LU86023A1 - SYSTEM AND APPARATUS FOR FERMENTATION IN FLEXIBLE TANKS - Google Patents

Description

i - i - j i! *

The present invention relates to a system for fermenting initially sterile liquids, by microorganisms in pure culture, under conditions preventing any infection by other microorganisms, whether the fermentation is continuous or not, a method using one or more - * their flexible tanks arranged in series or in parallel, initially sterile or completely sterilized, as well as the apparatus allowing it to operate.

It also relates to the refermentation of liquids not recontaminated by undesirable microorganisms, as well as the culture of microorganisms.

The flexible reservoir (s) are made up of flexible, flexible envelopes of generally arbitrary shape, but above all cylindrical and terminated by hemispherical or other sections, immersed in a liquid which may be water in which there is , or not, dissolves or disperses solids, liquids or salts, in order to give it desirable properties.

In its simplest execution, the system and the equipment enabling it to operate comprises only one flexible tank per fermenter.

25 In Figure 1 we have 1 s Flexible tank.

2 s Liquid in which the flexible tank is immersed.

3 s Gaseous atmosphere, at atmospheric pressure, * 30 between the liquid 2 and the walls of the enclosure covering the basin 4.

"4: Basin containing the liquid 2.

5: Rigid or flexible roof (optional if the atmosphere 3 is air) j 35 6: Aseptic circulation pump.

; 7: Arrival of the sterile liquid to be fermented.

8; Aseptic arrival of sterile air or oxygen. and a rigorously pure culture of the microorganism (s) used. ft I * 'ff - 2 - 9: Aseptic output of the fermented liquid.

10: Exit of the gas produced by fermentation.

11: Device avoiding any contamination of the contents of the flexible tank 1 by microorganisms-5 other than those which were initially introduced in 8.

12: Devices maintaining the desired characteristics in the liquid 2. (temperature, oxygen content *...) 10 13: Net, false bottom, or distribution network of the immersion liquid used to carry out local deformations of the membrane flexible tank.

The entire system is sterilized before commissioning and, except in the event of an accident, should not be sterilized during its lifetime, provided that the following products: liquid to be fermented, as well as air or oxygen are thoroughly sterilized by known methods before entering the system, regardless of whether the microorganism culture is; rigorously pure and aseptically introduced (into the system, that the withdrawals of fermented and stored liquids are carried out aseptically! , as well as the evacuation of the fermentation gases do not allow any microorganism different from that used to be introduced into the liquid undergoing fermentation 30 controlled by a selected microorganism.

,; The flexible reservoir, initially empty and sterile, is aseptically supplied with liquid ·! sterile and in pure culture and oxygen gas or sterile air if necessary given the type of microorganism and therefore fermentation used, the! sterile liquid with low gas content.

i The liquid is agitated, for example by pumping in a closed circuit. It is maintained at the desired temperature 40 and remains completely protected from parasitic fermentations while the fermentative gases

Jb - 3 - if necessary, these are evacuated from the upper part of the flexible tank to the outside or to a recovery installation via a device eliminating any risk of reinfection of the contents of the flexible tank.

The microorganisms remain dispersed in the flexible tank, the contents of which are in movement, while the deposits of significant thickness on the flexible membranes of the tank are regularly detached thanks to the external action on these flexible membranes of localized forces causing their deformation; these localized forces being carried out mechanically or even hydraulically by means of localized jets implements the liquid in which the flexible tank is immersed.

The circulation pump ensuring the movement of the liquid during fermentation is of the high flow, low pressure type; 20 or else of the low flow, high pressure type, if the content of the flexible tank is kept in motion indirectly via a! hydroejector, but this pump is preferably an aseptic pump of a new type described below (fig.II) or similar to the latter.

In figure II we have: 1 s Motor 2 s Pump shaft i * I 3: Pump jam | 30 4: Pump body! 5: Arrival of the pumped fluid! · 6: Start "'7: Antiseptic tank 8: Antiseptic 5 35 9: Mobile platform 7' and 9 'î Positions of 7 and 9 when the pump is no longer immersed in the antiseptic, 1. during a repair and sterilization; 'tion by heat.

i

II

- 4 - I *

The elements 3 - 4 - 5-6 of the pump are sterilized by all conventional means, including 5 with water or steam containing, or not, a product ensuring the destruction of microorganisms, after which the tank 7 containing a ex-, this stable and cold antiseptic 8 is raised thanks to the platform 9, so that 10 the sterile elements of the pump by which a reinfection would be possible in a few days, months or years are immersed in a quantity important cold antiseptic.

This is chosen from a large number of possible compositions so that it cannot, if traces of it penetrate into the liquid conveyed by the pump, harm the organoleptic qualities of this liquid or the health of consumers.

20 The aseptic pump described in FIG. II

unsuitable when the hourly flow is very! *:? - 5 - high, of the order of 500 to 1000 m and more.

For very large flow rates the diameter of the pipes is 0.5 to 1 meter, or even greater and, moreover, it is then preferable to have the circulator built on plans.

In such cases the mobile container (7) is not suitable. It is replaced by a large chamber or tank.

The pump or the pipe comprising a propeller, its shaft and the packing, as well as the coupling flanges with the rigid pipe network, are sterilized.

Thereafter, antiseptic is immediately poured onto the parts, normally very hot, sterile, through which reinfection would be possible in the event of a micro-leak, then the tank is quickly filled with anti-septic, thus becoming the equivalent of tray 7 of Figure II.

20 Very large waterproof valves are not necessary when, as a repair is required, it is important to isolate the giant pump from the flexible tanks, since the pressure in the liquid network does not exceed 0.5 K ° / cm, it suffices to crush two flexible sections of the liquid transfer network i, one downstream and the other upstream of the circulator, in order to be able to isolate the latter. It then remains to appropriately re-sterilize the fraction of the installation which has been infected and to restart the circulator by ensuring total, "and permanent aseptic thanks to the antiseptic.

! - If necessary, the pressure outside the flexible sheaths can be maintained hydraulically so that there is pressure balance between the inside and the outside of the sheath.

; The simultaneous use of flexible sheaths and sterilization by fluent steam requires | that the pump has a steam inlet, 40 a condensate outlet and an ali device. mentation in sterile inert air or oaz. lu I 11

P

•;? - 6 -

The simplest antiseptics that may be suitable are concentrated solutions of sodium or potassium hydroxides as well as strong acids such as sulfuric acid, but dozens of other compositions can also achieve aseptic action. 'appa-; inexpensive reils, not having been specially constructed with a view to operating aseptically, and having one or more possibilities of micro-leaks allowing the reinfection of the product circulating in the apparatus. The latter can be a pump, a disperser, a colloid mill, a continuous centrifugal separator and even a simple connection by flanges or removable rac-cords.

A remarkable characteristic of this new technique is to ensure the maintenance of aseptic for extremely long periods, in a very simple, reliable and inexpensive manner, the elimination of the risks of reinfection being total insofar as one monitors the volume and composition of the bactericidal immersion fluid from time to time.

This new technique, necessary for devices which are not available in aseptic execution, presents a major advantage with respect to so-called aseptic embodiments because the aseptic is completely guaranteed and requires only a minimum of maintenance while the organs responsible for aseptic 30 so-called aseptic achievements require maintenance followed by sterilization whose frequency varies between once a day and once every 50 to 100 days, frequencies involving risks and periods of downtime not admissible when it is important to carry out the fermentations in question.

The use of flexible tanks makes it possible to avoid any movement of gas during successful filling and emptying operations. Anyone skilled in the art knows how much he is

difficult and insecure to sterilize M flows

- 7 - significant gas. This problem is eliminated.

The immersion of the flexible tanks in a liquid leads to the following advantages: s Reduction of the stresses to which the envelope is subjected and, in view of this, possibility of producing immense flexible tanks for 4 liquids.

- Protection against voluntary or accidental depredations, human or animal.

10 - Easy temperature control.

- Together with an adequate shape of the surface on which the tank is placed, ensures an almost constant pressure, facilitating the emptying of stored liquids.

15 - Reduction or elimination of the oxidation of the contents of the tanks.

- Easy suspension of deposits liable to adhere to the flexible walls carried out hydraulically by means of localized jets of immersion liquid or by mechanical action.

Regarding oxidation, the membrane forming the wall of flexible tanks will be chosen according to the requirements of the liquid and the fermentation it undergoes.

The permeation of oxygen through the membrane of the flexible tank and of the flexible or rigid sheaths or pipes which are connected to it and where the liquid circulates is the only source of involuntary oxidation of the liquid in fermentation.

Depending on the type of fermentation the liquid needs extremely variable amounts of oxygen, ranging from zero to a lot. The "zero" corresponds to strict anaerobic microorganisms and the "many" to strict aerobic microorganisms, then.

35 that, for alcoholic fermentation, economical! very important, the quantities of oxygen re-

quises are weak. In the latter case, it is generally advisable for there to be no supply of sterile air or oxygen in well-defined quantities 40 which during the periods during. ‘The fermented liquid is not withdrawn from the tank at A

1 t '* r - 8 - put on sale. This rule is not, however, absolute because, even for finished beer, the air of which is a formidable enemy, some very dark beers, of high fermentation, such as Stout, 5 can be improved by the presence of traces of oxygen. .

When it is imperative to reduce or eliminate oxidation, the process which is the subject of the present invention implements me or more of the following 10 techniques: - Total or partial reduction, by physical, chemical or biological processes of the oxygen content of the immersion liquid.

Protection or covering of the surface of the liquid eliminating or reducing the rate of dissolution of the gases in the immersion liquid.

- Replacement of the air on the surface of this liquid by an oxygen-depleted or oxygen-free gas.

20 - The influence of the immersion liquid on the tension in the membrane is very important.

Either a 300 m3 tank, 5 m diameter. height when filled 3 m. (80% degree of filling), density of the liquid to be fermented: 1.08.

25 - The tank being in the open air, we would have the same voltage: DH2 _ 1.08 x 3002 „1 _ ~ 4 --- 4- X IÖÖÖ ~ 24.3 Kg / Cm · - The tank being immersed in l water we have: DH2 _ 0.08 X 3002 „1 _,„ / 2 "4 --- 4- X 1000" 1.8 Kg / Cm 30 .- The tank being immersed in water whose weight specific was brought to 1.07 the tension would be: DH2 _ 0.01 X 300 2 .. 1 _ „2 -4 --- 4-x ΐοδο" 0.225 Kg / cm

Ultimately, if the densities are identi-35 c inside and outside the flexible tank, the latter retains its initial shape, cylindrical d- B the example considered and the flexibility of its walls is. maximum given the absence of J

b - 9 - * voltage. In addition, under these conditions, the tank can be filled to 100%.

The process certainly makes it possible to carry out discontinuous fermentations, for which the tank is filled, allowed to ferment and then transfers the fermented liquid to other tanks, similar or different. This procedure is necessary for certain fermentations.

In a brewery it is generally preferable to carry out a continuous fermentation or close to it and then, for an initial density, already high of, for example, 1.072, the fermented beer in circulation at a density of the order of 1.015.

2 15 - Tension in the membrane: 0.3375 K / cm.

- Cacl2 to add to the pond water to balance approximately the densities + 2%.

- The choice of membrane and degassing treatment for the immersion liquid depends on the degree of anaerobiosis to be maintained in the fermenting liquid.

With a 2 mm high density polyethylene membrane. thick and water in motion, saturated with oxygen, the permeability of the membrane 25 is P = 3.3 v ° * 3 = 3 * 2 cm3 of oxygen / day / m3. u 2

If a 300 m tank receives 60 m3 of dense wort saturated with oxygen per 24 hours, this represents about 540 grams or 378,000 cm. oxygen, while the membrane can only let 30 pass around 1181 cm. Therefore, gaseous oxygen must be injected into the beer stream for the reproduction of the yeast to be sufficient.

On the other hand, if the tank is used to carry out a very slow secondary fermentation there, the choice of a membrane which is not very permeable is indicated.

, With a membrane of reduced permeability,

type 2 mm. in high density polyethylene with barrier layer in Clarene L. (Solvay) or in polyvinylidene chloride 40, a minimum permeability is reached I

of the membrane. ‘I

λ r ’· - 10 -

Permeability of barrier product, -4 3

Clarene L, being 6 x 10 cm / cm / day / atmosphere / m2, that of a 0.01 mm clarene film.

✓ -4 3 3 thick is 6x10 xlO x 0.2 = 0.12 cm / 2 5 day / m and that of the whole is calculated as follows: 1 _ 1 d. 1 _ 0 P "372 + 0 ^ 2” 8'646 Hence P = 0, li5 cm8 / day / m ^ 3 and therefore 42.45 cm of oxygen per day and per 300 m tank, figure To improve the anaerobiosis further, the thickness of the barrier product can be increased or the oxygen content of the immersion liquid reduced.

In practice, it is better to reduce the oxygen supply by the members as much as possible. ._____ 15 born because, then, the same installation can also be used for fermentations requiring anaerobiosis or for the aseptic storage of oxygen-sensitive products such as fruit juices.

In fact flexible fermentation tanks 20 are easily transformed into aseptic storage tanks for sterile liquids, the reverse being more difficult.

When it is important to make the quantities of oxygen passing through the flexible membranes absolutely negligible, the immersion liquid is degassed and covered with a barrier film or a gas very low in oxygen, or accumulates these precautions.

Or: High density polyethylene membrane, 2 mm.

30 Clarene barrier layer L 0.01 mm.

Degassed water in balance with one; gas containing 1% oxygen.

Permeability of the polyethylene layer: 3.2 x 0.01 _ Λ 3, v, 2 -0 "2--0.16 cm / oxygenated / day / m.

35 Permeability of the Clarene layer: —4 3 v 2 6 x 10 x 10 x 0.01 = 0.006 oxygen / day / m.

Water permeability. ’Not considered because it is in motion.

Permeability of the assembly: We have ii + 1.40 - -ar ". ^" ........ = 173. So P = O .OOR 7 R · pm2 / nwnpnp /. -11 _ 3 days / m2, and for a 300 m tank, approx. 2.13 cm ^ of oxygen per day Oxidation is zero.

It is easy to remove the oxygen from the cycled water by cooling it by vacuum or by displacing this gas with a neutral gas such as carbon dioxide.

It is preferable to supply rigid tanks with deaerated and / or chilled water from below. These problems arise from the usual techniques, as well as those relating to the insulation of the tanks, obtaining an atmosphere very poor in oxygen and the construction of a mattress or film floating on the surface of the immersion liquid and limiting the contact between this liquid and the atmosphere.

Buffer tanks, flexible, having only to carry out the aseptic storage of 20 sterile fermentable products in many cases can be sterilized before their placement. The same is true for certain flexible tanks, pipes and apparatuses used in the context of the present invention, while other installations are preferably designed with a view to being sterilized before their installation and able to be re-sterilized each time When it is useful, for example, to use a new pure culture of the microorganism under consideration, or another type of microorganism.

It is always advantageous to be able to sterilize at fairly high temperatures, above 100 ° C., the interior of the pipes and devices resistant to this treatment, whereas for flexible tanks and plastic sheaths which can deform freely, it is necessary to do not exceed atmospheric pressure, the temperature being 99 - 100 ° C maxi

i - 12 - mum, we can devote to these rare operations the time it takes / however there is interest, rather than maintaining for example 100 ° C for 20 h. with a view to destroying the most heat-resistant spores, increasing the lethality of steam or moist heat using one or more conventional chemicals such as strong acids,.-alkalis, formaldehyde, hydrogen peroxide, halogens and chlorine or iodine derivatives, quaternary ammonium, nitrites, sulfurous acid.

The germicide is introduced in gaseous or mist form in the vapor flow or else of a mixture "sterile hot gas, brouil-15 hot water" if it is necessary to work at less than 100 ° C.

The bactericidal products are then completely eliminated. The easiest way is to use pure steam for membranes, sheaths and 20 plastic or elastomer accessories resistant to 100 ° C. In this case, the final elimination of a powerful antiseptic, such as formaldehyde, is obtained by sweeping carried out using pure steam. This removes the antiseptic gas phase while the pure vapor condensate rinses the surfaces and flushes out impure liquid residue.

The effectiveness of the treatment requires that a slight regular slope exists between the basin of the first flexible tank and the final purge orifice 30 provided with a sealed aseptic valve.

A new technique for sterilizing mixed conduits, flexible sheath - pipes or device, rigid - flexible sheath, consists in sterilizing in an autoclave, under pressure, the rigid element.

35 Figure m describes the required sterilizer

We have 1 and 4: Flexible sheath.

2 i Rigid PIPE or rigid device.

3: Valve and possible air filter.

40 5: Autocia 6 ï Autoclave cover. j | - 13 - 7: Arrival of a sterile gas.

8: Pliers sealing the flexible sheath.

If the sheath is sterile at the start, the pins 8 are placed and then the rigid element 2 is mounted between 5 the re-infected parts.

The purge valve 3, terminated by a gas-tight filter 9 'which is permeable to microorganisms being open, the autoclave is closed and the sheath 1 and 2 are crushed, between this and the au-10 toclave cover, then sterilizes under pure steam pressure.

It is slowly depressed and lets in air or neutral gas, as the case may be, at 7.

A similar technique makes it possible to sterilize or maintain sterile sets of devices such as valves and the pipes connecting them, but then the autoclave is of the fig.iy type or one has: 1-2-3 s Sealed valves , automatic or manual.

4: Autoclave body.

20 5: Autoclave cover.

6 î Jams, flanges, welds, etc. .

7 i Jam allowing 5 to move to 4.

This device is especially advantageous for the 25 sterilization and aseptic cooling groups, whatever the fluid treated, because it eliminates the risks of reinfection and also keeps the interior of the watertight valves, with membrane by a high temperature. example.

30 They are a potential source of reinfection as soon as the downstream part of the membrane ceases to be sterile.

When the product is sterilized at more than 100 ° C and the devices at more than 120 ° C, the bacteriologists are aware that by keeping the part

downstream of a membrane valve at 100 ° C, most of the risks of reinfection via the valve are already eliminated. A fortiori, if a higher temperature is maintained, the risks decrease / also at the sterilization temperature of the product, depending on the pH, the risk is it zero. AT

1 tt - 14 -! It is imperative to prevent infection of the contents of the flexible tanks and their pipes, infection originating from the gas outlet, as well as damage due to an excess of internal pressure. In addition, a slight over-! pressure is desirable at the top of flexible tanks and in flexible gas evacuation ducts.

One of the devices ensuring these three conditions is that of FIG. V. It has the advantage of being effective even in the event of a failure i; electric, i In Figure V we have: i 1: Arrival of gas from flexible tanks 15.

• 2: Gas entry into the bell 3.

3 s Gas bell or rigid tank open at its lower part.

4: Filling mass or Raschig rings ensuring maximum contact with the gas passing through this mass.

5 s Perforated bottom supporting 4.

6: Tank.

7: Antiseptic liquid.

25 8 s Pump.

9: Liquid distributor 7 on the mass of. filling 4.

10: Arrival of a filtered gas, air or a noncondensable gas as appropriate.

The interior of the devices of FIG. V can be sterilized, independently of the tanks! flexible and conduits thereof, but the case i. the most common is to sterilize all | system simultaneously.

: 35 The gases (vapor and antiseptic or mixture of non-condensable sterile gas, vapor, antiseptic) used to sterilize the flexible tanks as well as their accessories and pipes arrive from 1 to 2 and sterilize the contents of the bell 40 3, already partially disinfected by the circulation of the antiseptic liquid 7 via the pump δ Ê - 15 - and the distributor 9.

The liquid 7 heats up.

When it is certain that the sterilization is complete and that the pure steam, or the mixture of “hot sterile water mist / sterile and hot incondensable gas” has evacuated anti-residue tanks, accessories and pipes septic tanks, the hot or boiling liquid 7 is enriched with antiseptic, and the pump 8 is allowed to run.

The arrival of steam or of steam and sterile gas is interrupted upstream of 1 as well as in the liquid 7. The sheath l is closed or an aseptic valve located in 2 is closed, then air or 15 is introduced via 10 filtered neutral gas. This is completely washed and sterilized by the flow of antiseptic liquid flowing over the mass 4.

When the gases arrive in 2 the pump 8 operates continuously but the elements of the figure V 20 between 1 and 4 remain sterile even if the pump is stopped for more than a day.

It is easy to avoid a lack of liquid in the tank (6), so the only important control is the antiseptic content and the degree of soiling of the liquid 7.

The choice of antiseptic varies depending on whether the gas arriving in the bell is pure or loaded with organic matter.

Antiseptics that are not very sensitive to organic materials are, for example, formaldehyde, sulphites in a weak acid medium, halogenated derivatives of acetic acid.

Water that is acidulated and kept above 70 ° C is also suitable, but it is preferable to lower its pH to 1 by means of a non-volatile acid such as sulfuric acid and / or to add an antiseptic because aseptic must be maintained in the event of a breakdown.

Many fermentations and productions of 40 microorganisms require a development in pilot installation, whose flexible tanks have 1 -16- The agitation of the contents of the small tanks can be carried out without pump, by undergoing deformations to the walls of these tanks.

Rotary, centrifugal or metric volu-5 pumps, made aseptic, are not suitable in all cases, whether they are industrial installations and especially pilot installations.

Aseptic transfer of a closed liquid; sterile tescible, stored in a sterile flexible tank, towards a fermentation tank requires not only an aseptic metering pump, but that this does not allow microorganisms to contaminate the sterile liquid upstream of the pump.

The aseptic extraction of the deposit of organic matter or of the sediment of microorganisms from the conical bottom of a static decanter cannot allow foreign microorganisms contaminating the outlet in the open air to enter the decanter; however, a single microorganism is sufficient.

The hourly flow rates to be considered are extremely variable, the maximum being of the order of 10,000 liters and the minimum of 0.0003 liters.

This minimum corresponds to the volume of pro-25 sediment from 100 liters fermented during a year.

A new type of aseptic pump with the desired performance is shown in Figure VI. where we have 30 1: Upstream part of the rigid pump casing.

2: Actuator or motor, normally closed by the action of a spring, of the suction valve of the pinch valve type.

3: Fixed support on which the flexible sheath 35 is crushed 4 by the actuator 2.

4; Flexible sheath.

5: Port allowing the 2/3 valve to be opened from the outside.

6 s Flange or other sealed connection between 1 and 7.

40 7: Body of the rigid box.

8 s Arrival of the liquid filling the box j »_ · ______ <____ - λ __ -. - λ - 17 - as well as on the aseptic valve motors.

9: Sealed valve.

9 "i Possible reservoir for excess liquid 5 for filling the rigid box.

10: Same as 1.

11: Same as 2, but it is a discharge or discharge valve, normally closed.

10 12 s Same as 3.

13: Same as 5.

14! Rigid hose.

15: Semi-rigid pipe.

16 s Sheath.

15 17 s Jam.

18 s Same as 14.

19: Same as 15.

20: Same as 16.

21: Same as 17.

The length of the pump body 7 and the diameter of the duct 16-4-20, as well as the shape of the duct between the valves 2/3 and 11/12 depend on the flow rate per desired cycle.

An optionally sterile cylindrical sheath 25 can be put in place after opening the valves 2/3 and 11/12 through the orifices 5 and 13.

The box is sealed thanks to the stuffings 17 and 2l compressing the sheath 16-20 on the semi-rigid pipes 15-19 supported by the rigid walls 30 of the pipes 14-18.

The semi-rigid pipes 15-19 are used in stuffing and also to support the flexible sheath at the end of the rigid pipes 14-18, on the box side. They prevent the flexible sheath from being dangerously deformed.

The box being sealed and the interior of the flexible sheath being sterile, or sterilized, the box is completely filled by the piping 8 by means of cold water or another liquid 40 having a low sizing tension.

Excess liquid arrives in the tank £ - 18 - optional 9 '.

The valve 9 is closed. The pump is ready to operate as follows:

A vacuum at 8 acts on the flexible sheath at 45 and on the submerged motor of the actuator 2, which moves away from the fixed support 3. The pump sucks the liquid present at 22.

The sheath 4 being filled, the vacuum is removed at 8. The suction actuator 2 closes. When 10 a pressure at 8 is transmitted towards the interior of the box, the actuator 11 opens while the pressure increases around the sheath which applies and pushes its contents towards 23,

Valves 2/3 and 11/12 may have one or more stirrups compressing the sheath on the fixed supports 3 and 12, however, if it is necessary to avoid the presence of microorganisms downstream from the discharge valve located inside the box, it is necessary to install downstream of the packing 20 of discharge 21, at least one valve of the type represented by figure VII, where one has 23: Flexible sheath.

24: Axis of rotation.

. 25: Flexible or rigid blade, rated 24, then flexible.

25 26: Blade movement control 25.

27: Rigid support on which the sheath rests.

25 ': Position of the blade 25 when, under the action of the control 26; it completely crushes the sheath 23 against the support 27.

30 26 ': Control 26 in the low position.

This valve fitted with a spring is of the normally closed type. The liquid filling the pipe 8 and the box 1-7-10 of figure VI arrives in the hydraulic motor acting on the control 26, '35 also the valve of figure VII opens when the pressure in the box expels, via the valve 11/12, the product contained in the sheath of the box

In Figure VII 1 the crushing of the sheath, 40 between 25 and 27 expels quickly the contents thereof, so the interior of the end of the Jt - 19 - sheath does not become infected seen the frequency of purges and the fact that the flexible sheath is completely flattened over a considerable length between two parallel surfaces, that of the support 5 27 and that of the blade ·

FIG. VII is schematic and relates only to the principle of this valve, the blade 25 of which can have another profile, and obviously has a decreasing thickness of the hinge 24 at the end where acts command 26.

Safety increases by placing more than one valve type fig. VII in series, downstream of the pump and the discharge valve 11/12 can also be replaced therein by a valve type fig. VII.

15 This valve may include as a shutter, instead of the spring 25, a double control spring, therefore a control 26 and a second control • parallel to the first and acting on the second end of the blade.

Another device comprises a flexible, inflated, pressurized sheath compressor, normally crushing the sheath but rising when this relief valve must open.

In all cases, the principle remains the same:

The sheath is first crushed and therefore closed. Then the part located downstream of this sheath gradually crashes. There remains, in the flat part of the sheath, only negligrable traces of pumped product and rapid emptying of the sheath downstream, combined with deformation. rapid shedding of the sheath walls, expels outwards all portions of product likely to create a bacteriological problem, 35 so the system prevents an infection from reaching the valve 11/12 and, a fortiori , go back to valve 2/3 and worse still upstream of it.

In Figure VII, upstream and downstream of the aseptic pump there is no rigid pipe in the flexible sheath. For certain applications jé * - 20 - - If you wish to use a rigid piping downstream of the pump you can also replace at the end of this piping the valve or valves type fig.VIl with a pinch valve or ä mem-5 brane normally closed, opening and closing at the same time as valve 11/12 of> figure VI. The ideal is to have at least 2 terminal valves in series and that the compression of the sheath or of the membrane is less in the terminal valve.

'In large capacity pumps, the diameter and the length of the central box (7) of Figure VI are important and, in this location the diameter of the sheath may be close to that of the box.

In this case it may be advantageous to have in 6 and 10 a plate comprising-also a stuffing of the type 17-21.

There are then 3 boxes, i.e. 1-7 and 10.

The box 1 communicates with the vacuum source and the box 10 with the pressure source. Alternative pressure and pressure can, for example, be caused by a piston pump, the suction of which is connected to the inlet box and the discharge to the outlet box. The pressure variations in the central box are obtained thanks to internal or external pipes each comprising a flow control valve and even a spring-loaded pressure reducer.

30 This reduces the stress on the membrane by advancing the opening of the valves. When the section of the sheath is large between the valves it becomes impossible to introduce it by the first stuffing, so the sterilization of the inside of the sheath is to be carried out after mounting the pump, where the particular interest of the sheaths having a relatively constant diameter.

Partial removal of the over-board material present in the liquid upstream or downstream of the flexible tank (s) is achievable - 21 - β cooling and even assisted by the addition of a flocculent. The absence of any foreign microorganism being essential, a decanter, initially sterile, with flexible walls, having 5 or not a gas evacuation pipe, is suitable if it is equipped with an aseptic pump according to Figures VI and VII. At least the bottom of the decanter is conical or pyramidal. Its flexible walls are slowly but regularly deformed 10 in order to detach the deposits.

The arrangement of the lines for liquids and gases of the flexible tank series is shown in Figures VIII 3 'X, where there are 1: Flexible tanks 15 2: Circulation pump 3: Lines for liquid 4:' 'gas 5s ·' ” ”

The gas lines are not shown | 20 tees in Figure X, a plan view.

| When a single pump · circulates the liquid in a series of flexible tanks, it is necessary to limit the pressure drops, the effects of | these. and those of a pump stop, j 25 The diameter of the pipes can also be increased in order to allow, thanks to these pipes, the ac-! these workers and even some equipment.

This simplifies and improves the production and installation of the installations.

30 In FIGS. VIII and I} (the eventual domes improving the gas collection have not been shown. These are found on the tanks 1, FIG. VIII and on the pipes 4, FIG. IX. details, the important of which depends on the type of fermentation or culture to be carried out.

FIG. XI represents a section in a building comprising a single flexible tank, we have:

40 1 to 13 î See figure I

14 d Thermal insulation

15: Craz C sealing layer

- 22 - 16 s External construction protection.

The wall of large capacity flexible tanks facing their liquid outlet or outlet orifices is reinforced and equipped with external tensioners or cables limiting its movement. They prevent the membrane from obstructing the orifice and, if necessary, stop the filling and act on an indicator of the degree of emptying.

Another safety device stops filling before excessive tension develops in the membrane.

At least the part of the flexible tanks containing liquid is submerged.

The ratio of the heights of liquid and gas can be arbitrary. It depends on the type of fermentation or culture.

The part of the tank filled with gas can also be maintained by any mechanical means, with a view to partially stabilizing the shape of the flexible tank and reducing the low overpressure to be maintained there, including thanks to the gas sterilizer washer described in FIG.

Example 1.

22 High fermentation beer is greatly improved by refermentation in the bottle.

However, especially when it comes to relatively low alcohol beers, the risks of infection by foreign yeasts, called wild yeasts and by bacteria, especially lactic, are significant. This is all the more so since the beer, drawn off at low temperature, must be reheated to re-ferment fairly quickly.

35 This is a technology that is not very industrial and at risk.

Maturation in the bottle requires at least 15 days, better a month. In good conditions d - 23 - the quality increases for about 3 months, even longer with strong beers.

Beer re-fermented in bottles is expensive since the custody relates to the packaged product.

5 It presents significant risks and many consumers and retailers do not appreciate the presence of a deposit at the bottom of the bottles.

Also only small quantities of luxury beers are still fermented in the bottle, ίο 'The example uses must suitable for a high fermentation beer. The sterile must is aseptically cooled and aseptically stored in a submerged flexible tank (Luxembourg patent, deposit number: 85.954) 15 The sterile must is kept in motion in the aseptic storage tank forming a buffer and part of the cloudiness, or deposit, in suspension is aseptically removed during its aseptic transfer to the main fermentation flexible tank containing a high, pure fermentation yeast, introduced aseptically during the start-up of the installation.

Fermentation, between 15 and 30 ° C, lasts 3 to 7 days, with agitation carried out aseptically.

25 The beer, continuously fermented, contains a maximum of 0.595 to 1.38% of unfermented wort, so either sterile wort or sterile sugar syrup is introduced aseptically during transfer to the submerged flexible container (s) 30 secondary fermentation. Excess yeast, mainly from clumps and large cells, is separated from this stream of young beer by means of a centrifugal separator adjusted accordingly.

Secondary fermentation, carried out under agitation; · 35 tion, lasts 15 to 60 days at a temperature between 15 and 30 ° C.

The ripe, flat beer is then treated., According to well-known rules in the brewery, by centrifugation or filtration, saturation, placing λ 40 in barrels, bottles or cans.

As secondary fermentation allows

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The bility of beer is rather higher than that of tall, non-referenced beers.

Maintaining the particular taste and fragrance characteristics due to resealing requires that the packaged beer contain very little oxygen. This is achieved thanks to the new commercial filler (less than 0.1 ppm of oxygen after packaging)

The wort production being X liters / day 10 the capacity of the main fermentation tank (s) varies between 3 X and 7 X and that of the secondary fermentation group from 15 X to 50 X, so it suffices to proportion the capacity and / or the number of flexible tanks and their 15 additional equipment to cover all needs from that of a pilot installation to that of a very large brewery.

Example 2.

It is a beer of low fermentation. The main fermentation, slower, rather takes place below 10 ° C and the secondary fermentation between 1 and 5 ° C.

Continuous fermentation is not yet suitable, so it is carried out in cylin-25 dro - conical tanks without seeking to achieve saturation with the carbonic acid of fermentation.

The beer would be salable but it is refined by continuous refermentation. This requires degassing the i> rst, filtering it on diatoms or centering it and then pasteurizing it. The sterile and cooled flat beer is then aseptically introduced into the refermentation group. The latter and the following operations are similar, except for the temperature, to what was described in Example 1.

The use of an aseptic buffer tank is justified or not, depending on the frequency of emptying of the main fermentation tanks.

Example 3.

40 The data are those of Example 1, but

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- 25 - pasteurization and cooling, a fermentation in a flexible tank using a selected lactic acid bacteria.

The proportions of products treated with a yeast or with a lactic acid depend on the type of beer desired and also on the amount of extract fermentable by lactic acid bacteria.

The sour beer is filtered, pasteurized, cooled, and then aseptically mixed with the "ripened" beer.

The mixture undergoes a maturation - complementary improving especially the colloidal stability.

The stabilized mixture is then conditioned according to the rules of the art as in Example 1.

Example 4.

A pilot plant is designed to rapidly develop a fermentation process. This installation uses numerous flexible tanks, the unit capacity of which is less than one cubic meter. In addition to the apparatuses and devices of the types I to XI, the installation comprises a large flexible reservoir 25 in which the stock of sterile and refrigerated liquid, to be fermented, is stored away from any oxidation, aseptic accessories such as static decanters, small hydro-cyclones, sterilizable and removable filters 30 and continuous mini-pasteurizers.

The contents of certain flexible tanks are agitated by pumping. That of other reservoirs is only agitated by mechanical actions deforming the membrane of the reservoir and causing the liquid to circulate.

Each flexible tank is immersed in aerated water. Water is used to control the temperature and that of the oxidation of the products via flexible tanks, 40 piping and accessories.

Microorganism, durations and flow rates, microorganism content, mixtures of products having been fermented by different microorganisms, elimination of an excess of microorganisms, decantation, centrifugation, filtration.

The tests are carried out in parallel, s each test using tanks in series.

The generally poor results obtained in conventional pilot plants using rigid fermentation tanks and the very high cost of this conventional equipment necessitate recourse to the process which is the subject of the present invention. dice may be suitable for installation on an industrial scale.

!

Claims (15)

1. Flexible tanks, "reactors" or "fermenters", are constructed from flexible membranes of thermoplastic materials or elastomers, reinforced or not by a textile and can be sterilized in the factory by known means before setting up or sterilized after setting up and before putting into service, all the parts of the equipment which could be contaminated being sterilizable and in particular the connections / 25 with the installations for supplying raw materials and those for withdrawing products obtained. 1. The system implements an initially sterile apparatus, comprising at least one flexible reservoir immersed in a liquid of suitable characteristics, at least up to the upper level of the product which it contains; a liquid kept in motion where. takes place either a fermentation or a culture in the absence of any undesirable foreign microorganism, the flexible pipes and other possible devices with flexible walls containing a liquid, being submerged, while the possible devices and pipes with flexible walls, containing a gas, are not. These lines can float on the surface of the immersion liquid. 2. The reactors according to B1 are immersed in a liquid, preferably water possibly added with salts or products in suspension - in order to regulate its density, in order to reduce the mechanical stress on the membrane as well as to regulate the temperature inside the reactor. 2. The system which is the subject of claim A1 comprises several tanks arranged in series. 25 3 - The systems which are the subject of claims Al and A2 comprise one or more decan-tors with flexible walls, initially sterile,. Immersed like the flexible tank or tanks. 3. There are disposed in the basin and in the immediate vicinity of the reactors nozzles directing towards the walls thereof pulsed immersion liquid in order to temporarily deform the flexible walls and thereby detach the deposit tending to s '' train there despite the movement maintained / in . mid - 29 - 4. The surface of the immersion liquid of the apparatus of which question in B2 can be covered by a floating film in order to reduce evaporation, caloric transfers or transfers of gaseous products (oxygen for example). 4. The immersion liquid is in one or 30. several tanks in concrete, masonry, metal, plastic or earth and dikes possibly made waterproof thanks to a plastic film. 5. The whole apparatus or, at the very least, the reactors and the tanks containing the immersion liquid are placed in a room whose atmosphere has a controlled composition. 10 6 - The membranes from which the apparatus is constructed are chosen so as to limit the supply of oxygen to the mixtures during fermentation or storage. 5. The immersion liquid is cooled or heated. 35 6 - The immersion liquid is deaerated by physical, chemical or biological process. 7. The composition of these membranes is chosen so as to avoid migration to the liquid during the preparation of any noxious materials and the permeation of the flavors towards the outside. 7. The immersion liquid is water or a mixture of • dissolved and suspended water and salts. increasing its density and / or viscosity. 2, ”. t - 28 - 8. The membranes consist, as a consequence of the requirements expressed in 7, of one or more of the following materials: - Low, medium or high density polyethylene. - Polyethylene ionomers. - Polypropylene and its copolymers. - Polyvinyl chloride and its copolymers, plasticized or not. - Polyvinylidene chloride and its copolymers. - Polyvinylidene fluoride and its copolymers. - Ethylene-vinyl alcohol copolymers as such or modified. 30. Polyamides. “- Linear polyesters. - Cellulose derivatives. - Metals in thin sheets like aluminum. These materials are used in the form of films, either unstretched, or stretched monoaxially, or biaxially. If the mechanical constraints require it, the membranes will include a textile reinforcement based on natural or synthetic fibers including glass. 40 Pnnr 1 oc mQTnhranûC pnmnl ûvqc (miilf ιηηηην, ΰο λ £ - 30 - 5 the adhesion between the various constituents is obtained by thermal welding or by means of adhesives or "compatibilizers". The membranes used can also be fabrics waterproofed by means of elastomer-10 res, polyisobutylene for example. 8. Transfers of liquids or gases with the outside, upstream and downstream of the flexible tanks, are carried out by means of devices preventing any introduction of undesirable microorganisms into pure culture or even any contamination by the latter of the content. of the possible flexible tank and aseptic storage of the sterile liquid to be treated. 9. The membranes are assembled to form the "reactors" of the pipes and fittings between reactors and flexible pipes or between flexible pipes and rigid plastic or metal pipes. They are assembled by welding or gluing, if necessary, by seams made waterproof by coating, gluing or welding. 9. A slight, constant overpressure is maintained * · 10 in the gas phase of the flexible tanks and the pipes evacuating the gases from them. ël Apparatus allowing the system to operate characterized by the following points, 15 separately or in combinations s 10. The circulation and homogenization of the liquids contained in the apparatus is preferably carried out by means of circulation pumps, this does not exclude the use of conventional agitators or that of deformations brought to the flexible membranes. 11. All fittings and sealing devices for rotating shafts and other places which could, in the short or long term, allow the introduction into the apparatus of microorganisms or their spores, are immersed in a liquid. antiseptic or are placed in hermetically sealed and sterilizable containers * 30, the immersion in an antiseptic being if necessary preceded by intensive watering with this antiseptic if it is essential to prevent any re-infection during the short period of time required to immerse the apparatus presenting risks of reinfection by micro-leaks and having just been sterilized. 12. A gas washer, sterilized, in which the gases pass through a contact mass sprayed in a circuit closed with a cold solution and antiseptic or kept at least 70 ° C and also antiseptic, before escaping by bubbling in the antiseptic liquid now a constant overpressure in the gas washer protects the interior of the gas pipes against any risk of infection by undesirable microorganisms, avoids excessive internal pressures and maintains in Jas flexible pipes carrying the gases 5 as well as in the part of the reactors filled with gas an adjustable and constant overpressure. 13. Aseptic pump, suction and treading, sterilizable, of adjustable dimensions and unit capacity making it possible to avoid any passage of microorganisms-10. U ”downstream upstream, constituted by an internally sterile flexible sheath before its introduction into the pump body or even sterilizable after its installation, a box and at least three aseptic valves, of the sleeve valve type, normally! closed by springs, at least two of which, one for suction and the other for delivery, are located in the box, and at least one, for delivery, outside the box. ! 20 The rigid box has two and better three sections. The first contains an orifice with a sheath inlet stuffing, at least one suction valve and at least one orifice which can be closed by plug enabling the corresponding valve to be opened from the outside, in order to introduce the sheath. The second section of the box constitutes the chamber of variable dimensions where the sheath is located, of constant diameter or not between the suction valve (s) and the discharge valves, sheath 30 whose deformations carry out the transfers of liquids or gases. This central box can also include the arrival of the working fluid, a liquid with low vapor pressure such as cold water, as well as an overflow orifice provided with a valve and 35 connected if necessary to a reservoir receiving the excess motor fluid. The third section of the box is identical to the first section but contains at least one discharge valve. The jams compress the wall of the flexible sheath 40 on a semi-rigid inner pipe in the casing and prevents, by gradually deforming, that the sheath is too deformed in the vicinity of the rigid pipe when it becomes flat. The flu-ide whose pressure modifications 5 act, in the box, on the sheath and the motors of the · valves actuates, by means of piping, the motor or motors of the discharge valve (s) located downstream of the box, the der-. At least three of these valves being of a new type, self-cleaning (as described in the text of the patent, page 18-19 and FIG. VII) characterized by the action of the sheath shutter. This first crushes the sheath, then simultaneously increases the flattening pressure as well as the length of the flattened zone downstream, zone completely empty of liquid. 14. The tension of the springs, keeping the valves normally closed, is adjusted so that a discharge valve becomes the safety valve of the mini-tank from which it discharges. 15. The flexible sheath of the pumped liquid transfer can be externally protected by an independent plastic, elastomer or textile sheath, the role of which is purely mechanical. I IS