ENZYMES IN THE BREWERY

Why and when to use exogenous enzymes in the brewery?

Most of the enzymes which are available on the market are natural products as malt enzymes are. Commercial enzymes are mainly produced by fermentation.

The enzymes are quite simply proteins which have, in addition to their catalytic activity, structures and biochemical properties similar to other commonly known proteins. They are used in many food processes. The quantities used in the production of foodstuffs are always very low and their catalytic activities are inactivated during different successive stages of manufacture. For these reasons, the European Legislation classifies enzymes as processing aids and not as food additives.

The use of exogenous enzymes in brewery must be considered on the basis of some quite precise criteria:

– increase of raw materials extract yields
– time saving and thus greater productivity
– maintenance or improvement of the quality of produced beer
– adaptation of the manufacturing process:

– to produce a new product
– to the use of new raw materials.

The productivity and raw materials extract yield optimization implies also the necessary adaptation to the seasonal variations of raw materials quality.

The increase of productivity and of raw materials extract yield has to allow to save money including enzyme cost.

The improvement of one of the three criteria: productivity, yield and quality could already justify the use of an exogenous enzymatic preparation.

IN THE BREWHOUSE

How to increase raw materials extract yield, wort and beer
filtration rate and beer colloidal stability?

Raw materials like barley, malt, wheat, oats, rye contain in addition to starch several different polysaccharides which contribute to wort and beer viscosity and to beer colloidal stability.The most important polysaccharides besides starch in barley and malt are ß-glucans which constitute more than 75% of cell walls. Malt contains three times more soluble pentosans than barley. Pentosans are wheat main polysaccharides in addition to starch and their content is more than two times higher than in malt. Insoluble pentosans are important constituents of spent grains. They have the ability to absorb water quantities equivalent to ten times their own weight. They contribute mainly to spent grains spongy and sticky character.

Exogenous ß-glucanases and pentosanases used during mashing allow to lower wort and beer viscosity and therefore to increase wort and beer filtration rate. Spent grains manipulation also becomes easier. Furthermore, higher ß-glucans and pentosans hydrolysis contributes to improve starch extraction and raw material extract yield as these polysaccharides are the main constituents of endosperm cell walls.

Proteins can also originate filtration problems by their ability and trend to form gels. Protease addition during brewing can improve wort filtration rate by avoiding proteinic gels formation. On the other hand a protease can also solubilize ß-glucans bonded to cell walls proteins by covalent linkages. In this case a ß-glucanase has to be used to hydrolyze ß-glucans solubilized by this protease. Extract yield is also improved by the use of proteases.

Glucanylase is a formulation studied to meet the three following criteria: improving of extraction yield, increase of productivity and higher beer quality. Glucanylase contains ß-glucanases, pentosanases, proteases and thermostable alpha-glucanases. ß-glucanases, pentosanases and proteases improve solubilisation of cellular walls of endosperm cells allowing a better starch extraction. These three enzymes increase brewhouse extract yield and agree with first criterion. ß-glucanases and pentosanases act on ß-glucans and gums and improve wort and beer filtration rate (increase of productivity: second criterion). Proteases prevent proteinic gels formation which can involve problems during wort filtration. Last but not least ß-glucanases, pentosanases and thermostable alpha-glucanases contribute to a better beer colloidal stability (third criterion). In fact the presence of higher ß-glucans, gums and/or higher alpha-glucanes in beer can be a source of colloidal disorder.

Small starch granules gelatinise at higher temperatures than normal size granules of a given cereal. It can occur that some small granules are not gelatinised at the end of brewing and are retained in spent grains during filtration or reach the following stage and gelatinise during wort boiling.

In the first case, iodine coloration test applied to spent grains will give a blue coloration translating the presence of higher alpha-glucans and therefore a loss of hydrocarbonated extract yield.

In the second case, small granules passing the stage of wort filtration will gelatinise during wort boiling whereas malt alpha-amylases are already destroyed. Higher alpha-glucans (iodine coloration test on wort after boiling is blue) will remain up to final beer involving risks of colloidal instability. The presence of thermostable alpha-glucanase when using GLUCANYLASE allows to raise mash temperature as final brewing step to gelatinise smaller starch granules while liquefying them by this thermostable enzyme.

The iodine coloration test of spent grains should then be negative and the consequence is an increase of the hydrocarbonated extract (first criterion). If in spite of everything higher alpha-glucans were to pass the filtration stage, they would be liquified by GLUCANYLASE thermostable alpha-glucanase which remains still active a few minutes in boiling tank. The presence of higher alpha-glucanes in beer is avoided just as the problems of colloidal type which would be dependent on them (third criterion).

Beer filtration problems have to be solved during brewing. Often beer filtration difficulties were preceded by a bad wort filtration. Sometimes the use of mash tuns making strongly turn the knives, or the most powerful techniques of wort filtration such as mash filter, 2001 filter, Nordon filter etc. can mask the tendency of a wort to filter badly and the problems are then deferred on beer filtration. The filtrability of beer depends inter alia on the presence of polysaccharides such as alpha-glucans, ß-glucans and pentosans. Some of these polysaccharides can pass the stage of final filtration and can be the cause of colloidal disorder. The adequate processing of alpha-glucans, ß-glucans and pentosans during brewing improves beer filterability and contributes to a good beer colloidal stability.

WORT FILTRATION
—> ß-glucans
—> pentosans
—> alpha-glucans
—> proteins
—> ß-glucanases
—> pentosanases
—> themostable alpha -glucanases
—> proteases
During brewing
During brewing
During brewing
During brewing
BEER FILTRATION
—> ß-glucans
—> pentosans
—> alpha-glucans
—> proteins
—> ß-glucanases
—> pentosanases
—> themostable alpha -glucanases
—> proteases
During brewing
During brewing
During brewing
During brewing
COLLOIDAL STABILITY
—> ß-glucans
—> pentosans
—> alpha-glucans
—> proteins
—> polyphenols
—> ß-glucanases
—> pentosanases
—> themostable alpha -glucanases
—> papain, silicagel, tannins, etc.
—> PVPP
During brewing
During brewing
During brewing

 

BEER BRAND FINAL TOUCH

CBS Customized Brewing Solutions supplies a product line named « Brand Final Touch » allowing to produce « special beers » by adding to a normal pale beer just before bottling.

With our Brand Final Touch range of products, we can produce the following beers from a unique neutral lager: Premium Beer, Witbier, Abbey Beer, Sour Ale, Weizenbier, Stout, IPA, Grapefruit Beer, Cherry Beer, Raspberry Beer, Strawberry Beer, Mojito Beer, Sweet Rhum Beer, Daiquiri Beer, Lychee Beer, Peach Beer, etc. For more information, contact our technical team.

All our « final touch » beer brand producers contain exclusively natural aroma compounds (fruits and spices aroma compounds), natural colouring (fruit colours and dark malt extracts) and clouding agents (essential oils emulsions stabilised with vegetal gums).

WB-MIX has been set up to produce white beers with taste, flavour and cloudy aspect similar to main commercial Belgian white beers which are refreshing and thirst-quenching. WB-MIX has to be added to a standard base beer with low bitterness (10-12 EBU) as unique requisite to obtain a white beer. This brand maker can be customized on demand. (WB-MIX Project).

PF-MIX-HOP range of products allows to improve the palate fullness, the freshness and the hop aroma of the base beer. It improves the drinkability and delays the appearence of staling off-flavours. The resulting beer is a well balanced Premium Beer with increased shelf life..

Among other CBS brand producers the following could be mentioned: KB-MIX (fruits and spices colour and aroma compounds) to produce cherry beer (kriek-like), fruits and spices colour and aroma compounds for raspberry beer, fruits and spices aroma compounds for peach beer, Spring-MIX (spices aroma compounds), PF-MIX to impart palate fullness and a long list of customized brand makers (brown beer producer, ginger ale producer, etc).

All these brand producers can be used to produce low alcohol content (as low as 0.5% v/v) special beers (low alcohol white beer, kriek, etc).

BASE BEER : Low colour, 7-8% alcohol, 12-14 EBU

1) WATER RECTIFICATION

2) BITTERNESS ADJUSTEMENT

Substitution of part of classical hopping by a product isomerized at high concentration can contribute to improve beer staling resistance. (Anti-Staling).

3) COLOUR ADJUSTEMENT

Beer colouring with dark malt extract is in high demand by dose marketing their beer on an « additive free » platform.

4) FLAVOUR ADJUSTEMENT

HOP FLAVOUR : IsototalHopessoil-HALPF-MIX-HOPPF-MIX-HOP-NAPF-MIX-HOP12PF-MIX-HOP12-NA
WHITE BEER FLAVOUR : WB-MIXWB-MIX project
KRIEK (CHERRY) BEER FLAVOUR : KB-MIX
LYCHEE FLAVOUR : LYCHEE-MIXLYCHEE-MIX-C
PALATE FULLNESS : PF-MIXMalolactic MIX
RASPBERRY FLAVOUR : RASP-MIX-S
OTHERS : AB-MIX – Premium Beer – Witbier – Abbey Beer – Sour Ale – Weizenbier – Stout – IPA – Grapefruit Beer – Cherry Beer – Raspberry Beer – Strawberry Beer – Mojito Beer – Swee Rhum Beer – Daiquiri Beer – Lychee Beer – Peach Beer – etc.
All our flavours mixes can be customized. They contain exclusively natural compounds (fruits, spices, malt extract).

ATTENUATION MODULATION

How to adjust wort fermentability?

A wort produced by conventional brewing will show an apparent attenuation around 80-85% and almost 25% of carbohydrate extract will be present in final beer as non fermentable short chain dextrins.Sometimes, the brewer would like to increase this wort fermentability to obtain an apparent attenuation of 85-90%, 90-95%, 95-100%, = 100% or even > 100%. For a given alcohol level, the more attenuated beer the lower original gravity and therefore the lower raw material quantity involved. The traditional brewing methods are not very efficient to reach these aims. The use of appropiate exogenous enzymes allows to modulate almost as much as desired wort sugars profile and therefore wort fermentability as far as carbon hydrates are concerned.Different exogenous enzymes are available to modulate wort fermentability : amyloglucosidase (Glucosylase), pullulanase or limit dextrinase (Desatase), and maltogenic amylases alpha (Maltosylase) or beta.Amyloglucosidase (Glucosylase) is an exo-amylase which catalyses starch dextrins, oligosaccharides and long chains hydrolysis from non reducing ends to produce glucose units by hydrolysing quickly alpha-1,4 linkages and slowly alpha-1,6 linkages.
Pullulanase or limit dextrinase (Desatase), catalyses quick hydrolysis of alpha-1,6 linkages of starch dextrins, oligosaccharides and long chains.
Maltogenic alpha-amylase (Maltosylase) is an endo-amylase that hydrolyses at random alpha-1,4 bonds located in the inner regions of the starch substrate removing maltose units at each hydrolysis on a multi-chain action.
Maltogenic beta-amylase is an exo-amylase which hydrolysis starch chains from the non-reducing ends producing successive maltose units on an uni-chain action.
These enzymes may be used either in the brewhouse or during fermentation depending on the results expected. When used during brewing these four different enzymes will be inactivated during wort boiling.
When used during fermentation, residual activity will vary from an enzyme to another as well as enzyme shelf-life at 60°C.

Desatase Maltosylase Glucosylase

Residual enzymatic activity in beer after 7 days of fermentation,
after 30 days of maturation and shelf-life in beer at 60°C

Maltogenic α-amylase 20% residual enzymatic activity after 7 days of fermentation
0% residual enzymatic activity after 30 days of maturation
1 min shelf-life at 60°C in beer
Maltogenic ß-amylase 20% residual enzymatic activity after 7 days of fermentation
0% residual enzymatic activity after 30 days of maturation
1 min shelf-life at 60°C in beer
Pullulanase 40% residual enzymatic activity after 7 days of fermentation
0% residual enzymatic activity after 30 days of maturation
1 min shelf-life at 60°C in beer
Amyloglucosidase 100% residual enzymatic activity after 7 days of fermentation
100% residual enzymatic activity after 30 days of maturation
60 min shelf-life at 60°C in beer


Equivalence of the enzymes here above and CBS trade names:

Maltogenic alpha-amylase : Maltosylase
Pullulanase : Desatase
Amyloglucosidase : Glucosylase
Maltogenic ß-amylase + Pullulanase MIX : Maxiferm

 

This Table explains how to adjust wort fermentability by choosing judiciously one or more enzymes and where to apply them.

Apparent
Attenuation
Enzyme
Name
Dose
Point of addition
Residual activity
in beer
85-90%
Maltosylase-alpha
0.5 to 5 g/hl
Start of fermentation
no active enzyme
in pasteurized beer
90-95%
Glucosylase+
Desatase
1-3 l/ton of grist
3-5 kg/ton of grist
At mashing in
At mashing in
no active enzyme
during fermentation
95-100%
Desatase+
Maltosylase
1-3 kg/ton of grist
4-8 g/hl of beer
At mashing in
Start of fermentation
no active enzyme
in pasteurized beer
=100%
Desatase+
Maltosylase
1-3 kg/ton of grist
2-5 g/hl of beer
4-8 g/hl of beer
At mashing in
Start of fermentation
no active enzyme
in pasteurized beer
>100%
Glucosylase
3-5 g/hl of beer
Start of fermentation
no active enzyme
in pasteurized beer

 

Notes : 1. Alpha-amino nitrogen is supposed to be in sufficient concentration and is not restrictive
2. The use of Glucosylase generates glucose. This last may cause loss of ability to uptake maltose by some sensitive yeast strains

ANTI-STALING

Improvement of resistance against beer staling

Last scientific researches confirm that the staling-potential of a beer is essentially determined in the brewhouse. Therefore it is very important to act as soon as possible in the brewing process.

First of all by trying to minimize oxygen presence (flours, brewing water, brewing under CO2, etc) and by decreasing malt lipoxygenase action during brewing (unsaturated lipids oxydation to lipid peroxides that can oxidize proteins and that can act during wort boiling as precursors of compounds like trans-2-nonenal).
After that care has to be taken during wort boiling to minimize oxidation processes and especially lipids oxidation (peroxides lipids). Oxidation reaction products like trans-2-nonenal (cardboard taste) can in the first place establish bounds with compounds like sulfites for example that make them imperceptible in just bottled beer. Later, these sulfites become gradually oxidized in the beer bottle releasing the undesirable compounds that become perceptible at this moment.

The use of an antioxidant and natural reducing agent at mashing-in (Tannox-Brew) allows to decrease malt lipoxygenase action (more than 50%). In fact lipoxygenase needs its core to be at ferric state (Fe3+) as well as molecular oxygen presence to oxidize lipids into peroxides. Ferric core reduction into ferrous iron deactivates this enzyme. The decrease of oxygen disponibility slackens lipoxygenase action. TANNOX-BREW antioxidant action also decreases proteins oxidation and therefore increases wort filtration rate. The use of classical antioxidants like ascorbic acid-sulfites couple during wort boiling completes this first anti-staling action in the brewhouse.

Tannox-Brew

During wort boiling hop is added to perform isomerisation step. Hop also can then be oxidized giving rise to several compounds that can significantly participate to typical tastes and flavours faults of stale beers. Separated isomerisation of a very concentrated hop extract solution (a minimum of 20% of alpha acids in solution) allows to minimize hop oxidation thanks to natural hop reducing power. This reducing power is significant when isomerisation is performed on a concentrated hop extract solution but it is not sufficient at usual concentration in wort. Substitution of a part of classical hopping by a product like Isototal isomerized at high concentration can also contribute significantly to improve beer anti-staling resitance.

Fight against oxygen presence in beer has to be continued by classical ways during bottling and by adding an antioxidant to finished beer.

Isototal

WB-MIX PROJECT

WB-MIX project began in 1997. The aim was to provide a method to produce a Belgian type white beer by privileging production flexibility allowing a quick response to new markets demands.

WB-MIX is the result of a market survey, a marketing strategy, fundamental search on flavours (beers and spices), adjustment of search results to the consumer (tastings) and the follow-up of beers produced by this way since 1997.

WB-MIX-1 formula setting was carried out after making a survey of the reasons of the success of white beer in Belgium that passed from being a village beer in 1978 to be currently produced at the rate of 1 000 000 Hl/year. This success is due to the fact that a whole rising generation liked this low bitter beer flavoured with spices. It was an alternative to classical beer taste, between beer and soft drink. Moreover, the product was well differentiated by its turbid aspect giving rise to a fashion trend similar to that which occurred with a popular Mexican beer in colourless bottle and served with a lemon piece in the bottle neck. The Belgian type white beer pleased the people who did not like so much normal beer and who wanted to drink another thing than soft drinks.

Our target was thus a person who did not like traditional beer and who was not a white beer expert. We analyzed 60 white beers (see Table below). On these values basis and the results of tasting pannels established according to our target definition, we composed a series of WB-MIX. Then we made consumer tests (hypermarkets) in countries with low culture of white beers. WB-MIX-1 came out due to its high « drinkability ». This formula can however be customized according to the own criteria of the brewer.

The use of WB-MIX offers several advantages in comparison with the traditional method which involves the use of spices in the brewhouse and « special » yeast strains producing the metabolites which impart to beer typical white type flavour and taste (essentially phenolic compounds: vinylguaïacol, eugenol, acetovanillone, etc.):

1) The use of WB-MIX avoids production problems involved in the use of spices in the brewhouse:
– microbiological problems due to the presence of a whole series of exotic microorganisms on spices ;
– aromatic character variations from crop to crop and from one provider to another ;
– deterioration of spices aromatic properties during storage ;
– extract yield variations of spices aromatic compounds in the boiling kettle.

2) WB-MIX use avoids production problems related to the use of a different yeast strain to produce the metabolites that impart the typical white beer taste since similar compounds to these typical metabolites are found in the specific spices contained in WB MIX.

3) The use of WB-MIX gives a higher taste reproducibility because all the aromatic compounds of the mixture coming from spice CO2 extracts are standardized so as to have always the same concentration in this mixture.

4) High reproducibility of beer turbid aspect.

5) New white beer production start-up can be done very easily since we can use as base beer a beer already produced in the brewery (the unique requisite being a low bitterness: 10-12 EBU).

6) The use of WB-MIX offers high production flexibility allowing a fast response to the market.

7) All the ingredients of WB-MIX are 100% natural and can be mentioned on the label of beer like vegetable extracts or natural flavours or spices or beer aromatized with spices (Benelux AR 31 March 93 beer Article 4 § 2).

8) This production way allows to produce a Belgian type white beer without alcohol (0,5% v/v).

WB-MIX is a composition and not the result of a distillation from which we can not control the components individually. The spices are extracted with CO2 and are standardized with regard to most representative component of each spice. The concentration of one spice can be modulated precisely independently from other spices. This allows to obtain an excellent reproducibility of the WB-MIX formula.

WB-MIX is the result of a market survey, a marketing strategy, fundamental search on flavours (beers and spices), adjustment of search results to the consumer (tastings) and the follow-up of beers produced by this way since 1997.

Results of chromatographic analysis of 60 commercial white beers :

Values in ppm
mean
60 beers
minimum
60 beers
maximum
60 beers
flavour/
taste
propanol-1
isobutanol
isoamyl alcool
2-phenyl alcool
ethyl acetate
isoamylacetate
ethylcaproate
ethylcaprylate
ethylcaprate
phenylethylacetate
18,80
36,50
58,00
33,00
32,00
4,00
0,12
0,25
0,05
0,98
11,30
19,80
41,00
13,00
13,00
0,70
0,06
0,04
0,01
0,09
52,10
59,80
102,00
64,00
54,00
9,00
0,20
0,37
0,26
2,24
fusel
fusel
fusel
rose
solv/fruity
very fruity
fruity/wine
fruity/wine
fruity/wine
rose/honey
Values in ppb
mean
60 beers
minimum
60 beers
maximum
60 beers
flavour/
taste
phenol4
guaïacol
4-vinyl phenol
4-vinyl guaïacol
eugenol
isoeugenol
4-vinyl syringol
4-OH-benzaldehyde
acetovanillone
vanillin
40
120
970
1500
70
38
310
125
153
110


10
280


96


110
360
7390
3710
215
156
586
318
474
450
phenolic
phenolic
phenolic
cloves
phenolic
cloves
smoke
phenolic
vanilla
vanilla

 

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