr
Withdrawn legal-status Critical Current

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Classifications

    • 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
    • C08B37/0063—Glycosaminoglycans or mucopolysaccharides, e.g. keratan sulfate; Derivatives thereof, e.g. fucoidan
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
    • C—CHEMISTRY; METALLURGY
    • C13—SUGAR INDUSTRY
    • C13K—SACCHARIDES OBTAINED FROM NATURAL SOURCES OR BY HYDROLYSIS OF NATURALLY OCCURRING DISACCHARIDES, OLIGOSACCHARIDES OR POLYSACCHARIDES
    • C13K13/00—Sugars not otherwise provided for in this class
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales

Definitions

  • the present invention relates to a process for the production, on an industrial scale, of L-fucose (or 6-deoxygalactose) by hydrolysis of a polysaccharide produced by a fermentation process effected by a novel isolated microbial strain belonging to the family Enterobacteriaceae.
  • Fucosylated oligosaccharides and polysaccharides are of great interest to the chemical, pharmaceutical, cosmetic and nutraceutical industry. Fucosylated oligosaccharides and polysaccharides are known to have potential medical applications as anti-tumour and anti-inflammatory agents but also potential cosmetic applications as anti-ageing agent for the skin that is able to improve skin tonicity.
  • Fucosyl derivatives for which L-fucose represents the raw material, are of industrial interest for their known antiallergic, hydrating, emulsifying and stabilizing properties.
  • Another very important use of L-fucose is as a precursor of semisynthetic analogues of the fucosylated oligosaccharides contained in human milk (human milk oligosaccharides).
  • the production of L-fucose industrially is still rather problematic both with respect to chemical and biochemical synthesis, and conversely with respect to the use of bacterial strains that produce L-fucose or polysaccharides containing L-fucose by fermentation.
  • EP0102535 describes the fermentation of bacteria of the genera Alcaligenes, Klebsiella, Pseudomonas or Enterobacter for producing extracellular polysaccharides rich in rhamnose or fucose. These deoxy-sugars can, after acid hydrolysis of the polysaccharide, be isolated from the hydrolysate.
  • the L-fucose currently on the market is obtained from natural sources, in particular by acid hydrolysis of sulphated polysaccharides such as fucoidans extracted from algae such as Laminaria.
  • sulphated polysaccharides such as fucoidans extracted from algae such as Laminaria.
  • direct extraction from algae is expensive and is moreover subject to seasonal variations in volumes and product quality.
  • L-fucose Another source of L-fucose is the bark of trees with a high trunk such as willow, birch and beech. In this case too, L-fucose is extracted by hydrolysis, with low yields and high costs.
  • the present invention solves the aforementioned problems by means of a new bacterial strain belonging to the family Enterobacteriaceae, identified as C2B2, isolated from foliar extract of Phalaenopsis sp.
  • Fig.1 shows the plot of a 1 H-NMR spectrum (in deuterated water, performed at 25°C with acetonitrile internal standard at 2.06ppm) of a fucose-containing polysaccharide, produced by a bacterial strain according to the invention (as from example 6)
  • Fig.2 shows the plot of a 13 C-NMR spectrum (in deuterated water, performed at 25°C with acetonitrile internal standard at 1 .470ppm) of a fucose-containing polysaccharide, produced by a bacterial strain according to the invention (as from example 6)
  • Fig.3 shows the plot of a 13 C-NMR spectrum (in deuterated water, performed at 25°C with acetonitrile internal standard at 1 .470ppm) of the anomeric region of a fucose- containing polysaccharide, produced by a bacterial strain according to the invention (as from example 6)
  • Fig.4 shows an HPLC plot of a partial hydrolysis mixture of a polysaccharide according to the invention (as from example 7)
  • Fig.5 shows an HPLC plot of a partial hydrolysis mixture of a polysaccharide according to the invention, which was then treated with ion exchange resins to eliminate the peak at about 1 1 minutes (as from example 7).
  • Fig.6 shows an HPLC plot of a complete hydrolysis mixture of a polysaccharide according to the invention (as from example 8)
  • Fig.7 shows an HPLC plot of a partial hydrolysis mixture of a polysaccharide according to the invention (as from example 9)
  • the C2B2 bacterial strain was isolated from foliar extract of orchids of the genus Phalaenopsis.
  • the new strain isolated was identified as belonging to the family Enterobacteriaceae and was designated C2B2.
  • the new strain was deposited at the Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSMZ) (German Collection of Microorganisms and Cell Cultures), Inhoffenstr. 7 B, D-38124, Braunschweig (Germany) on 23 January 2009 in accordance with the provisions of the Budapest Treaty.
  • DSMZ Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH
  • the new microbial strain was assigned the accession number DSM 22227.
  • the microbial strain according to the invention was characterized by sequencing the ribosomal RNA 16S resulting in a SEQ ID NO: 1 given below:
  • the present invention also relates to microbial strains belonging to the family Enterobacteriaceae, comprising a sequence rRNA 16S containing a sequence with homology greater than or equal to 93% with SEQ. ID NO: 1 ; preferably said homology is greater than or equal to 98% with SEQ. ID NO: 1 .
  • the present invention also relates to mutated strains derived from the C2B2 strain.
  • These strains can be obtained by simple selection of strains spontaneously mutated and isolated from a culture of C2B2 or by selection of strains derived from C2B2 by the action of mutagenic factors, such as UV radiation or X-rays, or by the action of chemicals such as ozone, nitrous acid, N-methyl-N'-nitro-N-nitrosoguanidine (NTG) or ethane methane sulphonate (EMS).
  • mutagenic factors such as UV radiation or X-rays
  • chemicals such as ozone, nitrous acid, N-methyl-N'-nitro-N-nitrosoguanidine (NTG) or ethane methane sulphonate (EMS).
  • said derived or mutated microbial strain comprises an rRNA 16S sequence containing a sequence having homology with SEQ. ID NO: 1 equal or greater than 98%.
  • the C2B2 bacterial strain according to the invention has the following morphological and biochemical characteristics:
  • the present invention also relates to the fermentation process developed for the production of extracellular polysaccharides containing L-fucose and the subsequent isolation of 6-deoxy sugar.
  • the C2B2 strain is grown by inoculation in any fermentable aqueous medium containing carbon and nitrogen sources and mineral salts.
  • the source of assimilable carbon is represented by one or more sugars (for example glucose, fructose, maltose, sucrose, starch, mannitol, sorbitol, lactose, corn syrup etc.) alone or mixed.
  • sugars for example glucose, fructose, maltose, sucrose, starch, mannitol, sorbitol, lactose, corn syrup etc.
  • glucose is a preferred carbon source.
  • the nitrogen source used can be proteinaceous material of various types (for example yeast extract, soya flour, hydrolysed proteins, corn steep liquor etc.). Typically the nitrogen source is between 0.05 and 0.5 wt.% in the medium.
  • Various salts that are normally used in nutrient media can be used.
  • Non-limiting examples are: phosphates, sulphates, chlorides, sodium carbonate, potassium, ammonium, calcium and magnesium.
  • the fermentation can be carried out at temperatures between 25 and 35 °C, preferably 30°C, at a pH between 6.0 and 7.5 maintained by means of systems for automatic correction, with aeration and agitation, for a period of 2-5 days.
  • L-fucose The formation of L-fucose is monitored by HPLC on samples submitted beforehand to complete acid hydrolysis; macroscopically it is possible to assess the progress of fermentation on the basis of the viscosity, which, in a medium where the expected quantity of polysaccharide is being formed, reaches and even exceeds 2200 cP as described in the literature (P.T. Vanhooren et al. Med. Fac. Landbouww. Univ. Gent., vol. 62/4a, p. 1271 -1276, 1997).
  • Fermentation can be carried out in conventional fermenters by inoculating the nutrient medium with a culture of the C2B2 strain. Prior to inoculation the nutrient medium is sterilized, for example by heat at temperatures of the order of 120°C.
  • the culture suspension is submitted to suitable treatments for the purpose of purification and recovery of L-fucose.
  • NMR spectra (not shown) acquired for example at a temperature of 40°C also show anomeric signals at about 4.7ppm and about 4.6ppm in approx. 1 :1 ratio and attributable to bonds of the beta type.
  • the polysaccharides obtained from fermentation of the C2B2 strain preferably have, moreover, 1 H-NMR signals at about 1 .4ppm (broad singlet) (1 .435ppm in Fig. 1 ) and 13 C-NMR signals at about 25.7ppm (25.773ppm in Fig. 2) indicative of the presence of a pyruvate group with a bridge bond between two hydroxyls.
  • the anomeric zone of the carbon spectrum may, moreover, preferably show from a minimum of 4 to a maximum of 6 signals, between 97 and 105ppm.
  • the polysaccharides obtained from fermentation of the C2B2 strain can moreover optionally also have other signals attributable to the probable presence of O-acetyl groups such as for example signals at 2.06ppm, 2.148ppm and 2.201 ppm in the 1 H spectrum (see Fig. 1 ) and signals at about 20ppm (20.958ppm in Fig. 2) and at about 174ppm (173.866ppm in Fig. 2) in the 13 C spectrum.
  • one or more sugars making up the polysaccharide can be bound to one or more pyruvate groups with a bridge bond between two hydroxyls, and moreover the polysaccharide can be in non-acetylated form or can have one or more acetyls on one or more monomers.
  • Fig. 1 and Fig. 2 show, respectively, the 1 H- and 13 C-NMR spectra of a polysaccharide (or mixture of polysaccharides) obtained as a result of aerobic fermentation of the C2B2 strain as described above.
  • the spectra were obtained using the model VXR300S NMR Spectrometer (Varian Inc., Palo Alto CA, USA).
  • Said polysaccharide (or mixture of polysaccharides), if submitted to acid hydrolysis by for example treatment with sulphuric acid at 50-96% (for example carried out at the reflux temperature for 30 min-10 hours) or treatment with trifluoroacetic acid (TFA) at concentrations of 10-60% at the reflux temperature for 20 min-2 hours (in the case of TFA it is possible to work at pressures above atmospheric, for example up to about 2 atm as is obtainable in autoclave)
  • a hydrolysis mixture is obtained consisting of the following principal monosaccharides, which can be separated by HPLC (Perkin Elmer with Transgenomic Column ICE SEP Ion 300, eluent 0.015 N sulphuric acid, column temperature: 40°C, flow: 0.4 ml/min). (see Fig. 4)
  • Fig. 5 shows the HPLC plot (Perkin Elmer HPLC with Transgenomic Column ICE SEP Ion 300, eluent 0.015 N sulphuric acid, column temperature: 40°C, flow: 0.4 ml/min) obtained as a result of partial acid hydrolysis.
  • ion exchange resins Rost and Haas, Amberlite IR 120 and Amberlite IRA 94S
  • the aforementioned hydrolysed polysaccharide therefore provides mixtures containing mainly L-fucose, D-glucose, D-galactose and preferably also glucuronic acid and pyruvic acid, and from these mixtures it is possible to isolate the desired L-fucose at high yields and purities.
  • the aforementioned polysaccharide After partial hydrolysis, the aforementioned polysaccharide has, according to HPLC analysis of the hydrolysate, preferably a molar content of glucose/galactose/fucose/pyruvic acid in the relative proportions 0.5 : 0.5 : 1 : 0.5
  • the aforementioned polysaccharide has, according to HPLC analysis of the hydrolysate, preferably a molar content of glucose/galactose/fucose/glucuronic acid/pyruvic acid in the relative proportions of 0.5 : 1 : 1 : 0.5 : 0.5.
  • the mixture at the end of fermentation is fluidized until a viscosity below 100 cP is reached, by adding an aqueous solution of a strong acid, preferably sulphuric acid, preferably with a concentration between 30 and 70 wt.%, up to a pH between 1.5 and 4.5, preferably between 2.0 and 3.0.
  • a strong acid preferably sulphuric acid
  • the acidified mixture is then heated at temperatures between 60 and 80°C, for times between 2 and 10 hours, preferably for 4-8 hours.
  • the first purification step is another crucial point of the process: the polysaccharide is recovered by ultrafiltration since it remains quantitatively in the retentate together with just the cells of the microorganism while all the fermentation by-products (e.g. butanediol, lactic acid etc.) and the salts and the acid added for fluidization are removed almost quantitatively in the permeate.
  • the fermentation by-products e.g. butanediol, lactic acid etc.
  • This step provides an excellent solution to the problem of purification of L-fucose, for obtaining a crystalline product of high purity; in fact it allows the sugars constituting the polysaccharide to be separated from all the low molecular weight by-products, for example butanediol, that cannot be removed in the steps described next, and that constitute a great problem for direct crystallization of the product, as they are present in considerable amounts.
  • the purification process is carried out on the retentate containing the polysaccharide according to known procedures (see for example EP102535, H. Voelskow and M. Schlingmann).
  • Complete hydrolysis is performed by treatment with a strong acid (for example sulphuric acid, hydrochloric acid, phosphoric acid, trifluoroacetic acid etc.). Sulphuric acid is preferred.
  • a solution of strong acid at 30-70 wt.% is added to the retentate and it is heated at the reflux temperature for 5-10 hours.
  • a base sodium hydroxide, calcium hydroxide etc.
  • Calcium hydroxide is preferred.
  • sulphuric acid and calcium hydroxide are used in combination: this leads to precipitation of CaS0 4 , which is removed by filtration or by centrifugation, thus removing a large part of the salts present in the reaction mixture.
  • D-glucose and D-galactose and glucuronic acid from the mixture is achieved by techniques known by a person skilled in the art, for example by chromatography or, preferably, by the action of a microorganism that utilizes said carbohydrates as a carbon source for growing, but leaves the L-fucose intact.
  • a microorganism is used and even more preferably the solution is inoculated with Saccharomyces cerevisiae.
  • Fermentation is conducted until glucose and galactose and glucuronic acid disappear completely.
  • the solution is then pasteurized at 60-70°C for 10-30 minutes and the yeast cells are removed from the solution of L-fucose by ultrafiltration or microfiltration, and the ultrafiltered or microfiltered solution is deionized by passing over strong cationic and weak anionic ion exchange resins in series.
  • the deionized solution is concentrated to a syrup and L-fucose (whose HPLC purity in the solution at this stage is above 75%) is crystallized by adding solvent, for example alcohols such as methanol, ethanol, n-propanol, isopropanol or 2-butanone.
  • solvent for example alcohols such as methanol, ethanol, n-propanol, isopropanol or 2-butanone.
  • the strain producing the polysaccharide containing L-fucose was isolated from 1 g of foliar homogenate of Phalaenopsis sp. (Orchidacea) in saline and diluted serially in the same solution to 10 ⁇ 8 . 1 ml of each dilution was seeded at depth in Petri dishes containing 15 ml of TSB medium (Trypticase Soy Broth, Biolife) solidified with 15g/l of agar and left to incubate at 30°C for three days.
  • TSB medium Trypticase Soy Broth, Biolife
  • the yellow pigmented colonies, clear and of large convex dimensions, that develop on the surface of the agar were then transferred to fresh TSB medium for purification by smearing them for three successive cycles, starting each time from a well isolated colony.
  • the strain was submitted to summary identification by the usual methods of biochemical characterization (API tests) and by sequencing the ribosomal RNA 16S (SEQ ID NC ).
  • the purified strain was stored in glycerol both at -20°C and at -80 °C.
  • HPLC analysis of the hydrolysate produces the following results (comparing only the content of glucose, galactose and fucose after hydrolysis of the fermentation broth without treatments of purification and isolation of the components)
  • Glucose:Galactose:Fucose The molar proportions of Glucose:Galactose:Fucose are thus found to be 1 .8 : 1 : 1 .3.
  • 1 10 I of culture medium is prepared, composed of:
  • Inoculate with 3 litres of preferment prepared according to Example 2 and leave to stand at 30°C, stirring vigorously, blowing-in air from below, at slight pressure, monitoring the glucose concentration.
  • glucose is absent, add 5.5 litres of a sterile solution of glucose at 50% w/v, monitoring the pH, which must be at 7.0 ⁇ 0.3, and the viscosity of the medium.
  • Glucose:Galactose:Fucose The molar proportions of Glucose:Galactose:Fucose are found to be: 3.4 : 1 : 2.2
  • the preferment is left at 30°C with stirring, blowing-in air from below and under slight pressure for about 24 hours.
  • the molar proportions of Glucose:Galactose:Fucose in the preferment are found to be: 3.1 : 1 : 1 .7
  • the fermentation takes a total of 140 hours.
  • the molar proportions of Glucose:Galactose:Fucose are found to be 2.7 : 1 : 2.0
  • Example 4 15 m 3 of ferment obtained from Example 4 (containing 28 kg of L-fucose) is ultrafiltered for the purpose of separating and concentrating the polysaccharide. 315 kg of 50% sulphuric acid is added to the retentate from ultrafiltration, maintaining the suspension under reflux for 8 hours in a suitable reactor, for complete hydrolysis of the polysaccharide.
  • the suspension is adjusted to pH 5.5-6.5, by adding 180 kg of calcium hydroxide.
  • the precipitate of calcium sulphate that forms after adding calcium hydroxide is removed by centrifugation.
  • the supernatant is then adjusted to 37 °C, and inoculated with 2 kg of lyophilized yeast Saccharomyces cerevisiae.
  • the solution is then pasteurized at 70°C for 30 minutes and ultrafiltered.
  • the ultrafiltration permeate is deionized on strong cationic and weak anionic ion exchange resins in series.
  • the fermentation of the C2B2 strain was carried out as in example 4.
  • the solution was then concentrated and diafiltered by ultrafiltration with a polymer membrane with a molecular cut-off of 10 000 Da.
  • Glacial acetic acid was added to the supernatant until a final concentration of 2% was reached. The sample was then heated at 100 ⁇ € for 1 hour.
  • the sample was centrifuged again at 13000 rpm for 30 minutes and, after dilution 1 :1 with water, it was diafiltered against demineralized water and concentrated by tangential ultrafiltration with a spiral-wound membrane with molecular cut-off of 20000 Da.
  • the concentrate was dried to constant weight in a rotary evaporator (Rotovapor, Buchi). 5 grams of sample was treated with trifluoroacetic acid (TFA) to a final concentration of 16% in autoclave (2 atm) at 121 °C for 20 minutes. The solution obtained was exchanged with 100 ml of demineralized water by means of the rotary evaporator (Rotovapor, Buchi) 5 times.
  • TFA trifluoroacetic acid
  • the sample was then submitted to HPLC chromatographic analysis using the Perkin Elmer series 200 HPLC chromatograph, chromatographic column Transgenomic ICE- SEP ION 300 equipped with similar precolumn and refractive index detector with thermostatically controlled cells.
  • chromatogram given in Fig.5 shows the HPLC plot obtained in the same conditions on the same sample after elimination of the peak at 11 by passing over ion exchange resins.
  • the fermentation of the C2B2 strain was carried out as in example 4.
  • the solution obtained was ultrafiltered by filtration on a ceramic filter with cut-off of 300 000 Da (0.05 micron) and the permeate was concentrated using a nanofiltration membrane (SR2 membrane).
  • SR2 membrane nanofiltration membrane
  • the polysaccharide was then precipitated with methanol (final concentration methanol :water of 3:1 ).
  • the precipitate was finally filtered on a paper filter, washed with a solution of methanol :water 3:1 and dried in a stove at 40°C for 8 hours.
  • the sample was then submitted to HPLC chromatographic analysis using the Perkin Elmer series 200 HPLC chromatograph, chromatographic column Transgenomic ICE- SEP ION 300 equipped with similar precolumn and refractive index detector with thermostatically controlled cells.
  • Peak Time ; Component Concentration Area Response i Amount Area
  • the fermentation of the C2B2 strain was carried out as in example 4.
  • the solution was then heated at 97 °C for 6 hours.
  • the sample was then submitted to HPLC chromatographic analysis using the Perkin Elmer series 200 HPLC chromatograph, chromatographic column Transgenomic ICE- SEP ION 300 equipped with similar precolumn and refractive index detector with thermostatically controlled cells.
  • chromatogram is shown in Fig. 7. (see printout below)

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Abstract

La présente invention concerne un procédé de production, à l'échelle industrielle, de L-fucose (ou 6-désoxygalactose) à partir de l'hydrolyse de polysaccharides pouvant être obtenus par une fermentation aérobie d'une nouvelle souche microbienne isolée appartenant à la famille des entérobactériacées.
EP11769803.5A 2010-09-13 2011-09-13 Procédé de production de l-fucose Withdrawn EP2616547A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
ITFI2010A000188A IT1405680B1 (it) 2010-09-13 2010-09-13 Processo per la produzione di l-fucosio.
PCT/EP2011/065825 WO2012034996A1 (fr) 2010-09-13 2011-09-13 Procédé de production de l-fucose

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EP2616547A1 true EP2616547A1 (fr) 2013-07-24

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IT (1) IT1405680B1 (fr)
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Families Citing this family (35)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ITFI20120052A1 (it) * 2012-03-13 2013-09-14 Inalco Spa Processo per il recovery di l-fucosio da esopolisaccaridi.
EP2676536A1 (fr) 2012-06-22 2013-12-25 AIT Austrian Institute of Technology GmbH Procédé de production de graine de plante contenant des micro-organismes endophytes
DK2728009T3 (da) 2012-10-31 2017-11-06 Jennewein Biotechnologie Gmbh Fremgangsmåde til fremstilling af monosaccharider
EP2954043B1 (fr) 2013-02-05 2021-07-28 University of Saskatchewan Symbiotes microbiens endophytes en soin prénatal des plantes
WO2014210372A1 (fr) 2013-06-26 2014-12-31 Symbiota, Inc. Populations d'endophytes provenant de semences, compositions et procédés d'utilisation
US10136646B2 (en) 2013-06-26 2018-11-27 Indigo Ag, Inc. Agricultural endophyte-plant compositions, and methods of use
EP3659414A1 (fr) 2013-09-04 2020-06-03 Indigo Ag, Inc. Compositions agricoles associant plantes et endophytes et leurs procédés de préparation
CN105683387B (zh) 2013-09-06 2020-02-21 格礼卡姆股份公司 寡糖的发酵生产
HUE048814T2 (hu) 2013-11-06 2020-08-28 Texas A & M Univ Sys Gombaendofiták növelt terményhozamok és károsítók elleni védekezés céljából
EP3086646A4 (fr) 2013-12-24 2017-08-09 Indigo AG, Inc. Plantes contenant des endophytes bénéfiques
US9364005B2 (en) 2014-06-26 2016-06-14 Ait Austrian Institute Of Technology Gmbh Plant-endophyte combinations and uses therefor
WO2015100432A2 (fr) * 2013-12-24 2015-07-02 Symbiota, Inc. Procédé de propagation de micro-organismes dans des bioréacteurs végétaux et de stockage stable de micro-organismes dans des graines agricoles
MX367032B (es) 2014-06-20 2019-08-02 The Flinders Univ Of South Australia Inoculantes y metodos para su uso.
WO2015200902A2 (fr) 2014-06-26 2015-12-30 Symbiota, LLC Endophytes, compositions associées et leurs méthodes d'utilisation
DE202015009775U1 (de) * 2014-12-16 2020-02-05 Glycom A/S Abscheidung von 2'-FL aus einer Fermentationsbrühe
CN108271339A (zh) 2014-12-30 2018-07-10 靛蓝农业公司 栽培品种和物种间的种子内生菌、相关组合物及其使用方法
EP3050973A1 (fr) 2015-01-30 2016-08-03 Jennewein Biotechnologie GmbH Procédé de fermentation pour la production de monosaccharides sous forme libre à partir de sucres activés par des nucléotides
DK3274465T3 (da) 2015-03-26 2022-10-24 Basf Se Biokatalytisk fremstilling af l-fucose
BR112017023549A2 (pt) 2015-05-01 2018-07-24 Indigo Agriculture Inc composições de endófito complexo isolado e métodos para melhorar características de planta.
RU2017141758A (ru) 2015-05-01 2019-06-03 Индиго Агрикултуре, Инк. Разработанные комплексные эндофитные композиции и способы улучшения признаков растений
WO2016200987A1 (fr) 2015-06-08 2016-12-15 Indigo Agriculture, Inc. Compositions d'endophyte de genre streptomyces et procédés pour l4amélioration de caractéristiques agronomiques dans les plantes
US11751515B2 (en) 2015-12-21 2023-09-12 Indigo Ag, Inc. Endophyte compositions and methods for improvement of plant traits in plants of agronomic importance
US11312741B2 (en) 2016-04-19 2022-04-26 Glycom A/S Separation of oligosaccharides from fermentation broth
AU2017366699A1 (en) 2016-12-01 2019-07-18 Indigo Ag, Inc. Modulated nutritional quality traits in seeds
EP3558006A1 (fr) 2016-12-23 2019-10-30 The Texas A&M University System Endophytes fongiques pour l'amélioration des rendements des cultures et une protection contre les nuisibles
MX2019010350A (es) 2017-03-01 2019-12-19 Indigo Ag Inc Composiciones de endofitos y metodos para mejorar las caracteristicas de las plantas.
MX2019010349A (es) 2017-03-01 2019-11-21 Indigo Ag Inc Composiciones de endofitos y metodos para mejorar las caracteristicas de las plantas.
US11882838B2 (en) 2017-04-27 2024-01-30 The Flinders University Of South Australia Bacterial inoculants
US11263707B2 (en) 2017-08-08 2022-03-01 Indigo Ag, Inc. Machine learning in agricultural planting, growing, and harvesting contexts
WO2019057958A1 (fr) 2017-09-22 2019-03-28 Technische Universität Graz Particules polymères contenant des micro-organismes
EP3486326A1 (fr) * 2017-11-21 2019-05-22 Jennewein Biotechnologie GmbH Procédé de purification d'acide n-acétylneuraminique à partir d'un bouillon de fermentation
WO2020154217A1 (fr) * 2019-01-21 2020-07-30 Dupont Nutrition Biosciences Aps Procédé de fabrication de l-fucose
WO2021196572A1 (fr) * 2020-03-30 2021-10-07 西南大学 Polysaccharide extracellulaire riche en fucose, son procédé de préparation et son application
CN114835766A (zh) * 2022-05-20 2022-08-02 中国科学院力学研究所 一种从二元混合物中分离l-岩藻糖的方法
CN117646048A (zh) * 2023-12-08 2024-03-05 重庆智合生物医药有限公司 一种缩短岩藻多糖发酵时间的生产方法

Family Cites Families (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4298691A (en) 1980-09-05 1981-11-03 Merck & Co., Inc. Preparation of heteropolysaccharide S-156
DE3300633A1 (de) * 1982-08-10 1984-03-01 Hoechst Ag, 6230 Frankfurt Verfahren zur herstellung von rhamnose oder fucose
US4806636A (en) * 1985-03-20 1989-02-21 The Dow Chemical Company Heteropolysaccharide produced by Enterobacter sakazakii
FR2729969B1 (fr) * 1995-01-26 1997-04-18 Bieurope Nouvelle souche de klebsiella pneumoniae, subsp. pneumoniae et procede de production d'un polysaccharide contenant du l-fucose
US6713287B1 (en) 1995-10-24 2004-03-30 The Scripps Research Institute Enzymatic synthesis of L-fucose and L-fucose analogs
KR100397793B1 (ko) * 2000-07-28 2003-09-13 삼성전자주식회사 느릅나무로부터 분리된 신규의 미생물 및 그를 이용한항암 면역활성의 세포밖 당화합물의 제조방법
FR2840920B1 (fr) * 2002-06-18 2006-06-23 Biochimie Appliquee Soc Nouveau microorganisme de la famille des enterobacteriaceae
FR2842201B1 (fr) * 2002-06-18 2005-01-14 Probest Nouvel oligosaccharide, compositions cosmetiques et/ou dermatologiques en contenant et ses applications
JP2004208563A (ja) * 2002-12-27 2004-07-29 Asahi Kasei Corp 新規微生物、水溶性多糖類及びその製造法
PT104888B (pt) * 2009-12-15 2020-09-10 73100 - Setenta E Três Mil E Cem, Lda Biopolímero bacteriano contendo fucose

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
FILOMENA FREITAS ET AL: "Fucose-containing exopolysaccharide produced by the newly isolated Enterobacter strain A47 DSM 23139", CARBOHYDRATE POLYMERS, APPLIED SCIENCE PUBLISHERS, LTD. BARKING, GB, vol. 83, no. 1, 23 July 2010 (2010-07-23), pages 159 - 165, XP002638650, ISSN: 0144-8617, [retrieved on 20100723], DOI: 10.1016/J.CARBPOL.2010.07.034 *
See also references of WO2012034996A1 *

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