Process for the production of non-alcoholic beer and device for effecting this process

Abstract: The invention relates to a process for the production of non-alcoholic beer, the pH-value of the wort being adjusted in that wort is fed through a bioreactor with lactic acid producing bacteria immobilized on a specific carrier, the acidified wort flowing out of the bioreactor being allowed to reside in a post-treatment container and subsequently being mixed with a non-acidified wort which is to be adjusted to a specific pH-value. A preferred embodiment of the carrier includes microfibres or microparticles of diethyl-aminoethylene-substituted cellulose (DEAE-cellulose), which are bonded and agglomerated to one another by polystyrene. The invention also includes an apparatus for effecting this process. The large-scale acidification of wort by means of immobilized lactic acid producing bacteria is made possible by means of this process.

Description: The invention relates to a process for the production of non-alcoholic beer and a device for this. The term "non-alcoholic beer" refers to different products in different countries. Thus, for example, in Spain, a product with an alcohol content of less than 1 vol.-% is viewed as non-alcoholic, while the limit in Holland for this is 0.5 vol.-%. In the Arabic States, the alcohol content of non-alcoholic beer may not exceed 0.05 vol.-%. In the USA and some other states, a drink with an alcohol content of less than 0.5 vol.-% may not be labelled as being beer. In terms of the invention, the term "non-alcoholic" is to be understood as implying that a beer is meant which has a greatly reduced alcohol content with respect to normal alcoholic beers. A common way of producing non-alcoholic beer consists in removing the alcohol from normal alcohol-containing beer. However, the processes for this are very costly and the applied process steps such as distillation and the membrane separation technique also lead to the removal of aroma components and therefore to an often watery tasting final product. A further manner of procedure for producing non-alcoholic beer uses the contacting of the wort with active yeast under specified conditions. This process is commonly effected in batch reactors and most recently also with continuously operating bioreactors which are packed with immobilized yeast (European patent application EP 0 424 794 A2). The process parameters, in particular the temperature in the bioreactor and the flow velocity through the bioreactor, are selected such that the alcohol formation is suppressed but that other metabolic reactions with yeasts can occur which lead to the formation of important flavour components and the reduction of off-flavours. The wort used for the process in this case is produced in the usual manner adapted for preparing non-alcoholic beer. In comparison to normal alcoholic beers, non-alcoholic beer is more sensitive to microbiological spoiling during the production process and also during storage. In order to avoid this risk, the pH-value is usually already reduced in the wort, so that the stability of the product is increased. The simplest manner of doing this consists of adding an acid to the wort, lactic acid being preferred for reasons of flavour. Legal regulations in certain countries forbid the addition of acid in the process for beer production if the acid is not formed on a natural basis by fermentation during the process. In this case, one often then proceeds by adding lactic acid forming bacteria to wort in a batch fermentor, and the wort treated in this manner is mixed in specified ratios with the wort of which the pH-value is to be adjusted. However, the process carried out according to a batch operation for acidifying the wort includes the disadvantage that the growth and activity of the lactic acid producing bacteria can only be controlled with difficulty so that the quantity of produced lactic acid and, therefore, the pH-value of the treated wort cannot be precisely adjusted. Furthermore, the speed of the lactic acid production in the batch reactor is relatively slow so that, especially in big breweries where many brews are made each day, a large biomass must be applied in order to produce enough lactic acid and to obtain the suitable pH-value. On the whole, acidification according to the above mentioned batch operation process is very time consuming and often requires very large facilities. The immobilization of various lactic acid producing bacteria is known in the state of the art, the immobilization being effected by means of various techniques for trapping or encapsulating the bacteria in carrier materials, as described, for example, in "Enzymes and Immobilized Cells in Biotechnology", Benjamin/Cummings Publishing Compl., Inc., 1985, published by Allan I. Leskin. However, these techniques have considerable disadvantages so that a transformation to the industrial scale has not been satisfactory up to now. The applied carrier materials, for example alginate pearls, are comparatively soft and cause unnecessary high pressure drops in packed bed reactors when high flow velocities or large units are used. Furthermore, in the known processes according to the state of the art, the immobilization is not carried out in the bioreactor itself, but in a separate facility when the carrier particles are brought together with the bacteria. Finally, the carrier material cannot be reused when the system is contaminated or interrupted for other reasons. The above described disadvantages led to the fact that the application of immobilized lactic acid producing bacteria for acidification in large scale beer production was not viewed as promising success. It is an object of the present invention to provide a process according to which the immobilized lactic acid producing bacteria for acidification of wort can be applied and the above-mentioned disadvantages simultaneously avoided. A further object consists in providing a corresponding apparatus for effecting such a process. The above first-mentioned object is solved by a process for the production of non-alcoholic beer, in which the pH-value of the wort is adjusted with lactic acid producing bacteria, which is characterized in that a) a wort which is free or substantially free from hops or hops extract is allowed to flow through a bioreactor which contains immobilized lactic acid producing bacteria on the surfaces of a substantially non-compressible carrier which consists of a continuous, porous matrix or of dimpled or reticular, porous particles, the matrix or the particles having a structure which is formed by a loosely associated plurality of microparticles or microfibres which are chemically, adhesively or mechanically interconnected at least at some contact points between the individual microparticles or microfibres, the microparticles or microfibres being composed of a material, in particular a resin, with anion exchanger properties; b) the acidified wort obtained from the bioreactor is allowed to reside in a post-treatment container; c) the acidified wort from the post-treatment container treated in this manner is mixed with non-acidified wort to be adjusted to a specified pH-value; and d) the mixture is then further processed into non-alcoholic beer in a known manner. The wort used in step a) contains no or substantially no hops or hops extract since the lactic acid producing bacteria are predominantly sensitive with respect to substances contained in the hops and are killed off. Preferably, the wort used in step a) is pretreated in such a way that it is freed of insoluble components and preferably boiled and/or stripped with nitrogen, carbon dioxide or steam bubbles and/or clarified. The boiling is usually carried out at a temperature of 100-105°C, whereas the stripping generally occurs at 99-102°C. The boiling or stripping steps are especially performed in order to eliminate impurities of DMS and/or DMSO. The clarification is performed at a preferred temperature of 70 to 80°C, wherein a temperature of 75°C is especially preferred. The wort is preferably adapted to a temperature of 47 to 52°C before it is placed in the bioreactor. Particularly preferred, the wort is adapted to 48°C. After adapting the temperature and before introducing the wort in the bioreactor, carbon dioxide may be percolated through the wort or directly added to the wort stream, preferably up to the saturation of the wort. Thereby a carbon source is introduced in the wort being useful for metabolic activities. Furthermore, the pH-value is lowered. According to a preferred embodiment, the wort is allowed to flow through the bioreactor in step a) at a temperature of 47 to 52°C, at an overpressure of 0 to 5 bars, and with a contact time of 5 to 60 minutes. Particularly preferred process parameters are a temperature of 48°C, an overpressure of 0 to 2 bars, and a contact time of 20 minutes. The carrier material used for the immobilization in the reactor consists preferably of a resin which is derived from cellulose or rayon and modified chemically in such a manner that it has anion exchanger properties. A particularly preferred embodiment includes microfibres or microparticles of diethyl-aminoethylene-substituted cellulose (DEAE-cellulose), the microparticles or microfibres being adhesively connected by agglomeration with polystyrene. Hydrophobic polymers other than polystyrene can be suitable for the agglomeration, as for example, melamine-formaldehyde resins and epoxy resins. According to a further embodiment, the carrier material used for the immobilization of the lactic acid producing bacteria can consist of porous, sintered glass or ceramic materials. Homofermentative bacteria are preferably used as lactic acid producing bacteria which mainly produce lactic acid and some flavour components which positively influence the aroma of the beer, and which produce the lactic acid in a constant and sufficient quantity. In this case, bacteria of the strains Lactobacillus amylovorus, plantarum and helveticus can be mentioned as examples. The bacteria can be immobilized on the carrier by initially sterilizing the carrier packed in the bioreactor before the immobilization with hot, caustic soda solution, neutralising and rinsing with water and then pumping a separately cultured bacteria suspension through the bed carrier in order to bind the bacteria on the surface of the carrier. The loading quantity of bacteria on the carrier lies at approximately 10 8 to 10 12 bacteria cells/g dry carrier. The quantity of loaded carriers in the bioreactor and the process parameters during the through-flow of the wort through the reactor are preferably adjusted such that the lactic acid concentration of the outflow from the bio-reactor is preferably 2 to 6 g/l, a value of 5 g/l being particularly favourable. The pH-value of the outflow in this case lies favourably in the region of 3.3 to 4.0, wherein a value of 3.8 is particularly advantageous. The lactic acid producing bacteria applied on the carrier are preferably renewed after several months in that an aqueous caustic soda solution (2 wt.%) at a temperature of 60 to 80 °C is allowed to flow for a few hours through the reactor, and subsequently sterilising, neutralising, washing and treating this with a separately produced bacteria suspension as described above is effected. As a dying off of the mentioned bacteria can ensue with a very high loading of the carrier with the lactic acid producing bacteria and, therefore, with a high concentration of lactic acid produced in the reactor, it is useful not to select too high a concentration of the bacteria in the reactor, and to instead effect a post-fermentation in accordance with step b) of the inventive process. In this case, the wort is transferred into a post-treatment container after treatment in the bio-reactor. Simultaneously with the wort, small quantities of bacteria in the reactor absorbed on the surfaces of the carrier are transferred with the outflow into the post-treatment container. These bacteria effect a post-fermentation in the container, i.e. the lactic acid concentration of the wort increases in the post-treatment container and the pH-value drops further. According to a preferred embodiment of step b), the residing time in the post-treatment container is set such that the concentration of the lactic acid amounts to 9 to 15 g/l, and in particular preferably 10 g/l. In this case, the pH-value is favourably adjusted to a range of 2.8 to 3.2, the optimum pH-value being <3. In step c) of the inventive process, the acidified wort removed from the post-treatment container is mixed with non-acidified wort. The mixing ratio is preferably selected in such a way that the resulting pH-value lies in the range of 4.0 to 4.5, and more preferably 4.2, in the case where the wort is treated in step d) in a bioreactor with immobilized yeast. When the wort is treated with non-immobilized yeast according to step d), the preferred pH-value after the mixing in step c) lies within 5.0 to 5.5. The wort to be adapted to a specific pH value and to be mixed in in step c) is preferably pretreated by removing insoluble components, boiling and/or stripping (with nitrogen, carbon dioxide or steam bubbles) and/or clarification as indicated previously for the wort to be acidified by immobilised lactic acid producing bacteria. According to step d) of the inventive process, the wort adjusted to a specified pH-value is then preferably cooled to a temperature of 0°C to 2°C, filtered and subsequently treated with yeast. Preferably, the treatment with yeast ensues in a bioreactor with immobilized yeast. In a particularly favourable embodiment, the carrier for the yeast in this case is of the same type as in the immobilization of the lactic acid producing bacteria, i.e. a continuously porous matrix or dimpled or reticular, porous particles, the matrix or the particles having a structure which consists of a loosely associated plurality of microparticles or microfibres which chemically, adhesively or mechanically interconnect at least at some contact points between the individual microparticles or microfibres, the microparticles or microfibres being formed of a material, preferably a resin, which has anion exchanger properties. Preferably, the carrier is formed of a resin which is derived from cellulose or rayon and chemically modified in order to have anion exchanger properties. A most particularly preferred embodiment includes microfibres or microparticles of diethylaminoethylene-substituted cellulose (DEAE-cellulose) which are bonded or agglomerated with one another by polystyrene. A further embodiment contains as carrier for yeast porous, sintered glass or ceramic materials. According to a particularly preferred embodiment of process step d), the bioreactor can include several columns, advantageously placed under pressure and/or connected in parallel. The second aspect of the above-mentioned object is solved by an apparatus for the production of non-alcoholic beer, in particular for the acidification of wort by means of lactic acid producing bacteria, which is described in more detail in the enclosed figure. Figure: Schematic flow diagram of an apparatus for the production of non-alcoholic beer. The apparatus includes a container for the manufacture of mash (1); a container (2) for removing insoluble components; a boiling container (3) and/or a stripping device (4) and/or a whirlpool (5) for clarification; heat exchange equipment (6); a device (7) for introducing CO 2 into the wort; a bioreactor (8) packed with a loaded carrier of the above-mentioned type for the acidification; a post-treatment container (9) and a temperature control device (10) for the temperature in the post-treatment container (9); means (11) for introducing the acidified wort from the container (9) into one or more of the devices (12) to (17) explained below; a mash container (12); a container (13) for removing insoluble components; a boiling container (14) and/or a stripping device (15) and/or a whirlpool for clarification (16); a mixer (17); a cooling device (18) for the mixed wort; a filtration unit (19); a holding tank (20); and a bioreactor with immobilized or non-immobilized yeast (21). According to a preferred embodiment, the bioreactor (21) consists of several columns, preferably connected in parallel and/or placed under pressure. The above described process for the production of non-alcoholic beer according to which lactic acid for acidifying the wort is continuously produced by means of immobilized lactic acid producing bacteria includes the advantage that, on account of the binding of the bacteria to the surface of the carrier, high and stable cell concentrations can be achieved so that the lactic acid can be produced in a sufficiently high quantity in a short period of time under controlled conditions. The continuous process makes possible short residence times of the worts in the reactor and also results in smaller dimensions of the reactor in comparison to batch operations. The treated product is contaminated only with small quantities of biomass and does not require any work-intensive separating operations. The carrier especially applied in the claimed process can be repeatedly used and has a long lifetime. Finally, no unnecessary pressure drops are generated in the reactor due to the substantially non-compressible carrier. The bioreactor according to the invention includes a high capacity and a high flexibility with regard to the production of lactic acid. The bioreactor can be kept for several weeks under stand-by conditions and can be started up very easily. The processes in the bioreactor can be performed completely automatically and in a closed system, so that a good biological condition is obtained with no contamination by foreign bacteria or yeast cells. The inventive bioreactor enables a compact installation and the saving of energy and costs. The present invention is described in more detail with respect to the following examples. Example 1: Production of a carrier 25 parts cellulose were mixed with 25 parts aluminum oxide and the mixture was compounded with 50 parts polystyrene for approximately 10 minutes on a two-roll mill heated to 180 to 200°C. After cooling, the composite body was ground and pulverized to a particle size of 0.420 to 0.149 mm. 220 g of the pulverized composite body were reduced to a slurry in 616 ml water containing 176 g Na 2 SO 4 and 26.4 g NaOH. The slurry was heated to 40°C and then 57.2 g of a 5% aqueous solution of diethyl aminoethyl chloride x hydrochloride (DEC) were added to the slurry with mixing at a rate of 0.7 ml/min in a period of 1 hour. Subsequently, 26.4 g NaOH dissolved in 26 ml water was added to the slurry and then a further 27.25 g of DEC-solution at a rate of 0.7 ml/min. The temperature of the slurry was raised to 60°C and this temperature was maintained for 15 minutes. A water quantity which was approximately the same as the volume of the slurry was added and the composite body was obtained on a sieve with a low mesh size of 0.250 mm. The composite body was washed on the sieve with water and then again reduced to a slurry in a quantity of water which corresponds to the previously added quantity. This slurry was adjusted to a pH-value of approximately 7 with HCl, dewatered on filter paper and dried. Example 2: Immobilization of the bacteria on the carrier 10 g of granular DEAE-cellulose of example 1 were reduced to a slurry in distilled water and soaked for 5 hours with occasional stirring. The hydrated carrier was then decanted with distilled water and transferred to a glass column with an inner diameter of 15 mm, in which it formed a bed with a height of 145 mm. Lactic acid producing bacteria of the type Lactobacillus amylovorus were cultured in wort with no hops or hops extract for 48 hours at 46°C. 50 ml of the cell suspension were pumped through the carrier bed at a flow velocity of 25 ml/h and, subsequently, a further 50 ml of distilled water were pumped through. The outflow of the column was collected until it was clear, the total volume amounting to 75 ml. Example 3: Treatment of wort with immobilized lactic acid producing bacteria Wort was fed through the bioreactor of the type of example 2 including a diameter of 90 cm and a bed-depth of 40 cm at a temperature of 48°C, an overpressure of 0.5 bar and a contact time of 20 minutes. The sugar content of the fed wort amounted to 15.1 % according to the refractrometer scale of Brix, and the pH-value amounted to 5.50. The pH-value of the column outflow was 3.85 and the concentration of lactic acid was 6 g/l.

Claims: Process for the production of non-alcoholic beer, wherein the pH value of the wort is adjusted with lactic acid producing bacteria, characterized in that a) a wort which is free or substantially free from hops and hops extract is allowed to flow through a bioreactor, which contains immobilized, lactic acid producing bacteria on the surfaces of a substantially non-compressible carrier, which is composed of a continuous, porous matrix or of dimpled or reticular, porous particles, wherein the matrix or the particles have a structure which consists of a loosely associated plurality of microparticles or microfibers, which are interconnected chemically, adhesively or mechanically on at least some contact points between the individual microparticles or microfibers, said microparticles or microfibers consisting of a material, preferably a resin, which has anion exchanger properties; b) the acidified wort which is obtained from the bioreactor is left in a post-treatment container; c) the thus treated acidified wort from the post-treatment container is mixed with non-acidified wort which is to be adjusted to a predetermined pH value; and d) the mixture is then further processed in a manner known per se to make non-alcoholic beer. A process as in claim 1, characterized in that the wort being used in step a) is pretreated by removing insoluble components, and preferably also by boiling and/or stripping with nitrogen, carbon dioxide or steam bubbles and/or clarifying. A process as in claim 1 or 2, characterized in that the wort is adapted to a temperature of 47-52°C before it is transferred to the bioreactor. A process as in claim 3, characterized in that the wort is contacted with carbon dioxide after adapting the temperature and before introduction in the bioreactor. A process as in one or more of the preceding claims, characterized in that in step a) the wort is caused to flow through the reactor at a temperature from 47 to 52°C, at an overpressure from 0 to 5 bars and with a contact time from 5 to 60 minutes. A process as in one or more of the preceding claims, characterized in that the carrier is a resin which is derived from cellulose or rayon and is chemically modified to give the resin anion exchanger properties. A process as in claim 6, characterized in that the carrier consists of microfibers or microparticles of diethylaminoethylene-substituted cellulose (DEAE cellulose) which are agglomerated to each other by polystyrene. A process as in one or more of the preceding claims, characterized in that the bacteria producing the lactic acid are homofermentatively fermenting bacteria. A process as in one or more of the preceding claims, characterized in that the bacteria are immobilized on the carrier by initially sterilizing the carrier packed in the bioreactor with hot, caustic soda solution, neutralising and rinsing with water and then allowing a separately cultured bacteria suspension to flow through the reactor bed. A process as in one or more of the preceding claims, characterized in that the loading capacity of the carrier is 10 8 to 10 12 bacteria cells/g dry carrier. A process as in one or more of the preceding claims, characterized in that the lactic acid concentration in the outflow of the bioreactor is from 2 to 6 g/l. A process as in one or more of the preceding claims, characterized in that the pH value of the outflow of the bioreactor is in the range from 3.3 to 4.0. A process as in one or more of the preceding claims, characterized in that the bacteria are regenerated after several months by rinsing the bioreactor with a caustic soda solution (2 wt.%) at 60 to 80°C for a few hours and by subsequent sterilizing, neutralizing, rinsing and loading of the carrier with a bacteria suspension. A process as in one or more of the preceding claims, characterized in that the residence time in the post-treatment container is adjusted in step b), so that the lactic acid concentration is from 9 to 15 g/l and the pH value is in the range from 2.8 to 3.2. A process as in one or more of the preceding claims, characterized in that the acidified wort derived from the post-treatment container is mixed with non-acidified wort to be adjusted to a predetermined pH value in step c) in a mix ratio, such that the pH value of the mixture is in the range from 4.0 to 4.5 and that the mixed wort in step d) is treated in a bioreactor with immobilized yeast. A process as in one or more of the preceding claims 1 to 14, characterized in that the acidified wort derived from the post-treatment container is mixed with non-acidified wort to be adjusted to a predetermined pH value in step c) in a mix ratio, such that the pH value of the mixture is in the range from 5.0 to 5.5 and that the mixed wort in step d) is treated in a bioreactor with non-immobilized yeast. A process as in one or more of the preceding claims, characterized in that the non-acidified wort to be set to a predetermined pH value is freed of insoluble components, boiled and/or stripped with nitrogen, carbon dioxide or steam bubbles and/or clarified before mixing with the acidified wort. A process as in one or more of the preceding claims, characterized in that after the mixing performed in step c), the wort is cooled to a temperature of 0 to 2°C, filtered and treated with yeast. A process as in one or more of the preceding claims 15, 17 and 18, characterized in that the yeast is immobilized on the surface of a substantially non-compressible carrier, which consists of a continuous porous matrix or of dimpled or reticular porous particles, wherein the matrix or the particles have a structure which consists of a loosely associated plurality of microparticles or microfibers, which are connected chemically, adhesively or mechanically at least on some contact points between the individual microparticles or microfibers, the microparticles or microfibers consisting of a material, preferably a resin, with anion exchanger properties. A process as in claim 19, characterized in that the carrier is a resin which is derived from cellulose or rayon and is chemically modified to have anion exchanger properties. A process as in claim 20, characterized in that the carrier consists of microfibers or microparticles of diethylaminoethylene-substituted cellulose (DEAE cellulose) which are agglomerated with polystyrene. Apparatus for performing the process as in one of the claims 1 to 21, characterized in that it comprises: a container for the production of mash (1); a container (2) for removing insoluble components; a boiling container (3) and/or stripping device (4) and/or a whirlpool (5) for clarification; a heat exchanger (6); a device (7) for introducing CO 2 into the wort; a bioreactor (8) with lactic acid producing bacteria immobilized on the surfaces of a carrier as in claim 1; a post-treatment container (9); a temperature control apparatus (10) for adjusting the temperature in the post-treatment container (9); means (11) for introducing the acidified wort from the post-treatment container (9) into one or more of the following devices (12) to (17); a container for the production of mash (12); a container (13) for removing insoluble components; a boiling container and/or a stripping device (15) and/or a whirlpool (16) for clarification; a mixer (17); a cooling device (18) for the mixture of the acidified wort and the non-acidified wort; a filtration unit (19); a holding tank (20); and a bioreactor (21) with immobilized or non-immobilized yeast. Apparatus as in claim 22, characterized in that the bioreactor (21) comprises a plurality of columns, preferably under pressure and/or parallel to each other.

Inventors: BACK WERNER PROF DR, LOMMI HEIKKI, SWINKELS WILHELMUS JACOBUS PET, VILJAVA TIMO TAPIO, BACK, WERNER, PROF. DR., LOMMI, HEIKKI, SWINKELS, WILHELMUS JACOBUS PETRUS MARIA, VILJAVA, TIMO TAPIO
Applicants: CULTOR OY, CULTOR LTD.
Application year: 1993
Main classification: C12C12
IPC codes: C12C11, C12C13, C12C12, C12N11, C12G3, C12C7