WO2010128117A1

WO2010128117A1 - Method and composition to reduce unfiltrable haze in fermented beverage

Info

Publication number: WO2010128117A1
Application number: PCT/EP2010/056206
Authority: WO
Inventors: Pablo Alvarez
Original assignee: Cbs S.P.R.L
Priority date: 2009-05-06
Filing date: 2010-05-06
Publication date: 2010-11-11

Abstract

A method to reduce haze in a beverage by adding a sufficient amount of an enzyme composition comprising an alphaglucanase.

Description

Method and composition to reduce unfiltrable haze in fermented beverage

Field of the invention [0001] The present invention relates to a new method and a composition to reduce (unfiltrable) haze in fermented beverages, especially in beer, wine and cider by adding a sufficient amount of an enzyme or an enzyme mixture composition .

Background of the invention and state of the art

[0002] Haze is a well-known phenomenon in the beverage industry, including beer, wine and cider industry. [0003] Several methods were developed to reduce haze in beverages, because haze negatively affects the customers' perception of beer, wine or cider. [0004] As mentioned in Malcorps et al (Glycogen released by the yeast as a cause of unfilterable haze in the beer, 2001, Technical Quarterly, Vol. 95, pp 95-98), many types of haze may occur in beer. As haze consists of small particles suspension in a beverage (beer, wine, cider) , monitoring of haze is usually conducted using a low (e.g. 25°) scattering angle. Prediction of beer colloidal stability is performed by a measure with a scattering angle of 90° following recommendations of EBC (European Brewery Convention) , ASBC (American Society of Brewing Chemists) and MEBAK (Mitteleuropaische Brautechnische Analysen- kommission) [0005] The most common haze is formed upon chilling of the beverage and is a result of interaction between phenolic and proteic compounds.

[0006] A first method that was developed to reduce (common) haze consists of a chilling of the beverage (preferably under 0⁰C) followed by a filtration at this temperature of this (common) haze. This method is well known by the person of art as "chill proofing". [0007] Alternatively, polyvinylpolypyrrolidone (PVPP) and/or egg-white can bind phenolic compound of (common) haze and allows a filtration of this (common) haze .

[0008] Another approach consists into an addition of proteases, such as papain or of proteases presenting a specific activity to reduce (common) haze. However, proteases should be dosed carefully as proteins are necessary to form stable beer foam.

[0009] EP 1 326 957 Bl describes a reduction of (common) haze in a beverage after addition of a prolyl- specific endoprotease (sold under the name of Clarex; DSM) to this beverage.

[0010] Proteins can also be eliminated by silica gel or gallic tannins addition by forming precipitates which can be filtered. These treatments can also have a negative impact on beer foam stability.

[0011] However, there is a tendency to increase wort gravity (high gravity) , temperature and volumes during the fermentation step on an industrial scale to boost productivity. [0012] In these new industrial processes (wherein original gravity and/or temperature and/or volumes are increased) another haze is formed and persists, even after use of one of the above-mentioned methods to reduce

(common) haze (upon a "chill proofing" of the beverage, such as the filtration after addition of PVPP or silicagel or tannins or proteases) .

[0013] This persistent haze called "unfilterable haze" (Malcorps et al . , 2001) is thought to be made of different compounds than of (common) haze (e.g. alpha- glucans, beta-glucans, glycoproteins, pentosanes) and is resistant to the above-mentioned classical palliative treatments, like the addition of proteases and/or fining agents . [0014] Measurement at about 90° scattering angle is known to better evidence smaller particles (Malcorps et al . , 2001) . The unfilterable haze is thought to be made of very fine (i.e. unfilterable) particles. [0015] Malcorps et al . highlighted the polysaccharidic nature of this "unfiltrable haze". However, these authors did not find an enzymatic treatment solution to this problem, with the requisites of working at 0-5⁰C, being quickly deactivated by beer pasteurisation and without changing the organoleptic qualities of the final product.

[0016] Recently, it was discussed that an addition of (the above-mentioned) proline-specific protease sold under the name Clarex (DSM) during beer fermentation, while reducing the level of common (chill) haze as measured at about 25° scattering angle, surprisingly increased unfilterable haze measured at about 90° scattering angle, as the mean particle size was reduced by this treatment

(Van Roon et al . , 31° EBC Congress, 2007).

[0017] Therefore, "unfilterable haze" remains a major problem in today's brewing practices. Aims of the invention

[0018] A first aim of the invention is to provide a new method and composition that does not present the drawbacks of the methods and compositions of the state of the art.

[0019] A preferred aim of the invention is to propose such method and composition that could reduces significantly "unfilterable haze" in fermented beverages, especially beer, wine and cider, particularly so called "pils" type beer or white wine, especially an unfilterable haze measured at about 90° scattering angle.

Summary of the invention

[0020] The present invention relates to a method to reduce haze in a (fermented) beverage by adding to this beverage, a sufficient amount of an enzyme (composition) , this enzyme composition comprising or consisting

(essentially) of an alpha-glucanase able to work from about

0⁰C to about 10⁰C and deactivated by the beverage pasteurization.

[0021] The preferred (fermented) beverages of the invention are beer, wine and cider, more preferably so called "pils" bier or white wine. [0022] Preferably, the method and composition of the invention allows a reduction of "unfilterable haze" in this beverage (measured using EBC standards and different light scattering angles (about 25° and about 90°). [0023] Preferably, the "unfilterable haze" does not develop a blue coloration after reaction with iodine. [0024] Advantageously, the method and enzyme (composition) of the invention allows a reduction of "unfilterable haze" to values compatible with brewer' s norms, being correlated with values from about 0.5 to about 0.7 EBC units measured with a side scattering angle of

(about) 90°.

[0025] Alternatively, the method and enzyme

(composition) of the invention allows an at least two-fold reduction of "unfilterable haze", preferably as measured using EBC standards with a side scattering angle of (about)

90°.

[0026] The method of the invention may also comprise a "chill proofing" step with the addition of a sufficient amount of fining agents, preferably selected from the group consisting of PVPP, silicagel, tannins, proteases (not aspartyl and not proline-specific) or a mixture thereof.

[0027] By Consisting essentially' , it is meant that alphaglucanase activity has to be present. The lowest recommended activity is 1000 DU/g.

[0028] By alphaglucanase, it is meant an enzyme

(composition) able to hydrolyse alpha-glucans, that are OCi-4 and OCi-6 glucose polymers, being preferably an alpha amylase or an amyloglucosidase, more preferably an alpha amylase.

[0029] Alternatively, another preferred alphaglucanase is a pullulanase and more preferably a pullulan-6-glucanohydrolase (EC .3.2.1.41) or alpha 1,6 pullulase . [0030] One dextrinizing unit (DU) is defined as the quantity of α-amylase (alphaglucanase) that will dextrinize soluble starch in the presence of an excess of β-amylase at the rate of 1 g/h at 30⁰C.

[0031] Preferably, the enzyme (composition) according to the invention has mainly an alphaglucanase activity of about 1000 DU/g to about 100000 DU/g, more preferably of about 30000 DU/g. [0032] Advantageously, the alphaglucanase of the invention used during beer maturation or in BBT (bright beer tank or tank of filtered beer) does not modify organoleptic properties of this beverage (alcohol content, flavour, taste, foam stability, ...)•

[0033] The other (s) element (s) possibly present in the composition is (are) (an) other enzyme (s) or one or more usual carrier (s) of enzyme (s).

[0034] Advantageously, the (enzyme) composition of the invention further comprise a sufficient amount of a beta-glucanase (cellulase; EC. 3.2.1.4), preferably having an activity comprised between about 1 and about 1000 BGU/g, more preferably comprised between about 10 and about 500 BGU/g, still more preferably of about 200 BGU/g.

[0035] One β-glucanase unit (BGU) is defined as the quantity of enzyme that will liberate reducing sugar (as glucose equivalent) at a rate of 1 mmol/min at 40⁰C and pH 6.5.

[0036] Preferably, the (enzyme) composition of the invention further comprises a sufficient amount of a xylanase (endo 1,4 beta xylanase EC.3.2.1.8), more preferably having an activity between about 1 and about 1000 FXU/g more preferably comprised between about 10 and about 500 FXU/g, still more preferably of about 100 FXU/g. [0037] Endoxylanase activity is measured relative to an enzyme (xylanase) standard and calculated in Farvet Xylanase Units (FXU) .

[0038] More preferably, the (enzyme) composition of the invention further comprises a sufficient amount of a protease, more preferably having an activity comprised between about 1 and about 200000 HUT/g, more preferably comprised between about 10 and about 20000 HUT/g, still more preferably comprise between about 100 and about 5000 FXU/g.

[0039] One HUT unit of proteolytic (protease) activity is defined as that amount of enzyme that produces, in 1 min at 40⁰C and pH of 4.7, a hydrolysate whose absorbance at 275 nm is the same as that of a solution containing 1.10 μg per ml of tyrosine in 0.006 N hydrochloric acid. [0040] The protease of the invention is preferably an aspartic protease.

[0041] Alternatively, the protease of the invention is a proline-specific protease.

[0042] Advantageously, the proteases of the invention consists of both an aspartic protease and a proline-specific protease.

[0043] Preferably, all the enzymes (alphaglucanase, beta-glucanase, xylanase and protease) used in the present invention are from fungal origin, more preferably are obtained from Aspergillus . [0044] Advantageously, the alphaglucanase can be (further) obtained from a prokaryotic origin. [0045] Another preferred composition of the invention comprises (or consist of) a sufficient amount of the alphaglucanase and a sufficient amount of a protease, possibly with a sufficient amount of a beta glucanase and possibly with a sufficient amount of a xylanase. [0046] A last preferred composition of the invention comprises (or consist of) a sufficient amount of the alphaglucanase of the invention and a sufficient amount of a xylanase.

[0047] The preferred beverages according to the present invention are beverages involving a fermentation step using yeasts, such as beer, wine and cider, the most preferred beverages being (pils) beers. [0048] Preferably, the enzymes of the invention (alphaglucanase, and possibly beta-glucanase, protease (being preferably aspartyl protease) and/or xylanase) are added during the maturation step of this beverage, being preferably beer, wine or cider.

[0049] The preferred enzymes (alphaglucanase and possibly, beta-glucanase, protease (being preferably aspartyl protease) and/or xylanase) are active at temperatures comprised between about 0⁰C and about 10⁰C and at an acidic pH comprised between about 4.0 and about 6.0, preferably at an acidic pH of about 4.2.

[0050] Advantageously, the method of the invention comprises (or consists of) a fermentation step of a beverage, with or without the addition of a proline-specific protease a maturation step of this beverage, wherein this beverage is cooled at a temperature comprised between about 0⁰C and about 10⁰C, possibly a "chill proofing" step and treatment with addition to the beverage of fining agents (such as PVPP, silicagel, tannins) and/or proteases to minimize chill haze possibly a standing in BBT (bright beer tank) abtween about 0 and about 5°C waiting for the bottling or kegging step (from a few hours to 60 hours) possibly a pasteurization step of this beverage and possibly a collecting step of the obtained beverage in a suitable container (bottle, barrel, etc) , wherein a sufficient amount of the enzyme

(composition) of the invention is added to the beverage during this fermentation and/or this keeping (maturation and BBT) step(s) for a preferred delay comprised between about 1 hour and about 10 days. [0051] Preferably, in the method of the invention, the sufficient amount of the enzyme (composition) is comprised between about 0.10 g and about 10 g per degree plato (⁰P) (original wort gravity) and per hectolitre. [0052] Preferably, in the method of the invention, the enzyme (composition) is added to this beverage during the maturation step of this beverage, preferably at a temperature comprised between about 0⁰C and about 10⁰C.

[0053] Advantageously, the enzyme (composition) of the invention (alphaglucanase and possibly, xylanase, beta- glucanase and/or protease (being preferably aspartyl protease) ) is denaturated during a pasteurization step, and its residual activity is reduced by at least 90%, preferably by at least 95%, more preferably by at least 99%.

[0054] Advantageously, the enzyme (composition) of the invention (alphaglucanase and possibly, xylanase, beta- glucanase and/or protease) used during beer maturation or in BBT (bright beer tank or tank of filtered beer) , does not modify the organoleptic properties of this beverage (alcohol content, flavour, taste, foam stability, ...) [0055] Alternatively, the enzyme (composition) of the invention (alphaglucanase, and possibly beta-glucanase, protease (being preferably aspartyl protease and/or a proline-specific protease) and/or xylanase) are added to this beverage during fermentation step of the beverage

(preferably beer, wine and cider) . [0056] Alternatively, the enzyme (composition) of the invention (alphaglucanase, and possibly beta-glucanase, protease (being preferably aspartyl protease) and/or xylanase) is added to this beverage during bright beer keeping step (BBT tank) just before bottling or kegging beer. The method of the invention may also comprises the additional step of adding to the beverage of one or more fining agent (s), preferably selected from the group consisting of PVPP, silicagel, tannin, protease or a mixture thereof.

Brief description of the figure

[0057] Figure 1: Evolution of beer haze during fermentation without and with the addition of the enzyme composition of the invention.

[0058] Measurements of haze produced during fermentation are performed after filtration of a sample of fermenting beer on kieselguhr at laboratory scale and by measuring haze with a side scattering angle of 90°.

Detailed description of the invention

[0059] The preferred enzyme composition of the invention comprises (or consists of) about 30000 DU/g of alphaglucanase, about 200 BGU/g of beta-glucanase, about 100 FXU of xylanase and about 5000 HUT/g of protease.

[0060] Advantageously, the assay to determine the α- amylase activity of enzyme preparation is based on the time required to obtain a standard degree of hydrolysis of a starch solution at 30⁰C ± 0.1⁰C. The degree of hydrolysis is determined by comparing the iodine colour of the hydrolysate with that of a standard.

[0061] Advantageously, the assay used to determine β-glucanase activity of enzyme preparation is based on a 15-min hydrolysis of lichenin substrate at 40⁰C and at pH 6.5. The increase in reducing power due to liberated reducing groups is measured by the neocuproine method. The absorbance of each solution is determined at 450 nm. [0062] Advantageously, the assay used to determine endoxylanase samples is based on the use of remazol-stained wheat arabinoxylan substrate. Unconverted substrate is precipitated with ethanol. The intensity of the blue colouring of the supernatant due to unprecipitated remazol- stained substrate degradation products is proportional to the endoxylanase activity. Xylanase activity is measured relative to an enzyme standard and calculated in Farvet Xylanase Units (FXU) . A stock solution is used to prepare several FXU standard working solutions and to give a range of activities for the construction of the standard curve. [0063] Advantageously, the assay used to determine proteolytic activity in a sample is based on a 30-min enzymatic hydrolysis of a haemoglobin substrate at pH 4.7 and 40⁰C. Unhydrolyzed substrate is precipitated with trichloroacetic acid and removed by filtration. The quantity of released tyrosin is determined spectrophotometrically . A Standard Curve is drawn using chromatographic-grade (ultra-pure) L-tyrosine. [0064] This (preferred) enzyme composition of the invention is added at about 0.10 to about 1 g per ⁰P (degree plato) (original wort gravity) and per hectolitre.

[0065] By degree plato (⁰P), it is meant the weight percentage of dry matter (mainly sugar) in this beverage

(preferably beer, wine or cider) . By original wort gravity it is meant the gravity of the wort at the beginning of fermentation expressed in degree Plato (⁰P) .

[0066] The skilled one knows how to convert degree Plato (⁰P) into degree Brix (⁰B), as the dry matter consists essentially on sugar, these two measures are almost equivalent. In the present invention, it is assumed that about 1°P equates about 0.95°B.

[0067] The skilled one is also aware of the Balling scale and is able to convert values in ⁰P into Ballings scale, these two values being almost equivalent. [0068] Advantageously, the enzyme (composition) and/or the method of the invention allows an efficient reduction of beverage "unfiltrable haze" at levels lower to

(about) 0.75 EBC (side scattering angle of about 90°), preferably lower than (about) 0.70 EBC (side scattering angle of about 90°), more preferably lower than about 0.60

EBC (side scattering angle of about 90°) .

[0069] By EBC units (as units accepted by the European brewery convention, preferably using formazin suspensions as standards) , it is meant a turbidity measure with a forward scattering angle of about 25° (or about 13° according to EBC standard) for evidencing common (chill) haze or side-scatter angle of about 90° for evidencing lower values of haze. Brewing organizations such as the European Brewing Convention (EBC) and the American Society of Brewing Chemists (ASBC-FTU) employ their own formazin based turbidity unit scales. While EBC units are commonly used for both forward and side-scatter measurements, ASBC FTU is strictly a 90° measurement. [0070] This validated (EBC) measure is colour- independent .

[0071] Other measures are developed and the skilled person knows how to convert these units obtained following EBC standards to units obtained following other standards. For instance, 1 EBC (European Brewery Convention) is equivalent to 4 TEF (Trubungseinheiten Formazin) , and equivalent to 40 Helm units, equivalent to 69 ASBC (American Society of Brewing Chemists) [0072] The EBC measure for assessing the "unfiltrable haze" is done using EBC requirements, but the turbidity is measured with a side scattering angle of about 90°. Examples

Example 1 : validation of "unfilterable haze" measures and effect on a beer quality [0073] "Unfilterable haze" was measured using EBC standards with a side scattering angle of 90°. The inventor tested several methods and conclude that the use of EBC standards with a side scattering angle at 90° better evidences "unfilterable haze". [0074] Experimented brewers judged several beers having no (common) (chill) haze (PVPP, silicagel, papain, or Clarex treated) , but only "unfilterable haze" with known EBC values measured using a side scattering angle of 90°. For these beers, these panellists concluded that values above about 0.75 EBC (side scattering angle of 90°) presented a risk of colloidal instability during beer staling.

Example 2 : reduction of "unfilterable haze" by the enzyme composition of the invention in 3 different beers chill- proofed and treated with PVPP

[0075] Three different pils beers were produced using mashes of different sugar content (see Table 1, left col., ⁰P being mostly function of the sugar content) and with a fermentation step carried out at different temperatures (see Table 1, second col.).

[0076] These three beers were chilled at below 0⁰C, treated with PVPP (Divergan BASF, 30 g/hl) and filtered in order to remove the common (chill) haze. [0077] As shown in the third col. of Table 1, all the values measured with a side scattering angle of 90° are well above the limit of about 0.75 EBC. and are therefore judged as of poor quality. [0078] The inventor then applied to the beers 250 mg/°P beer original gravity/Hi of the (preferred) enzyme composition of the invention (comprising about 30000 DU/g of alphaglucanase, about 200 BGU/g of beta-glucanase, about 100 FXU of xylanase and about 5000 HUT/g of protease), during the maturation step of these beers at about 4⁰C.

Table 1: reduction of "unfilterable haze" by the enzyme (composition) of the invention in 3 different beers treated with PVPP at the rate of 30g/hl; turbidity values are in EBC and measured with a scattering angle of 90°

[0079] As shown in Table 1, the inventor measured a strong reduction in "unfilterable haze" and conclude that all the three treated beers now correspond to the standards for commercialization.

[0080] The inventor further tested a composition consisting essentially of an alphaglucanase (from prokaryotic origin being from Bacillus acidopullulyticus) with a specific activity of 1000 DU/g and substantially free of beta-glucanase, xylanase and protease side- activities. When using this simplified composition, the inventor obtained a consistent reduction in unfliterable haze, albeit somehow less spectacular than when using the composition from fungal origin (further comprising about beta-glucanase, xylanase and about protease) . Example 3 : reduction of "unfilterable haze" by the (preferred) enzyme composition of the invention in 3 different beers chill-proofed and treated with silcagel [0081] The same three beers as in example 2 were chilled at below 0⁰C, treated with silicagel (Spindal Polygel from Spindal AEB Group, 40 g/hl) and filtered in order to remove the common (chill) haze.

[0082] As shown in the third col. of Table 2, all the values measured with a side scattering angle of 90° are well above the limit of about 0.75 EBC and are therefore judged as of poor quality.

[0083] The inventor then applied to the beers 250 mg/°P beer original gravity/Hi of the (preferred) enzyme composition of the invention, during the maturation step of these beers at about 4°C.

Table 2: reduction of "unfilterable haze" by the enzyme (composition) of the invention in 3 different beers treated with silicagel at the rate of 40g/hl; turbidity values are in EBC and measured with a scattering angle of 90°.

[0084] As shown in Table 2, the inventor measured a strong reduction in "unfilterable haze" and conclude that all the three treated beers now correspond to the standards for commercialization. Example 4 : reduction of "unfilterable haze" by the (preferred) enzyme composition of the invention in 3 different beers chill-proofed and treated with papain [0085] The same three beers as in example 2 were chilled at below 0⁰C, treated with papain (Papain-Clarase from CBS Chemicals, 4g/hl) and filtered in order to remove the common (chill) haze.

[0086] As shown in the third col. of Table 3, all the values measured with a side scattering angle of 90° are well above the limit of about 0.75 EBC and are therefore judged as of poor quality.

[0087] The inventor then applied to the beers 250 mg/°P beer original gravity/Hi of the (preferred) enzyme composition of the invention, during the maturation step of these beers at about 4°C

Table 3: reduction of "unfilterable haze" by the enzyme (composition) of the invention in 3 different beers treated with papain 100 TU at the rate of 4g/hl; turbidity values are in EBC and measured with a scattering angle of 90°

[0088] As shown in Table 3, the inventor measured a strong reduction in "unfilterable haze" and conclude that all the three treated beers now correspond to the standards for commercialization. Example 5 : reduction of "unfilterable haze" by the enzyme composition of the invention in 3 different beers chill- proofed and treated with a proline-specific protease PSP [0089] The same three beers as in example 2 were treated with a PSP proline-specific protease at pitching (Clarex from DSM, at the concentration of 20 PPU/hl) and were chilled at below 0⁰C, and filtered in order to remove the common (chill) haze.

[0090] As shown in the third col. of Table 4, all the values measured with a side scattering angle of 90° are well above the limit of about 0.75 EBC and are therefore judged as of poor quality.

[0091] The inventor then applied to the beers 250 mg/°P beer original gravity/Hi of the (preferred) enzyme composition of the invention, during the maturation step of these beers at about 4°C

Table 4: reduction of "unfilterable haze" by the enzyme (composition) of the invention in 3 different beers treated with a proline-specific protease PSP at the concentration of 20 PPU/hl; turbidity values are in EBC and measured with a scattering angle of 90°.

[0092] As shown in Table 3, the inventor measured a strong reduction in "unfilterable haze" and conclude that all the three treated beers now correspond to the standards for commercialization.

Example 6 : the enzyme composition of the invention has been added in cylindro-conical fermenters (CCT) pitched with freshly rehydrated dry yeast

[0093] The use of freshly rehydrated dry yeast always leads to a abnormal higher haze in the beer. Therefore the inventor has investigated the effect of addition of the (preferred) enzyme composition of the invention at the beginning of fermentation on the haze produced during fermentation with freshly rehydrated yeast. [0094] Fermentations were carried out at 16°C and 1560 Hl of wort at 14.5°P were pitched with 150 kg of freshly rehydrated yeast. In total 5.6 kg of the enzyme composition of the invention were added in each CCT2. A control CCTl without addition of enzyme was also followed after pitching with 150 kg of freshly rehydrated yeast. [0095] The results for both CCTs are illustrated in Figure 1 and clearly indicate the positive effect of the enzyme composition of the invention on the haze production during fermentation.

Example 7 : effect of the enzyme composition of the invention on white wine

[0096] The addition of the (preferred) enzyme composition of the invention to a white wine fermented at 16⁰C (11% of alcohol by volume) treated with PVPP allowed to decrease the haze measured with a side scattering angle of 90° from 1.8 EBC to 0.3 EBC.

Claims

1. A method to reduce haze in a beverage by adding a sufficient amount of an enzyme

(composition) comprising an alphaglucanase to the said beverage.

2. The method of claim 1, wherein the enzyme (composition) consists (essentially) of an alphaglucanase .

3. The method of claims 1 or 2, wherein the haze is unfilterable haze.

4. The method of claim 3, wherein the unfilterable haze is measured using EBC standards and different light scattering angles of 25° and/or of 90°.

5. The method of claims 3 or 4, wherein the unfilterable haze is measured using EBC standards and light scattering angles being a side scattering angle of (about) 90°.

6. The method according to any of the preceding claims 3 to 5, wherein the unfilterable haze does not develop a blue coloration after reaction with iodine .

7. The method according to any of the preceding claims 1 to 6, wherein the beverage is selected from the group consisting of beer, wine or cider .

8. The method according to any of the preceding claims 1 to 7, wherein a sufficient amount of a beta-glucanase enzyme is further present in the enzyme composition.

9. The method according to any of the preceding claims 1 to 8, wherein a sufficient amount of a protease enzyme, preferably an aspartyl protease enzyme, is further present in the enzyme composition.

10. The method according to any of the preceding claims 1 to 9, wherein a sufficient amount of a proline-specific protease enzyme is further present in the enzyme composition.

11. The method according to any of the preceding claims 1 to 10, wherein a sufficient amount of a xylanase enzyme is further present in the enzyme composition.

12. The method according to any of the preceding claims 1 to 11, wherein the enzyme composition is of fungal origin.

13. The method according to the claim 11, wherein the enzyme is from Aspergillus .

14. The method according to any of the preceding claims 1 to 11, wherein the alphaglucanase is a pullulanase.

15. The method of claim 14, wherein the pullulanase is a pullulan-6-glucanohydrolase (EC.3.2.1.41) .

16. The method according to any of the preceding claims 1 to 15, wherein the added amount of the enzyme (composition) is comprised between 0.10 g and 10 g per degree plato (⁰P) and per hectolitre of beverage .

17. The method according to claim 16 wherein the enzyme (composition) encompasses an alphaglucanase activity of about 1000 DU/g to about 100000 DU/g.

18. The method according to any of the preceding claims 1 to 17, wherein the sufficient amount of the enzyme (composition) is added during beverage maturation step, preferably at a temperature comprised between 0⁰C and 10⁰C.

19. The method according to Claims 17 or 18, wherein the beverage is beer and the sufficient amount of enzyme (composition) is added during the bright beer keeping (BBT tank) just before bottling or kegging at a temperature comprised between 0⁰C and 10⁰C.

20. The method according to any of the preceding claims, wherein the (unfilterable) haze is reduced by at least two fold.

21. The method according to any of the preceding claims, wherein the organoleptic properties of the beverage are not modified.

22. The method according to any of the preceding claims 1 to 21, which further comprises a step of adding a sufficient amount of one or more fining agent (s) selected from the group consisting of PVPP, silicagel, tannins, proteases (not aspartyl and not proline-specific) or a mixture thereof to the beverage .