Classifications

• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
  • C12G1/00—Preparation of wine or sparkling wine
  • C12G1/06—Preparation of sparkling wine; Impregnation of wine with carbon dioxide
• • 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
  • C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
  • C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
  • C12N11/10—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a carbohydrate

Definitions

• the invention relates to a process for obtaining biocatalysts in which a core containing microorganisms is coated with a coating of gelable and / or crosslinkable compound.
• biocatalysts of this type can be used for the preparation of fermented drinks such as wine and beer. Their use has also been proposed for the manufacture of yogurts and cheeses.
• the object of the invention is therefore to provide a process for the preparation of biocatalysts with a high content of active dry matter, which can be used on an industrial scale.
• biocatalysts in dry form which can be stored for long periods at room temperature and which can be used without prior rehydration.
• the process for manufacturing biocatalysts of the invention is characterized in that it comprises the coating of particles whose dry matter content is greater than approximately 30%, containing active microorganisms, and their drying so as to obtain a rate residual moisture not exceeding about 10%, preferably 4%.
• particle denotes granules, pellets, spherules, beads, that is to say any agglomerate of microorganisms.
• the high dry matter content of the particles used in the coating step which is advantageously greater than 50%, promotes the subsequent drying of the coated particles and makes it possible to easily reach contents of the order of 90 to 96 % dry matter, while maintaining a survival rate of almost 100% of microorganisms.
• This survival rate is controllable after release of the microorganisms by counting on a suitable medium in Petri dish or, for yeasts, by microscopic observation of the discoloration of methylene blue.
• This process is more particularly characterized in that the said particles are produced from preparations of microorganisms whose dry matter content is such that these preparations are extrudable.
• preparations obtained by concentration from the culture of these microorganisms for example by centrifugation and / or filtration and / or pressing.
• the particles used in the coating step advantageously contain, in addition to the microorganisms, compounds capable of increasing the dry matter content of the core intended to be coated.
• these compounds are chosen more especially from products acceptable in the food industry and in particular in fermentation processes.
• binders or lubricants are chosen so as to promote the integrity of the microorganisms during extrusion.
• products acceptable in the food industry there will be mentioned carrageenans, gelatin and ovalbumin.
• the use in the process of the invention of particles comprising the additives mentioned above makes it possible to bring their level of dry matter to at least 35 to 50% approximately and to facilitate the extrusion.
• the active microorganisms are mixed with low humidity, that is to say that unlike many embodiments of the prior art, they are not immobilized in a gel or other medium with high water content.
• particles are used as obtained by extrusion.
• the extrusion step is carried out so as to obtain particles having a size giving a satisfactory exchange surface for the applications envisaged.
• Appropriate pellet dimensions correspond to diameters of 0.5 to 3 mm.
• the particles of microorganisms with, where appropriate, the additives mentioned above are obtained by the rotor granulation technique.
• a spheronization step during which it is also possible to subject the particles to a pre-drying phase.
• a spheronizer comprising a rotating embossed tray, drying is carried out after the spheronization step.
• the establishment of a fluidized bed in the spheronizer allows drying of the particles at the same time.
• temperatures of 40-50 ° C are more specifically applied in order to impart a temperature of approximately 30 ° C to the particles at the outlet, which makes it possible to avoid any deterioration of the cells.
• a gelable and / or crosslinkable product of low viscosity that is to say sprayable by nozzles so as to create a regular layer around the particle containing the microorganisms and where appropriate the additives. mentioned upper.
• Appropriate viscosities are less than approximately 200 ⁇ 10 ⁇ 3 Pa.s and in particular of the order of 5 to 10 10 ⁇ 3 Pa.s.
• the coating of the particles is preferably carried out by spraying said product, according to the technique called spray-coating, onto the particles which are advantageously pre-dried.
• the particles are suspended in a stream of hot air while the coating solution is vaporized. Passing through the coating cycle several times makes it possible to uniformly cover the surface of the particles.
• the temperature and the flow rate of the air, as well as the flow rate and the spray pressure, are chosen so as to form a homogeneous layer and to avoid the formation of flakes.
• the sprayed products are subjected to hardening or crosslinking in order to ensure the mechanical integrity of the biocatalysts formed and to prevent the release of microorganisms.
• the usual techniques are followed, most generally by physical or chemical means.
• the biocatalyst spherules are immersed in a bath containing the crosslinking agent.
• coated particles are subjected to a drying step. This step is carried out in a fluidized bed.
• the particles are subjected to a preliminary drying until a final humidity of 6 is obtained. at around 4%. These particles are then coated in a stream of cold air in a fluidized bed.
• the process of the invention is particularly suitable for coating microorganisms such as yeasts, bacteria, molds.
• yeasts there will be mentioned the genus Saccharomyces and in particular Saccharomyces cerevisae.
• the bacteria which can be used for the preparation of the biocatalysts defined above include the genera Lactobacillus and Streptococcus and in particular Lactobacillus casei and Streptococcus lactis.
• Usable molds include the genus Af-pp.rgillus and, in particular, Aspergillus niger.
• biocatalysts in which the microorganisms retain excellent viability over time, at room temperature, as evidenced by the controls carried out according to the techniques mentioned above.
• These biocatalysts can be used directly without a rehydration step.
• new products they are also part of the invention. They are more particularly characterized in that they comprise a core of active microorganisms, optionally added with fillers and / or binders as indicated above, coated with an external layer of gelable and / or crosslinkable compound, and that their dry matter content is more than around 90%.
• biocatalysts are in the form of beads or pellets of approximately 0.5 to 3 mm in diameter.
• Example 1 Manufacture of biocatalyst containing S.cerevisae coated with a crosslinking agent.
• the manufacturing process used includes the following steps:
• the yeast preparation obtained is mixed with the filler products and the binder at a very high concentration, so as to obtain a final mixture of 40 to 50% of dry matter (basic formula).
• the mixing is carried out in. a mixer for 20 to 30 min.
• the extrusion is then carried out using an extruder (ALEXANDER WERK S12 M70R) by passage through a cylinder covered with orifices 1 mm in diameter, the extrusion speed is fixed at 375 rpm.
• the pellets obtained are spheronized on an embossed tray, for example by passing through a KALEVAR model 15 spheronizer for 2 to 20 min at 800 rpm. 3) Pre-drying
• the granules obtained are pre-dried on a Niro aeromatic MPlR fluidized air bed equipped with a spraying device. 1 kg of granular product is placed on a perforated plate, a current of hot air passes through this support from bottom to top with a flow such that the product is fluidized.
• the duration of this pre-drying is 10 to 40 min. This is uniform thanks to the large air / product contact surface and intense agitation.
• the most suitable inlet air temperatures are between 40 and 50 ⁇ C. It is important that the yeasts are kept as low as possible, namely 25 to 33 ° C.
• a residual water content of 35 to 8% can be reached in the microorganism granules after a residence time in the fluidized bed of 40 min.
• the survival rate after drying is close to 100%.
• An outer layer is produced by coating in a fluidized bed the pre-dried granules of microorganisms, which makes it possible to increase the mechanical resistance of the granules and to prevent the release of microorganisms during the use of the granules.
• the coating in a fluidized bed is carried out by spraying (spray coating) on the granules suspended in an upward current of hot air. A coating solution is then sprayed through a nozzle.
• the polymer solution used is, for example, a low viscosity alginate of 5 to 15% dry matter.
• the particles pass several times per minute in the coating cycle, which allows according to the invention a uniform distribution of the coating solution on their surface.
• the coating conditions are as follows:
• the particles coated in a fluidized bed are then soaked in a CaCl 2 solution at 30 g / l.
• Crosslinking is initiated immediately and after 60 minutes of contact, the particles including the cells of microorganisms are rinsed with running water.
• the washed beads are drained, then dried in a fluidized bed type NIRO aeromatic MP1 R up to 40 to 50% of dry matter. Drying is carried out at a programmable air inlet temperature of 30 to 50 ° C and at an air flow rate which corresponds to a pressure drop of 150 mm of water.
• the final humidity does not exceed 3 to 8% after 45 min of drying.
• These beads can be used as such without rehydration and without the need to re-acclimatize the microorganisms.
• biocatalysts make them particularly suitable for the preparation of fermented drinks or the manufacture of yogurts and cheeses.
• the foam is taken in the bottle.
• the yeasts which ensure the second alcoholic fermentation are added to the sweet base wine when the wine is drawn at a concentration of 1-1.5 million cells per milliliter.
• yeasts form a sedimentation deposit.
• the riddling operation which consists in gathering this deposit in the neck of the bottle requires a significant investment in material, place, time and handling.
• biocatalysts of the invention makes it possible to simplify these operations. These are used, stored at room temperature, in a sachet, under vacuum or in a controlled atmosphere.
• Example 3 An example of application is given below.
• Example 3 An example of application is given below.
• biocatalysts prepared according to Example 1 is introduced, without prior rehydration, into a bottle containing a filtered wine sweetened to approximately 24 grams per liter.
• the use of the beads without rehydration allows their distribution in the bottle by a volumetric type doser adaptable on most bottling groups.
• the bottle After the draw, the bottle is placed on slats.
• the foaming takes place under temperature conditions and with fermentation kinetics equivalent to a free yeast draft.
• the yeast balls denser than wine, are collected in the neck, in less than a minute, by simply passing the bottle from the horizontal position to the vertical position, neck down, called “on point ".
• the clarity of the wines is preserved, taking into account the characteristics of the beads of the invention.

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• Organic Chemistry (AREA)
• Life Sciences & Earth Sciences (AREA)
• Chemical & Material Sciences (AREA)
• Health & Medical Sciences (AREA)
• Engineering & Computer Science (AREA)
• Bioinformatics & Cheminformatics (AREA)
• Zoology (AREA)
• Wood Science & Technology (AREA)
• Genetics & Genomics (AREA)
• Biochemistry (AREA)
• General Health & Medical Sciences (AREA)
• General Engineering & Computer Science (AREA)
• Molecular Biology (AREA)
• Biomedical Technology (AREA)
• Biotechnology (AREA)
• Microbiology (AREA)
• Immobilizing And Processing Of Enzymes And Microorganisms (AREA)

Applications Claiming Priority (3)

Publications (1)

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Family Applications (1)

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Family Cites Families (4)

• 1993
  • 1993-07-06 FR FR9308285A patent/FR2708281B1/fr not_active Expired - Fee Related
• 1994
  • 1994-07-06 AU AU72593/94A patent/AU7259394A/en not_active Abandoned
  • 1994-07-06 HU HU9503844A patent/HU9503844D0/hu unknown
  • 1994-07-06 WO PCT/FR1994/000834 patent/WO1995002047A1/fr not_active Application Discontinuation
  • 1994-07-06 EP EP94921692A patent/EP0707643A1/de not_active Withdrawn
  • 1994-07-06 BR BR9406920A patent/BR9406920A/pt not_active Application Discontinuation
  • 1994-07-06 CA CA002166602A patent/CA2166602A1/fr not_active Abandoned
  • 1994-07-06 ZA ZA944894A patent/ZA944894B/xx unknown

Non-Patent Citations (1)

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