2013-11705-01 Brewing Handbook Final Spreads
2013-11705-01 Brewing Handbook Final Spreads
2013-11705-01 Brewing Handbook Final Spreads
Brewing
Handbook
Version 1 2013
* Adjunct silo = Unmalted cereals (barley, wheat, maize (corn), rice etc.
table of contents
Foreword
introduction
77
70
80
82
10
11
Chapter 6
12
Attenuation Control
12
85
14
86
87
87
Chapter 2
raw material optimization part 1
17
88
18
90
20
92
95
21
26
Chapter 7
33
Fermentation control
with FAN optimization
Chapter 3
99
38
40
100
101
101
37
100
102
103
105
Chapter 4
cost-effective cereal cooking
51
Chapter 8
52
Diacetyl control
107
54
108
54
108
55
109
59
110
60
112
62
115
117
References
121
Chapter 5
Efficient wort separation
and beer filtration
65
66
67
68
foreword
Foreword
Over 30 years ago Novo Nordisk A/S (today Novozymes) introduced industrial,
microbially produced enzymes for the brewing industry. The first products
were a bacterial protease and a bacterial alpha-amylase. Our offering for the
brewing industry has since evolved into a comprehensive portfolio of enzymes
combined with an extensive range of services to meet your needs, whether it
is optimizing your products and production processes or developing innovative
new products. Technical information on these enzyme products and how
they can be used in brewing is available in a number of separate information
brochures, lectures, and published articles. The Brewing Handbook brings the
most relevant information together in one single publication for easier access
and reference.
The purpose of this publication is to support breweries to improving the beer to
improving the production economy, process control or beer quality.
Our belief is that quality solutions require both the product and the service
to be outstanding. In line with globalization and the trend for customizing
solutions, the demand for great service is steadily growing. And as that demand
grows, our support is growing to ensure that we can continue to meet the
needs of the brewing industry and we see this handbook as an integral part
of our support for the brewing industry.
chapter 1. introduction
Chapter1. introduction
Chapter 1.
introduction
chapter 1. introduction
sustainability.
needs. We can also help secure right-first-time processes with a variety of raw
enzymes are a tool for breweries to reach their strategic business goals.
materials, and ensure the most profitable route to your high quality beer.
Together we can unlock opportunities that secure the future of your brewing
business.
your processes are right the first time and assist you in finding excellent new
ways to utilize your capacity. At the same time, we can help you save energy
tool. For example, enzymes make it possible to utilize local raw materials,
background and mode of action of each solution, and provides practical advice
which can not only reduce your input costs but can also support the local
and real examples. We hope it will become an invaluable aid for you when
economy. Enzymes give you the flexibility to rethink the brewing process,
brewing!
10
11
chapter 1. introduction
consistency are an integral part of who we are. We have systems for assessing
Ondea Pro or Novozymes Ceremix, you can instantly achieve excellent raw
and approving suppliers, and our IT systems ensure traceability of our products
material flexibility and more sourcing options through benefiting from brewing
from supplier to you. Our long tradition of working actively with health and safety
with alternative raw materials. Depending on your brewerys location and local
issues ensures that our products are safe to use and safe to handle. We use safe
raw material availability, you could have the option of sourcing barley, cassava,
sorghum etc. Not only does this deliver cost savings, it also enables you to
of our enzymes which minimize any risk. We have acquired our safety expertise
through decades of producing enzymes and share it with our customers through
safety and warning labels and Material Safety Data Sheets (MSDS).
Our global business is covered by ISO 9001, and we also hold the ISO 22000,
FSSC and AIB certifications for plants producing a wide range of food and
beverage enzymes, including internationally recognized kosher and halal
compliant products. Our solutions are approved by all relevant authorities and
international committees.
As brewing is a sensory business where consumers judge beer one serving at a
time, we ensure the consistent quality of your brands by producing the majority
of our brewing enzymes in compliance with BrewQ specifications. This means that
they are additionally analyzed according to Analytica-Microbiologica-EBC 2005
Section 4.6.1.
12
13
chapter 1. introduction
support you in implementing and optimizing our solutions to fit your needs,
Denmark, Switzerland, Russia, South Africa, Malaysia, USA, India, Japan and
China; bases from which trial support and application recommendations can be
offered. Our unique global distribution set-up secures product availability in any
location. Were looking forward to working with you to meet the future needs
Production
R&D
Sales offices
Global marketing
14
15
Chapter 2.
17
Key benefits
expectations have always influenced the selection of the brewing raw materials.
Achieve faster and advanced viscosity reduction and increased extract yield
However, increasing cost pressure in the industry has led to further constrained
adjustments in beer recipes over the last couple of years, with more focus on
cost effective and sustainable alternatives. The industry is also challenged by
seasonal and regional availability, fluctuation in price and quality caused by
climatic conditions during cultivation and harvest. As a consequence, there is
processability, increase yield and assure wort and beer specifications. Broadly
compensate for variability as well as fluctuations in the grain market and raw
material quality.
Novozymes Ultraflo
Optimize your processability, starch degradation and FAN release with
Ceremix
Novozymes products are developed to work either in synergy with the existing
enzyme systems in the various grains (barley, malted barley, wheat etc.), or
increase of maltose
18
19
malt and to increase the extract yield by approximately 1%, Ultraflo Max is
compromising yield. If, on top of the raw material optimization, the Real Degree
of Fermentatiion (RDF) specification is increased, a dosage of 0.05 to 0.1 kg of
Attenuzyme Pro per ton of grist enables an RDF increase by approx. +1% (Max
72-74% RDF).
Rice/
Malted
Barley Wheat Maize Sorghum
barley
(corn)
100
80
Ultraflo
Max
Ceremix
Plus MG
Ondea
Pro
Termamyl Attenuzyme
SC DS
Pro
Neutrase
1.6 L BrewQ
0.10-0.15
20
0.12-0.18
0.10-0.25
80-100% Malt
Optional
Ultrao Max
60
40
0.12-0.18
0.25-0.60
40
60
0.6-1.2
20
80
1.2-1.8
100
1.8-2.2
0.10-0.35
Optional
80
20
0.12-0.15
0.17-0.25*
60
40
0.12-0.15
0.17-0.25*
80
20
0.12-0.18
40
0.15-0.25
0.25-0.70
40
60
0.15-0.25
Fig. 2.2-1. Enzyme recommendation for malt-based recipes with minor barley inclusion
0.10-0.25
60
0.25-0.50
1.2-1.5
60
40
0.17-0.25*
20
80
0.17-0.25*
50
20
30
30
50
20
30
40
30
0.12-0.18
0.25-0.50
0.20-0.50
or Fungamyl BrewQ
0.5-1.0 kg/ton
0.20-0.50
or Fungamyl BrewQ
0.5-1.0 kg/ton
0.17-0.25*
0.6-1.2
0.17-0.25*
1.2-1.8
0.15-0.30*
Raw materials:
85% malted barley
+ 15% raw barley
Ceremix Plus MG
(kg/ton grist)
Ultraflo Max
(kg/ton grist)
Attenuzyme Pro
(kg/ton grist)
Reference
Application example
0.10
0.12
0.05
Table 2.2-1. Example of effective enzyme treatment on 85% under modified malt and 15% barley
20
21
Raw materials:
85% malted barley
+ 15% raw barley
Filtration
(ml/10)
Extract
(P)
-Glucan
(16.0P)
FAN
(16.0P)
Viscosity
(16.0P)
Reference
43
15.86
1049
192
Application example
67
17.15
139
213
DP 1
(%)
DP 2
(%)
DP 3
(%)
DP 4/DP4+
(%)
Ferment-ables
(%)
Expected
RDF
2.516
17.8
42.5
13.9
25.8
74.2
66.8
1.951
22.6
42.4
13.5
21.5
78.5
70.7
MG per ton of malt compensates a lack in malt modification and assures high
processability and fermentability.
0-30% Barley
40-100% Malt
Table 2.2-2. Results of wort analysis after applying Novozymes Ultraflo Max, Novozymes Ceremix
Plus MG and Novozymes Attenuzyme Pro on 85% under-modified malt and 15% barley
Ultrao Max
Termamyl SC DS
Processing high gelatinizing adjuncts like maize (corn) and rice in a cereal cooker
Ceremix Plus MG
0.25-0.70 kg/ton barley
Fig. 2.2-3. Enzyme recommendations for malt-based recipes with higher portions of alternative raw
you can achieve high processability and a very robust brewing set-up.
70-100% Malt
Using the full potential of exogenous enzymes you can create recipes with
Ultrao Max
Termamyl SC DS
up to 100% barley. However, any ratio of barley, wheat and malt can be
processed efficiently. Ondea Pro enables brewers to brew maltose-based wort
with standard fermentability and similar processability compared to using
high portions of malt. The present pullulanase, amylase and protease activities
in Ondea Pro ensures sufficient starch and protein degradation in synergy
with the -amylase and peptidases of the barley. The glucanase and xylanase
components enable sufficient cell wall degradation and low viscosity. The lipase
activity significantly improves the turbidity during lautering.
Fig. 2.2-2. Enzyme recommendation for malt based recipes including rice or maize (corn).
60-100% Barley
0- 40% Malt
Ondea Pro
22
23
30-60% Barley
to 2.2 kg of Ondea Pro per ton of barley. Tables 2.2-3 and 2.2-4 show three
30-60% Malt
20-40% Wheat,
rye, oat, triticale
different recipes that use the full potential of Ondea Pro for highly cost-
Ultrao Max
Attenuyzme Pro
Barley
(%)
Wheat
(%)
Malted barley
(%)
Ondea Pro
(kg/ton of grist)
Ceremix Plus MG
0.50 kg/ton total grist
Application
example A
100
2000
Application
example B
50
35
15
35
50
15
1500
Application
example C
Ondea Pro
1500
Fig. 2.2-5. Example of how to use Novozymes enzymes to utilize individual raw material agenda
Additionally, Novozymes can provide ready to use solutions for cassava and
Table 2.2-3. Different recipes using the full potential of Novozymes Ondea Pro for highly cost-
Application
example A
42
Extract
(P)
Turbidity
(NTU)
15.03
-Glucan
(16.0P)
12
64
ar-Xylan
(16.0P)
215
Viscosity
(16.0P)
1.942
DP 1
(%)
8.8
DP 2
(%)
46.7
DP 3
(%)
18.5
DP 4/
DP4+
(%)
26.0
Fermentables
(%)
Expected
RDF
Recommended
products
74.0
66.6
Application
example B
55
15.67
14
56
236
1.937
10.0
50.4
17.2
22.4
77.6
69.9
Application
example C
54
15.88
17
58
240
1.944
9.5
53.1
17.1
20.2
79.8
71.8
Termamyl SC DS
-glucanase
Xylanase
-amylase
Protease
-glucanase
Xylanase
-amylase
Pullulanase
Protease
Lipase
Attenuzyme Pro
Glucoamylase
Pullulanase
Neutrase 1.6 L
FAN optimization
Better starch degradation
Protease
Ceremix Plus MG
aw material agenda
Ondea Pro
-amylase
-glucanase
Xylanase
Ultraflo Max
Main enzyme
activities
Table 2.2-4. Results of wort analysis after applying Novozymes Ondea Pro on mixes of barley,
wheat and malt
Benefits
24
25
-glucan degradation
-glucan is a polysaccharide composed of D-glucose molecules with -1,3
To seize the cost saving opportunities that come with alternative raw materials
based on it, can be the limiting factor. Either the enzymes are not sufficient in
terms of temperature or pH characteristics, or the amount and function do not
CH2OH
support the set-up of a modern raw material agenda. The following section
describes the different enzyme systems used in brewing to fulfill the required
processability and fermentability, and to reach the target quality specifications.
CH2OH
CH2OH
O O
OH
-1,4
The husk of barley and barley malt contains approximately 5-6% cellulose
OH
OH
O O
-1,4
HO
-1,3
O O
OH
OH
-1,4
OH
O O
OH
Endo-glucanase, cellulase
n
Endo--1,3(4)-glucanase
cell walls in the endosperm consists of approximately 65% -glucan and 25%
pentosans. Both substances are critical to the brewing process in terms of starch
During the brewing process -glucan with high water binding capacity is
released. If not degraded sufficiently, -glucan causes high wort and beer
using exogenous -glucanases and GH-10 family xylanases. Fig. 2.3-1 shows the
structure of the cell walls linked together by proteins in the middle lamella.
decreased mash and beer filtration performance. Amongst others long chain
-glucan underlies stretching during the process, for instance in pumps, which
can result in additional windings into micelles that punctiliously block the
Ferulic acid
Acetic acid
filtration steps.
Arabinose
The relevant -glucan degrading enzymes are the -glucan-solubilase, the endoand exo--glucanases as well as cellulases. The -glucan-solubilase belongs to
the enzyme class of esterase and dissociates the high molecular hemicellulose
Arabinoxylan
-glucan
Arabinoxylan
-glucan from the proteins in the cell wall matrix. The optimum temperature of
the endogenous -glucan-solubilase is around 62-65C and it is deactivated at
Protein
72-73C.
Carboxypeptidase
Ferulic acid
Arabinoxylan
-glucan
Arabinoxylan
Protein
-glucan
Arabinoxylan
Arabinoxylan
Cell wall 1
Middle lamella
Cell wall 2
-glucanases cut the -glucan from the non-reducing end and form cellobiose.
This reaction reduces the viscosity slowly.
26
27
temperatures above 40C and therefore not relevant under normal brewing
of fermentation
conditions.
The primary focus of the brewing process is starch conversion into fermentable
sugar and dextrin. The amount of extract released from degradation of mainly
starch and the final degree of fermentation forms the basis for the produced
and pH optima indicate that the in-malt cytolytic enzyme system does in most
cases not have the optimal properties for the brewhouse process working at a
brewing, the starch hydrolysis are mainly transformed by the and -amylases.
While in unmalted conditions, the -amylase is already sufficiently present in
Like the -amylase, some of the cytolytic enzymes are not present in barley
most cereals like barley, wheat and sorghum, the -amylase is formed de novo
and are formed during the malting process. Before malting the barley contains
Amylose
more critical -glucan at higher temperatures during mashing. Also the exo-glucanase activity is decupled during malting. The increase of the exo-glucanase, however, depends on the variety and climate conditions.
CH2OH
OH
CH2OH
OH
CH2OH
O
O
OH
Amylopectin
CH2OH
O O
OH
OH
OH
-D(1 6) Bond
O
OH
OH
CH2OH
-D(1 4) Bond
and advanced viscosity reduction results in high performing mash and beer
CH2
O
O
OH
OH
-D(1 4) Bond
CH2OH
O
O
OH
O O
OH
OH
OH
filtration.
Fig. 2.3-3. Chemical structure of amylose and amylopectin
Arabinoxylan degradation
Similar to the -glucan, the pentosans, especially the arabionoxylans,
significantly impact the wort and beer viscosity and the performance in mash
amylose and amylopectin from the inside. The major products are dextrins.
and beer filtration. The barley contains an analogue to the glucanases prevalent
However, with increasing mashing time, the -amylase can degrade the
of the -amylase and partly below the gelatinization temperature of maize (corn),
and reduces the wort viscosity within an intensive mash regime. However, the
temperature optimum is around 45C, making the activity nearly irrelevant for
modern brewing conditions. The exo-xylanase degrades the xylan from the end,
but only if the substrate was already released because of the endo-activity. The
-amylase degrades the amylose from the non-reducing end and cuts off
remaining in-cereal cytolytic activities are limited and the activity increase minor
maltose. If the glucose chain is unequal, the last three glucose units are not
attacked and stay in the wort as maltotriose. The temperature and pH optima
of the -amylase under brewing conditions are 60-65C and pH 5.6-5.8.
to degrade all the dextrin into fermentable sugars. Even with a highly modified,
filtration, boost the beer filtration both in kieselguhr and membrane filtration
enzyme rich malt and intensive mashing, the real degree of fermentation is
systems.
28
29
decisive for using either the infusion or decoction process. Raw barley and
like the Novozym 26062 can both increase the amount of maltose in wort in
wheat, as well as triticale, oat and rye, have a similar gelatinization temperature
synergy with the cereal -amylase and speed up the saccharification process
However maize (corn), rice, sorghum and cassava need to be gelatinized and
liquefied at higher temperatures in a separate cereal cooking process.
Table 2.3-1 shows the gelatinization temperatures of the common brewing raw
Raw
material
Barley Barley
malt
Wheat Maize/
corn
Rice
Sorghum Cassava
the free amino nitrogen (FAN) content becomes critical even if an advanced
yeast management system is in place. However, the proteolytic degradation
during malting and mashing not only releases amino acids and dipeptides as
Gelatinization
temperature
(C)
yeast nutrients, but also enables and support access to starch. The protein
60-65
61-65
55-65
64-82
68-84
68-75
64-76
in the endosperm is linked to the -glucan and pentosans in the cell walls
surrounding the starch kernels. This becomes most relevant for high protein
wheat and especially sorghum processing, which has large amounts of kafarin
in the endosperm.
of the malt loading in the decoction step, conducting an intensive rest at 72C
oligopeptides from the inside, the exo-peptidases are responsible for releasing
single amino acids and dipeptides. Fig. 2.3-4 shows the principle in protein
Exogenous enzymes can certainly support and partly substitute the malt-based
hydrolysis.
Endopeptidase
Exopeptidase
Dipeptidase
step and in the mashing process. Because of specific screening for enzymes
with an activity optima relevant for brewing, these heat-stable amylases
Albumin
Protein
Globulin
Prolamin
Glutelin
even lead to a faster viscosity break and yield increase. The properties of the
exogenous enzymes also provide the opportunity to lower the maximum
substitutable with exogenous enzymes because the activity in the cereal is more
than sufficient. However, the exogenous enzyme tool box can add functionality.
y p e p ti d a
se
Oligoopeptide
rb o x
Polypeptide
Ca
opeptidase
Amin
Endopeptidase
Macropeptide
Tripeptide
Dipeptide
Aminoacids
solutions help increase and control the RDF beyond the malt-based limits
for production of light or strong beer. In fact, the resultant wort is based on
30
31
while the minor part is activated by metal. The individual peptidases work
specifically on certain amino acid bonds. Partly the endo-peptidase activity
enzymes are more heat-stable than the malt glucanases. This allows sufficient
Glucose
proteins from the end of the free amino group. While the carboxypeptidase
activity is increased during malting, the aminopeptidases are to a large extent
-1,3-
-1,4-1,4-
-1,4-1,4-
-glucosidase
Endo -1,3(4)-glucanase
protein rest during mashing. Both methods, however, have often shown to
be insufficient to give an acceptable FAN level when using high amounts of
adjuncts. Novozymes Neutrase products are working in synergy with the
Xylose
Arabinose
Endo-xylanase (GH11)
Exo -xylanase
Ferulic acid
Acetyl
Glucoronic acid
Endo-xylanase (GH10)
-glucoronidase
32
33
AG
Amyloglucosidase
AG
-amylase
-A
-A
P
-amylase
Pullulanase
-A
-A
Endo-protease
C-terminus
N-terminus
Exo-protease
Fig. 2.4-4. Protein structure and the effect of endo and exo-proteases
34
35
Chapter 3.
37
Beside the major brewing raw materials barley and barley malt, various starch
sources like maize (corn), rice, wheat, sorghum, rye and cassava, as well as
Rye
Triticale
3% 3%
3%
Sorghum
Barley
Europe
Rice
4%
3%
6%
Barley
19%
syrups and sucrose from both sugar cane and sugar beet, are widely used in
Wheat
51%
the brewing industry. Raw material crops are handled on a global trade market.
Wheat
18%
70%
Americas
22%
Maize (corn)
Maize (corn)
scale, the barley crop of approximately 125 million MT p.a. accounts for only
5% of the global production of relevant grains. Fig. 3.0-1. shows the global
Millet
6%
Rice
Barley
2%
Cassava
Africa
6%
Barley
2%
Asia
8%
Barley
Sorghum
Sorghum
20%
45%
Raw material
5% 2%
Cassava
Maize (corn)
8%
9%
Wheat
32%
Cassava
49%
Rice
8%
Maize (corn)
25%
Wheat
27%
Rice, paddy
Raw
Material
Maize (corn)
Rice
Wheat
Cassava
Barley
Sorghum
Production in
Mio. MT (2010)
840
700
650
230
120
60
24%
24%
Maize (corn)
Wheat
With 120 million MT, Africa represents approx. 50% of the global production
of cassava and this accounts for 45% of the starch source produced on the
African continent. Other major producers of cassava include USA and India.
Furthermore, maize (corn), sorghum, millet, wheat, rice and smaller quantities
of barley are relevant in Africa to cover the demand for food, feed and
These grains are mainly used for food, feed and partly for the production
beverage industries. Various crops like barley, rice and sorghum are mostly
of bioethanol. Sugar cane and sugar beet are not displayed. However, with
Nigeria, Ethiopia and in the Sudan area, whereas barely is grown in significant
globally. While sugar beet is also grown in Europe, sugar cane is mainly planted
crop is maize (corn). Rice is the most important food source in Asia followed by
wheat and maize (corn).
The cost of rice has increased significantly over the last decade. Rice has been
while the Americas, and in particular USA, grow maize (corn) in significant
used in large quantities for beer production. However, cost pressure has led
amounts.
producers to look for alternative brewing materials. Even though cassava and
barley play a minor role in the Asian agriculture sector, they have become
more in focus in the brewing supply chain. To cover the demand of the Asian
brewing industry, Oceania and Europe are important sources of barley, barley
malt and wheat which are all imported in a significant amount.
38
39
Despite the large amounts of adjuncts in beer recipes in Asia the brewing
industry has to deal with a regional undersupply and needs to import barley
The barley malt trade market reflects the malting barley agricultural situation.
illustrates the ten largest barley producers globally, with a production of more
Traditional barley growing countries also have major malting capacity. Main
exporters are France, Canada, Australia and Belgium. In contrast, Brazil and
two-thirds of the worldwide barley production. Fig. 3.1-1 shows that Europe
Japan are the main importers of malt. This is demonstrated in Fig. 3.1-3.
is the largest producer of barley. Together with Canada and Australia, Europe
represents the heart of the global barley supply for the brewing industry.
Russia
Spain
-0.4
-0.9
Viet Nam
Uruguay
Ukraine
United Kingdom
Sweden
Thailand
Slovakia
South Africa
Russian Federation
Romania
Poland
Nigeria
Philippines
Mexico
Netherlands
Italy
Hungary
France
Germany
Finland
Chile
China
Canada
10%
10%
Canada
Ukraine
-0.9
Republic of Korea
11%
9%
-0.4
Japan
9%
0.1
Czech Republic
Australia
13% France
0.1
Cameroon
9%
0.6
Belgium
Turkey
0.6
Brazil
barley-producing
countries
Austria
10 biggest
Belarus
7%
Germany
13%
1.1
Australia
5%
1.1
Argentina
United Kingdom
USA
Malt
Fig. 3.1-3 also shows that China is neither importing high quantities of malt nor
growing sufficient amounts of barley. Furthermore, the local barley crop might
Malting barley
The difference between barley growing countries and main malting barley
suppliers is marginal. Still Europe is accountable for approximately 44% of
The Chinese malt supply is not directly linked with malt imports, but malting is
the worldwide supply see: Fig. 3.1-2. South America, with 13%, has a larger
share of the malting barley market although it is not among the top 10 barley
growing nations. The largest beer market, Asia, is only producing approximately
due to the fact that the barley husk can be used as a natural filter cake for the
Russia/Ukraine
Asia
6%
South
America
Africa
Malting barley
5% 1%
Starch
13%
44% Europe
Oceania
Barley composition*
Protein
Pentosans &
-glucans
Cellulose
Lipids
Ash
69-73%
9-13%
10%
5-6%
2.5%
3%
14%
Table 3.1-1. Average composition of brewing barley (*on dry matter)
17%
USA
40
41
Therefore the starch content per ton of traded material is higher and the
and produced malt belong to the fraction of low gelatinizing starch sources. In
combination with the natural enzyme system which is present in the raw barley,
and additionally produced during malting, these properties are the foundation
for the common temperature profile in an infusion mashing process.
Wheat composition*
Starch
Protein
Pentosans &
-glucans
Cellulose
Lipids
Ash
72-77%
11-15%
7-8%
2-3%
2%
2%
With more than 635 million MT p.a., wheat is the third largest global grain
crop. The wheat crop shows an opponent agricultural distribution in terms of
major growing areas. In the wheat market, the ten biggest wheat producers
are accountable for more than 450 million MT, growing approximately 70%
of the crop worldwide. Asia, and in particular China and India, with their
high population, play an important role in global wheat production. This is
Rice, maize (corn) and sorghum belong to the high temperature gelatinizing
Australia
Canada
Turkey
5%
China
5%
Pakistan
10 biggest
4%
25%
wheat-producing
countries
planting it. Compared to wheat or barley, the yield is nearly double and has
increased significantly over the last few years, indicating that grain breeders
5%
Germany
million MT. This is not only due to the very high yields farmers can achieve by
have an enormous focus on this raw material. Maize (corn) is also the raw
5%
9%
17%
9%
Russia
crop distribution of the 10 biggest maize (corn) producers globally who are
India
13%
USA
accountable for approximately 80% of global maize (corn) production; see Fig.
3.1-5. Out of these countries, USA is harvesting 47% followed by China and
Brazil with 26% and 8%. In Europe, France is the largest maize (corn) producer,
but only contributing to 1.5% of global production.
To plant and breed wheat for brewing is not a local point of the agricultural
agenda. The amount of wheat that is used in brewing is marginal compared to
the food sector. The main challenge of insourcing wheat for beer production
is the food industrys deviating focus on protein levels. While for the food
industry, high protein content is equal with high, first grade quality, brewers are
South Africa
Ukraine
France
India
2%
Indonesia
2%
3%
Argentina
3%
Mexico
10 biggest
maize (corn)-producing
countries
3%
Brazil
47% USA
8%
looking for wheat with less proteins which in the sense of food production,
is not first grade quality. However, this is an opportunity for economically
attractive sourcing on the regular wheat market. Wheat has a similar
26%
China
42
43
Table 3.1-4 displays an average composition of rice. Similar to maize (corn), rice
protein is not accessible during mashing and the nitrogen nutrients need to be
minimize metabolic losses, equal to other cereals. The high lipid content of
maize (corn) can impact the beer quality negatively in terms of foam and flavor
stability. Most of the oil is located in the embryo. So for brewing, the maize
Rice composition*
Starch
Protein
Pentosans &
-glucans
Cellulose
Lipids
Ash
84-88%
5-9%
2%
2.0-2.5%
0.5%
0.5%
Protein
Pentosans &
-glucans
Cellulose
Lipids
Ash
73-77%
8-11%
5-6%
4%
5-6%
1.5%
Sorghum is cultivated in warm climates. For the brewing industry, you mainly
find this in Africa. Nevertheless, the biggest producers of Sorghum are USA,
Mexico and India, Fig. 3.1-7. As for Africa, this genus of grass species is
mainly grown in Nigeria, Ethiopia and Sudan and is greatly important for beer
The protein content of maize (corn) is not significantly accessible during
production there. Sorghum accounts for only 2% of the worldwide grain crop
mashing and it doesnt contribute to the nitrogen supply of the yeast during
Sorghum can generally be separated in two groups: the white sorghum and
the brewing process. That makes the amount of corn in brewing recipes limited
to 50-60%. In breweries, maize (corn) can be used as corn grits, flakes, pre-
of taste and quality. White sorghum is used for malting or directly for beer
production. Pure sorghum beers are produced in Africa. The gelatinization
Rice is the most widely consumed staple food for a large part of the worlds
population, especially in Asia and the West Indies. Worldwide rice production is
close to 700 million MT p.a. More than 85% of annual production is grown by
the 10 biggest producers.The major contributors in the Asian region are China
China
Australia
and India (see Fig. 3.1-6). The next largest producers are Indonesia, Vietnam
and Myanmar. The only two non-Asian countries in the top ten are Brazil and
5%
Sudan
3%
20%
Ethiopia
sorghum-producing
countries
7%
Argentina
8%
with the highest gelatinization temperature; up to 85C. However, rice also has
the highest naturally occurring starch content; 84-88%.
10 biggest
USA
6%
USA, accountable for less than 3.5% of global rice crops. Alongside food
production, broken rice is usually used for beer production. Rice is the adjunct
4%
Burkina Faso
16%
Mexico
11%
Nigeria
Brazil USA
Philippines
Thailand
3% 2%
5%
Myanmar
10 biggest
2%
5%
Vietnam
32%
China
15%
India
rice-producing
countries
Fig. 3.1-7. Sorghum crop distribution (2010)
6%
The 11-12% protein in sorghum and sorghum malt can be solubilized during
Bangladesh 8%
11%
Indonesia
24%
India
44
45
Sorghum composition*
This makes local, sustainable sourcing an opportunity for either specific brands,
Starch
Protein
Pentosans &
-glucans
Cellulose
Lipids
Ash
78-80%
11-12%
2%
6.8%
3.7%
1.5%
Sweden
Spain
China
5%
Australia
Belarus
starch in cereal cooking. The starch content is comparable with barley, but the
Canada
7%
8%
16%
8%
France
Poland
20%
6%
10 biggest
4% 2%
Germany
15%
Russia
Grain
-amylase*
-amylase*
Barley
0.62
350
Cassava
nd
nd
Today, oats are mainly used for animal feed and breakfast cereals, while rye and
Corn
nd
nd
triticale are already widely used in distilling and the production of bioethanol.
Malted barley
280
920
Rice
0.12
57.4
Sorghum
0.36
252
Wheat
0.42
454
Oat composition*
Starch
Protein
Pentosans &
-glucans
Cellulose
Lipids
Ash
72-76%
12-14%
5-6%
4-5%
7%
3%
Oats, rye and triticale belong to the so called secondary crops that are not yet
in the focus of the brewing industry. However, these grains are today used
Rye composition*
Starch
Protein
Pentosans &
-glucans
Cellulose
Lipids
Ash
72-74%
11-14%
6-7%
5-6%
2%
1.5%
for some special beer brands that use the properties of these raw materials to
position the beer with healthy attributes. Oat and rye can be traced back to the
Stone Age and are well known for their modesty in terms of soil and weather.
These crops obtain reasonable yields even in cooler regions. However, triticale
is a hybrid based on rye and wheat. Breeders combined the modesty of rye
Triticale composition*
Starch
Protein
Pentosans &
-glucans
Cellulose
Lipids
Ash
68-72%
11-13%
8-9%
4-5%
1-2%
2.1%
with the agriculture yield and quality of wheat. In recent years, a considerable
amount of new triticale varieties has been evaluated and registered.
Based on their characteristics oats, rye and triticale could be utilized as raw
Table 3.1-7. Average composition of oat, rye and triticale (*on dry matter)
materials for brewing, especially in Northern and Eastern Europe. Fig. 3.1-8
shows that the crop is actually dominated by Poland, Germany and Russia
The Cassava root can be seen as the rising star of raw material for brewing due
which are accountable for 50% of the global production. France, Canada,
Australia and Belarus are also growing a significant amount of these grains.
the ability to support brewing groups reaching their social sustainability targets.
Cassava can support production for low cost segments, or replace expensive
sugar or syrups in all beer segments.
46
47
Cassava accounts for 45% of the relevant crops in Africa and 6% of the
unpublished data 2013, in the table below, is also very valuable when deciding
In the case of cassava, global production does not mirror the global trade
market. Thailand is the dominant supplier to world markets accounting for
approximately 80% of global trade. Vietnam, Indonesia and a few countries in
Cellulose
Africa and Latin America share the remaining market. This situation is mainly
Non-cellulosic
polymers
Pectic
polysaccharides
India
Vietnam
5%
Polygalacturonic acids,
which may be substituted
with arabinan, galactan
and arabinogalactan
Mozambique
5%
3%
10 biggest
Nigeria
cassava-producing
countries
21%
Chips
Pellets
Pulp
751.4
678.3
373.5
Amylose (g/kg)
173.6
180.2
113.2
Amylopectin (g/kg)
578.2
497.6
260.9
Amylose / Amylopectin
0.29
0.36
0.43
389.7
310.8
592.1
18.89
25.69
12.96
Angola
Parameters
Total starch (g/kg)
Ghana
8.28
8.27
27.90
42.19
53.30
97.37
8%
8%
14%
Congo
Brazil
8%
12%
13%
Indonesia
Thailand
Cassava is already widely used within many large industries in food, feed
Table 3.1-9. The carbohydrate contents and property of starch granule for the cassava products
start straight after the harvest to avoid rotting. Processed starch or cake can
then easily be integrated into the brewing supply chain. The gelatinization
temperature is slightly higher than that of barley. Therefore, the cassava needs
to be liquefied in a cereal cooking step beforehand. Table 3.1-8 shows a typical
composition of cassava chips.
Cassava chips*
Starch
Protein
Pentosans &
-glucans
Cellulose
Lipids
Ash
86-90%
3-5%
1.1%
2.8%
1.3%
1.5%
48
49
Chapter 4.
50
51
This is of course very dependent on the actual starch type and quality. Typically,
temperatures between 85C and 100C are used in the cereal cooker. At these
Starch containing adjuncts and cereals must be processed in such a way that
stable -amylases are frequently used in brewing with these types of adjuncts.
down the intermolecular bonds of starch; both amylose and amylopectin, in the
presence of excess water and heat in order to engage more water, also known
Key Benefits
Low viscosity
149 F 65 C
saccharification
Reduced processing costs through more efficient liquefaction and increased
yield of up to 1%
Gelatinization of starch
185 F 85 C
Enzymatic degradation of starch
High viscosity
Fig. 4.0-1. The gelatinization and liquefaction process a schematic approach. After gelatinization,
the viscosity is lowered due to the action of -amylases resulting in an enzymatic degradation of
starch, also known as liquefaction
52
53
Termamyl is added to the cereal cooker with the adjunct at the start of
amylase, which breaks down the -1,4- linkages in amylose and amylopectin
glucose linkages and is thermostable. The -1,6- linkages are bypassed and are
not hydrolysed. This enzyme also reduces the viscosity of starch suspensions
2+
and produces dextrins which are compounds that contain up to twelve glucose
2+
units. -amylase is largely absent from unmalted barley and wheat. Review
the raw material optimization section on page 46. Table 3.1-6, for further
information.
the viscosity of the resulting wort. If the chains remain long, the chance of
retrogradation is higher. The retrograded starch precipitate represents a loss in
Gelatinization temperature
To be mashed in the
mash tun
cereal cooker
Barley
60-65
140-150
Cassava
64-76
147-169
Maize (corn)
64-82
147-180
Oat
53-60
127-140
Rice
68-84
154-183
Rye
57-70
135-158
Sorghum
68-75
154-167
Triticale
61-64
142-147
Wheat
55-65
131-149
Slow
Solution
Rapid
Precipitate
Gel
x
x
x
x
Table 4.2-1. Gelatinization temperatures of common brewing cereal grains; average values and
process
54
55
90
Temperature (C)
gives the mashing/time profile, in the cereal cooker, as shown in Fig. 4.3-2.
100
110
70
60
50
40
100
Temperature (C)
80
90
25
50
75
100
Time (minutes)
80
70
60
50
40
0
30
60
90
120
150
Time (minutes)
Fig. 4.3-3 is showing a normal mashing regime in the cereal cooker with
the addition of Termamyl instead of malted barley. In Fig. 4.3-4 a normal
viscosity graph is shown during the cereal cooking process with a peak during
gelatinization.
Due to the low heat stability of the malt -amylase, relatively high viscosities
Viscosity
Temperature (C)
very quickly during this process and are lost for later utilization during mashing
and mash filtration.
6000
Viscosity (cP)
7000
Maximum viscosity
After cooling
100
5000
80
4000
3000
Liquefaction viscosity
2000
1000
120
60
40
20
Temperature (C)
if malt is used for adjunct liquefaction, all the other enzymes (-amylase,
Fig. 4.3-4. demonstrates the viscosities during a cereal cooking step after the addition of
Novozymes Termamyl
Using Termamyl, the cereal mash viscosity is greatly reduced, thereby preventing
formation of retrograded starch. This means that the yield from using adjuncts
times higher per kg than that of malt. Thicker mashes can be operated without
the risk of working with high viscosities. This, in combination with the fact
The yield can be increased by more than 1%. While it is necessary for malt
that 100 kg of malt is replaced with 0.19 kg Termamyl SC DS, enables smaller
-amylase to work at its limit for temperature stability, Termamyl maintains very
working with high proportions of adjuncts. This versatility can also be used to
increase brewhouse capacity. In addition to what is achieved by working with
thicker mashes, the malt is replaced with adjuncts with higher extract values.
56
57
The process is also a more cost effective one, as all the malt can be utilized
in the main mash. This safeguards the mashing operation and provides an
improved wort, which reaches the correct and higher side of the desired
Fig. 4.4-1 and 4.4-2 show the activity of Termamyl BrewQ and Termamyl SC DS
Even so, there are still brewers who maintain that with good malt and standard
comparison:
37 C
60 C
90 C
300
Termamyl
225
150
BAN
75
10
10
pH
Fig. 4.4-1. Influence of pH on the activity of Novozymes Termamyl BrewQ at different temperatures.
(Activity curves for the conventional alpha-amylase Novozymes BAN shown for comparison)
Substrate: 0.5% soluble starch Stabilizer: 30-60 ppm calcium
<85-88 C>
100
100
80
60
Peak activity
Peak activity
80
40
20
Activity
60
High acid
Increasing temperature
High alkalinity
Increasing pH
58
59
Sorghum
When using sorghum as an adjunct, higher dosages of Termamyl are
Rice and maize (corn) grits and purified starch from maize (corn) and cassava
Rice and maize (corn) are widely used adjuncts that require separate
DS, as it works at a much lower Ca2+ level, below 20 ppm, and offers better
liquefaction. From laboratory liquefaction trials see Fig.4.5-1 and Table 4.5-1
100
Viscosity (cP)
80
60
1800
1400
1000
600
200
A, B, C
15
A
B
C
30
45
D
E
60
15
30
45
60
Nitrogen
Time (minutes)
nitrogen (FAN) yeast nutrients may be the result. This can be counteracted
resulting viscosities
by using a protease, such as Neutrase 0.8L /1.6 L, in the malt mash in order
to extract more nitrogenous compounds from the malt. This topic will be
discussed further in the relevant chapter of Fermentation control with FAN
0.025
0.05
0.025
0.025
0.05
Ca(OH)2 concentration*
100
100
100
Inactivation
* % of adjunct
6.1
6.1
6.5
7.3
6.5
Ultraflo Max, Neutrase 0.8 L/1.6 L, Termamyl Classic, Termamyl BrewQ and
optimization.
Table 4.5-1.
Based on this and other experiences, our recommendations for rice and maize
(corn) liquefaction are as follows as an initial trial:
Follow the liquefaction profile for maize (corn) with a minimum 15-30
minutes in the temperature range 85-95C
A Termamyl SC DS dosage of 0.2 kg/ton adjunct depending on the mill
setting/grits size and liquefaction regime
pH 5-6
A water-to-adjunct ratio of 2.2:1 4:1
For purified starch made of corn or cassava it is recommended to use the
same process as for grits.
For the various forms of cassava (e.g. pellets and cake) please contact Technical
Services at Norvozymes as the solution is dependent on the process conditions
and equipment in use and on the starch quality delivered.
60
61
Thermostable -amylase
Hydrolyzes 1,4--glucosidic linkages in amylose and amylopectin. Gelatinized starch is rapidly broken
down into soluble dextrins and oligosaccharides.
Declared activity
120 KNU_T/g
3.2.1.1
Physical form
Brown liquid
Production method
Thermostable -amylase
Hydrolyzes 1,4--glucosidic linkages in amylose and amylopectin. Gelatinized starch is rapidly broken
down into soluble dextrins and oligosaccharides.
Declared activity
240 KNU_T/g
3.2.1.1
Physical form
Brown liquid
Production method
Thermostable -amylase
Hydrolyzes 1,4--glucosidic linkages in amylose and amylopectin. Gelatinized starch is rapidly broken
down into soluble dextrins and oligosaccharides.
Declared activity
3.2.1.1
Physical form
Brown liquid
Production method
62
63
Chapter 5.
65
Efficiency and time in wort separation and beer filtration are key brewing
The optimal working conditions for the Ultraflo series and Finizym 250 L are
Ultraflo is added to the mash-tun during mash-in, starting when ca. 1/3 of the
brews per day, and ensuring high volumes of beer per filter run.
Finizym 250 L is added to the fermentor at the start of fermentation. Although,
The filtration enzymes include currently Ultraflo Max, Ultraflo L, Ultraflo XL and
Finizym does not work at its optimal temperature the solution can still be
Finizym 250 L.
Key benefits
0.1 kg/ton when using well modified malt and 0.25 kg/ton when using
barley (< 14Plato)
0.15 kg/ton when using well modified malt and 0.3 kg/ton when using
barley (>14Plato)
If very short mash filtration time is requested, trials have shown that
higher doses can be effective
Possibility to use High Gravity Brewing and Very High Gravity Brewing
Secure minimal investment in brewhouse and beer filtration capacity
Higher extract yield
66
67
During the malting of barley, the cell walls are broken down, and most of the
-glucans are degraded to lower molecular weight, less viscous polysaccharides,
as seen in Fig. 5.2-2. Arabinoxylans are not broken down to the same degree
The efficiency of separating wort from the mash, and later on the efficiency
beer. The malt derived from -glucanases and xylanases are not very heat
stable as they are. They are inactivated at temperatures above 50C and will
as can be seen in Fig. 5.2-1. They are also present in other cereal grains, but in
different amounts and ratios. Barley, oats and sorghum have more than twice
as much -glucan compared with xylans, while it is the opposite with wheat
in the wort and beer. The lower the modification of the malt, the higher the
and rye. Maize (corn) and rice have only limited amounts of these compounds,
amount of solubilized high molecular weight -glucans and xylans, giving rise
so their contribution to filtration issues is not a factor. Please see the raw
to inefficient and long lasting wort separation, and rapid pressure build-up
Ferulic acid
Acetic acid
Arabinose
Arabinoxylan
-glucan
Arabinoxylan
Days
Protein
Carboxypeptidase
Arabinoxylan
-glucan
Arabinoxylan
Protein
-glucan
Arabinoxylan
Arabinoxylan
Ferulic acid
Fig. 5.2-2.
A. Barley grains malted for 6 days showing sprout and acrospire development and half cut kernels
Cell wall model
Bamforth et al., J. Inst. Brew., 4. 2001, 235239.
Cell wall 1
Middle lamella
Cell wall 2
stained with the fluorescent dye Calcofluor making the cell wall degradation visible.
B. Close up of Calcofluor stained thin section of barley grain showing cell wall degradation in
detail. Intact cell walls show light blue fluorescence. Degraded cell walls have no fluorescence.
-glucans and arabinoxylans are very hydroscopic; they absorb water readily.
Wheat, rye and sorghum are cereal grains that are also regularly malted. For
They create high wort viscosity, reducing mash filtration speed dramatically.
wheat and rye malt, the modification pattern is similar to that of barley malt,
These components also become rather greasy when they absorb water, so
where the arabinoxylans and -glucans are broken down to minor and less
they stick to other grain components and to filter aids and filter membranes.
-glucans and arabinoxylans can also stick to starch molecules, making them
68
69
All enzymes in the Novozymes Ultraflo series contain both -glucanases and
xylanases, but of different types. Only Ultraflo Max contains the GH-10 family
as seen in Fig. 5.3-1 and arabinoxylans , as seen in Fig. 5.3-2 to low viscosity
polysaccharides. The enzymes are more heat stable than malt enzymes. The
enzymes will only be inactivated at 70-75C, so they will stay active during
the entire mashing, resulting in improved wort separation and beer filtration
Glucanase only
Ultrao L
Ultrao XL
Ultrao Max
Viscosity (cP)
Glucose
Cellobiohydrolase
-1,3-1,4-1,4-1,4-1,4-
2.2
2.1
2.0
1.9
0
-glucosidase
Endo--1,3(4)-glucanase
CH2OH
H
H
O
O
OH
H H
OH
OH
H
CH2OH
H H
O
H
O
OH
O
H
CH2OH
H H
50
75
100
125
150
175
200
225
250
275
300
Dosage (ppm)
H
OH
CH2OH
OH
CH2OH
OH
25
Endo-glucanase, cellulase
OH
Fig. 5.3-3. Lowest viscosity level delivered by Novozymes Ultraflo Max at all dosage levels
OH
Fig. 5.3-1. Structure of mixed-linked 1,3-1,4 -glucans, also showing the action points of various
High Gravity Brewing, and Very High Gravity Brewing, with efficient mash
glucanases
Arabinose
Ferulic acid
Endo-xylanase (GH10)
HOH2C
H
OH
HO
H
H
H
H
HO
1
O
O
H
HOH2C
OH
H
OH
H
H
O
O
O
HO
HO
HOH2C
1
H
OH
4
H
OH
H
H
OH
HO
20
21
22
23
24
25
26
27
28
1
H
Plato
O
H
19
H
1
O
1
H
H
3
O
HO
4.8
4.6
4.4
4.2
4.0
3.8
3.6
3.4
3.2
3.0
2.8
2.6
2.4
2.2
2.0
Ultrao Max
Glucoronic acid
Viscosity (mPa*s)
Endo-xylanase (GH11)
Acetyl
Ultrao L
CH2OH
Fig.5.3-4 Wort viscosity as function of gravity when no enzymes, Novozymes Ultraflo L and Ultraflo
Max are added
Fig. 5.3-2. Structure of arabinoxylans, showing the action points of various endo-xylanases. Two
types of endo-xylanases are present in filtration enzymes: the GH-10 family (Glucoside Hydrolase)
can cut the xylose backbone into the right chain lengths for improved filtration better than the
GH-11 family, resulting in lower viscosity of wort and beer
70
71
two enzymes is less pronounced, but for beer filtration Ultraflo Max is always
superior. This can be seen in Fig. 5.3-5.
Fig. 5.4-1 5.4-3 show the influence of temperature and pH on Ultraflo Max,
Ultraflo L and Ultraflo XL performance under brewing conditions. Fig. 5.4-4
Ultrao Max
analytical conditions.
4
0
200
400
600
800
1000
100
1200
Fig. 5.3-5. Volume beer filtered as function of differential pressure showing significant improved
beer filtration when using Novozymes Ultraflo Max compared with standard filtration enzyme
Performance (%)
80
60
40
20
0
55
60
65
The low wort and beer viscosity results in significantly slower differential
70
75
Temperature (C)
pressure increase across the filter over time, resulting in more filtered beer per
filter run and less beer loss. Compared with no enzyme use, up to 50% longer
beer filtration cycles can be achieved, and compared with filtration enzymes
having the family GH-11 xylanase, 25-30% more beer through the filter can
been achieved.
-glucan-reducing activity
The effective breakdown of the cell walls accomplished by the Ultraflo enzymes
raw material quality.
100
Performance (%)
allows for higher extract yield in the order of 0.5 to 2.0% depending on the
Extract
Viscosity-reducing activity
80
60
40
20
0
4.0
4.3
4.6
4.9
5.2
5.5
5.8
6.1
pH
72
73
performance of Ultraflo L.
100
80
80
Performance (%)
Performance (%)
100
60
40
20
60
40
20
0
0
55
60
65
70
55
75
60
65
-glucan-reducing activity
Viscosity-reducing activity
80
Viscosity-reducing activity
-amylase activity
100
80
Performance (%)
Performance (%)
100
60
40
20
60
40
20
0
0
4.0
4.3
4.6
4.9
5.2
pH
74
75
Temperature (C)
Temperature (C)
-glucan-reducing activity
70
5.5
5.8
6.1
4.0
4.3
4.6
4.9
5.2
5.5
5.8
6.1
pH
75
Fig. 5.4-4 A and B show the influence of temperature and pH on the activity of
Finizym 250 L.
Use of exogenous enzymes for the reduction of wort and beer viscosity is the
most widespread enzyme application in the brewing industry, and it is one of
the first to have been regularly used throughout the years. The first filtration
enzymes only contained -glucanase activity, but today most filtration enzymes
100
80
contain both -glucanase and xylanase activities. The most advanced enzymes
60
have the xylanase of the GH-10 family, which secures the lowest wort and beer
40
viscosity.
20
Filtration enzymes are often added to all brews to level out fluctuations in
0
20
30
40
50
60
70
Temperature (C)
All Novozymes filtration enzymes break down the unmodified cell walls from
barley malt or from unmalted barley. The more intact the cell wall materials,
the higher the dosage of enzymes required to attain acceptable brewhouse
performance and beer filtration. The most advanced filtration enzymes, like
Ultraflo Max, provide significantly better performance, especially for beer
100
80
60
Choice of enzyme
40
The correct enzyme solution should always fulfill the needs of the brewer.
20
Evaluation of cost versus benefit is very important! If capacity and time is the
0
2
brewers focus, there will be a need for higher gravity, shorter mash separation
time, and longer beer filtration cycles. In this case the lowest possible viscosity
pH
is highly desirable, and Ultraflo Max is the answer. Ultraflo Max is well suited
Fig. 5.4-4 B. pH dependency of Novozymes Finizym 250 L
for well modified malt, moderately modified malt, and blends of barley and
well modified malt, up to 25% barley.
If gravity is relatively low (< 14 Plato), and the number of beer filtration cycles
is not critical, Ultraflo L or Ultraflo XL can fulfill the brewers needs. The choice
between Ultraflo L and Ultraflo XL is related to the quality of the malt and the
grist. Ultraflo XL is a more robust enzyme that can deal with moderately modified
malt, inhomogeneous malt, and barley and malt blends up to 2530 % barley.
Ultraflo L is more suited for use with well-modified malt, which is demonstrated
in Fig. 5.5-1.
76
77
Viscosity
Glucan
mg/L
Filtrate
cP
Plato
ml
13.6
120
13.5
2.00
100
13.4
450
1.95
80
13.3
300
1.90
60
13.2
150
1.85
40
13.1
where -glucan, wort viscosity, extract yield and filtration are measured.
1.80
20
13.0
900
2.10
140
750
2.05
600
Ultrao L
Ultrao XL
Ultrao L
Ultrao XL
Viscosity
Glucan
mg/L
Filtrate
mPa *s
Plato
ml/20 min
Fig.5.5-1. Laboratory mashing test showing the higher efficiency of Novozymes Ultraflo XL on
under modified malt versus Ultraflo L, due to the broader range of activities in the former.
For non-barley cereals and their respective malts containing significant levels of
-glucan and xylan, Novozymes filtration enzymes can also be employed for
wort and beer filtration improvements.
2.90
60
16.0
750
2.75
50
15.9
600
2.60
40
15.7
450
2.45
30
15.6
300
2.30
20
15.4
150
2.15
10
15.3
900
2.00
00
15.1
A A B
B C C D D
100 150 75 125 50 100 100 200
A A B
B C C D D
100 150 75 125 50 100 100 200
For wheat and rye with arabinoxylans as the main cell wall components,
Ultraflo Max is absolutely the preferred enzyme. In the case of sorghum, both
Fig. 5.5-2. Simple laboratory test showing the difference in performance for 4 different filtration
enzymes. A: Novozymes Ultraflo L, B: Ultraflo XL, C: Ultraflo Max; D: Standard -glucanase with
-amylase side activity
For an industrial scale evaluation it makes sense to test the various enzyme
This product is typically used when the brewer knows in advance the presence
solutions over a period of time, for example,1-3 months, and collect data such as:
of un-filtered wort with high -glucan levels that will give rise to problems in
filtration and may manifest as haze in the packaged beer. Some brewers prefer
cellar. When using difficult raw materials, this has shown to be valuable for
78
79
Two types of filtration enzymes, A: -glucanase and GH-10 family xylanase and
modified malt per brew. Dosages were 1.8 kg of Ultraflo Max versus 2.5 kg of
B: -glucanase and GH-11 family xylanase, were tested against no enzyme use,
standard -glucanase per brew. The average trial data are summarized in the
control, for a period of 3 weeks in a brewery using well modified malt and maize
tables 5.6-2 and 5.6-3 below. Analyses of the first worts showed low -glucan
(corn) grits. High gravity brewing was performed with first wort at > 20Plato,
values for both wort types, as expected. However, Ultraflo Max treated
and final wort at 17Plato. The dosage was 0.15 kg/ton of both enzymes.
wort was significantly lower in arabinoxylans than the wort treated with the
traditional -glucan product, as seen in Table 5.6-2.
Mash filtration was significantly improved for both enzymes versus control
Arabino-xylan
(ppm)
-glucan
(mg/l)
25,4
1045
< 15
25,8
145
< 15
Enzyme
7% higher flow
Beer filtration only improved when using enzyme A:
0.2 bar/2000 hl beer lower pressure versus B and control.
Filtration enzyme with
GH-10 family xylanase
No
enzyme
76
76
75
59
62
65
170
165
160
0.65
0.85
0.85
Table 5.6-2.
Table 5.6-1. Brew house performance and beer filtration improvements by use of exogenous
filtration enzymes
Extract yield
0.5%
Brewhouse capacity
Table 5.6-3.
80
81
Declared enzyme
Declared enzyme
-glucanase (endo-1,3(4)-)
Declared activity
250 FBG/g
Declared activity
700 EGU/g
250 FXU-S/g
Physical form
Liquid
Physical form
Liquid
Production method
Production method
Novozymes Ultraflo L
Declared enzyme
-glucanase (endo-1,3(4)-)
Declared activity
45 FBG/g
3.2.1.6
Side activities
Physical form
Liquid
Production method
Novozymes Ultraflo XL
Declared enzyme
-glucanase (endo-1,3(4)-)
Declared activity
45 BGU/g
3.2.1.6
Side activities
Physical form
Liquid
Production method
82
83
Chapter 6.
Attenuation Control
and Light Beer Production
84
85
Globally, one of the fastest growing beer styles in recent years has been the
Preferably, all attenuation enzymes should be added to the mash tun at mash-
in. Alternatively, these enzymes can either be added to a separate process tank
prior to the kettle or into fermentation. Please note that when fermentation
packaging to ensure that no residual enzyme activity exists in the beer, or that
dextrin material. The result is a highly attenuated beer. A beer made this way
no substrate is left in the final beer. Dosage is calculated based on total grist
will have 25-30% fewer calories than a normally attenuated beer, assuming the
proper fermentation.
Malt worts produced under standard brewing conditions with traditional raw
Novozymes offers a broad range of attenuation control products to allow
apparent degree of fermentation (ADF) of 80-85%. Both RDF and ADF are
used to describe the degree of attenuation of the wort (the latter (ADF)
does not take into account the lower density of alcohol compared to water
in the final gravity of the fermented beer). Attenuation is a measure of the
degree to which sugars (i.e. glucose, fructose, maltose, maltotriose) in the
Key benefits
qualities
Produce super-attenuated malt base for flavored alcoholic beverage
production
Increasing the attenuation level by 4-5% utilizing the same amount of raw
materials
OH
CH2OH
H
HO
OH HO
HO
O
HO
OH
OH
CH2OH
O
O
OH
OH
Amylopectin
OH
OH
OH
300 - 600
Amylose
O
HO
CH2OH
OH
O
HO
O
HO
O
HO
HO
86
87
DP 6
DP 4
DP 3
DP 2
Amyloglucosidase (glucoamylase)
Glucoamylases are typically the first choice for a brewer to produce highly
1. Enzymatic breakdown
Glucoamylase
Maltogenic -amylase
and glucoamylase
are efficient enzymes that produce a strong effect on wort attenuation at even
relatively low dosages.
Pullulanase
Glucoamylase
-amylase on amylopectin.
Oligosaccharide
chains
Glucose
Maltogenic -amylase
Glucose and
maltose
Pullulanase and
maltogenic -amylase
-amylase
-amylases cleave -1,4-glucosidic linkages in starch, as do glucoamylases,
and maltose from amylose and maltose, glucose, and limit-dextrin from
Glucose and
maltose
Amylopectin
-amylase
but act upon random locations on the starch molecule. They yield maltotriose
Glucose
Maltose
Long chain
Breakdown of starch:
1. Glucoamylase attacks the -1,4 and 1,6 links from the non-reducing end to produce glucose
2. -amylase attacks -1,4 links to produce malto-oligosaccharides of varying lenght
3. Maltogenic -amylase attacks the second -1,4 links of a oligosaccharide from the non-reducing end to produce maltose
4. Pullulanase attacks -1,6 links to produce un-branched chains. The pullulanase enzyme normally need an -amylase or
maltogenic -amylase pre-treatment before this enzyme is active to producing maltose.
Fig. 6.3-3. Amylopectin breakdown by glucoamylase, -amylase and pullulanase to glucose and
maltose
88
89
100
100
80
60
40
20
0
80
60
40
20
0
4,0
4,5
5,0
5,5,
6,0
45
50
55
60
pH
65
70
75
80
85
70
75
80
85
Temperature (C)
look at the activity curves for each based on temperature and pH. Select an
enzyme solution that has significant activity and stability where you want to use
it in either mashing or fermentation.
attenuation enzymes. It is clear that from a pH point of view, all enzymes have
significant activity in the typical pH ranges encountered during brewing. From a
temperature perspective, Attenuzyme Core (and Attenuzyme Pro) and
100
Fig. 6.4-1 illustrates the temperature and pH activity curves for Novozymes'
100
80
60
40
20
0
60
40
20
0
4,0
AMG 300L BrewQ have high activity between 60C and 70C and would be
80
4,5
5,0
5,5,
6,0
45
50
55
60
pH
more suitable for mashing application than Fungamyl BrewQ, which undergoes
65
Temperature (C)
Fungamyl BrewQ
100
100
80
60
40
20
0
80
60
40
20
0
3,0
4,0
5,0
6,0
7,0
8,0
10
20
pH
30
40
50
60
70
Temperature (C)
Novozym 26062
100
100
80
60
40
20
80
60
40
20
0
0
2,0
3,0
4,0
5,0
pH
6,0
7,0
30
40
50
60
70
Temperature (C)
90
91
enzyme selected or making sure that no more substrate is left in the final beer.
RDF
ADF
Typically heat is used to inactivate (or denature) the enzyme after its activity is
70-75
85-90
75-80
For use in the brewhouse, wort boiling will completely eliminate any remaining
enzymatic activity that may be present. For use in fermentation, typical
pasteurization (tunnel or flash) conditions will inactivate only Fungamyl BrewQ.
Limited activity will remain from Novozym 26062, but significant activity
80-90
90-95
95-100
Option
Enzymes
Dosage Range
Units
(per ton grist or hL beer)
Point of addition
Fungamyl BrewQ
0.5 to 5.0
g/hL
Start of fermentation
1.2 to 3.5
kg/ton
Mashing-in
+ Novozym 26062
kg/ton
0.35 to 1.0
kg/ton
Mashing-in
2.4 to 3.6
0.25 to 0.75
kg/ton
Mashing-in
1.2 to 2.4
kg/ton
0.15 to 0.5
kg/ton
Mashing-in
4.0 to 8.0
g/hL
Start of fermentation
1.2 to 3.6
kg/ton
Mashing-in
2 to 5
g/hL
Start of fermentation
12 to 20
g/hL
Attenuzyme Core
Attenuzyme Core
+ Novozym 26062
Attenuzyme Pro
Fungamyl BrewQ
+ Novozym 26062
Fungamyl BrewQ
+ Novozym 26062
will remain from Attenuzyme Core, AttenuzymePro and AMG 300 L BrewQ
6.0 to 18
kg/ton
+ Novozym 26062
6.0 to 18
kg/ton
Attenuzyme Core
2.0 to 6
kg/ton
Attenuzyme Core
1.5 to 5
kg/ton
pasteurization.
+ Novozym 26062
2.4 to 4.8
kg/ton
0.25 to 5.0
kg/ton
Attenuzyme Pro
It can be seen from Table 6.5-1 that the most efficient methods for producing a
Novozym 26062 with addition to the mash tun at mashing-in. Fig.6.5-1 below
mashing into the mash tun can smooth out small variations in attenuation.
illustrates the broad range of attenuation targets that can be reached with
Table 6.5-1 outlines recommended starting points for enzyme dosages to alter
enzyme dosage.
Attenuzyme Pro 1.85 kg/ton
Attenuzyme Pro 0.95 kg/ton
Performance %
100
95
90
85
80
75
0
30
60
90
120
180
240
Minutes at 64 C
92
93
that lautering and/or mash filtration times are as short as possible with good
Description
Declared enzyme
Hydrolyzes (1, 4)- and (1, 6)--D-glucosidic linkages at the non-reducing ends of polysaccharides to
produce glucose.
needed. This will manifest in less free amino nitrogen (FAN) in the wort. The
Declared activity
300 AGU/mL
levels of FAN should be measured in the wort, and if on the low side, should
3.2.1.3
Physical form
Liquid
Production method
performance.
The increased attenuation and increased amount of alcohol formed should be
taken into account when calculating the amount of raw materials used. For
a given strength of alcohol in the beer, lower amounts of raw materials are
should be at ca. 15-18 mg/L/P. If FAN is low, use of Neutrase 0.8 L BrewQ or
Neutrase 1.6 L during mashing can be beneficial for fermentation performance.
Which attenuation solution is best for me?
When choosing an attenuation solution, there are different decision factors a
brewer can consider to select the most appropriate product.
Declared enzyme
Declared activity
1600 AGU/g
ease, including shorter mashing times, lower enzyme dosages and the ability to
3.2.1.3
Physical form
Liquid
Production method
50%, increasing brewhouse capacity while saving time and energy. If the
brewer wishes to address attenuation adjustment in fermentation, the best
solution is Fungamyl BrewQ and possibly Novozym 26062.
94
95
Glucoamylase that hydrolyzes (1, 4)- and (1, 6)--D-glucosidic linkages at the non-reducing ends
of polysaccharides to produce glucose. Pullulanase that hydrolyzes (1,6)--D-glucosidic linkages in
pullulan, amylopectin and glycogen to produce smaller fragments of linear dextrin.
Declared activity
Physical form
Liquid
Production method
A heat-stable pullulanase which accelerates production of highly fermentable worts when used in
conjunction with a glucoamylase.
Declared enzyme
Pullulanase
Declared activity
400 PUN/g
3.2.1.41
Physical form
Liquid
Production method
A classic fungal -amylase used for increased starch breakdown, facilitating higher alcohol output.
Declared enzyme
-amylase
Declared activity
800 FAU-F/g
3.2.1.1
Physical form
Liquid
Production method
96
97
Chapter 7.
Fermentation control
with FAN optimization
98
99
The FAN recommendation for all-malt wort is 180 to 220 mg/L (at 12 P) or
Free Amino Nitrogen (FAN) for growth, which translates into acceptable and
adjunct (e.g. barley, corn, sorghum or rice) are employed, low FAN levels in the
resultant wort can occur.
For FAN increase, Novozymes offers brewers Neutrase 0.8 L BrewQ and
Neutrase Xtra 1.6 L.
Neutrase products provide consistent, higher levels of FAN, when the brewer
requires it, based on malt modification and choice of raw materials. These
Key benefits
The optimal working conditions for Neutrase are 45-55C and pH 5.5-7.5. It is
internal peptide bonds. With normal malt, no more than 30-40% of the
Recommended dosages for high adjunct ratios or under modified malt for FAN
generation:
Amino acid
Endo-protease
N-terminus
Exo-protease
Figure 7.3-1. Protein structure and the effect of endo and exo-proteases
100
101
Fig. 7.4-1 7.4-3 show the influence of temperature and pH on Neutrase activity
under analytical conditions without the stabilizing effect of proteinaceous substrates.
Increase of FAN (%)
35
100
80
60
30
25
20
15
10
5
0
40
0.2
0.4
0.6
0.8
1.2
20
0
30
40
50
60
70
Fig. 7.5-1. %-Increase of Free Amino Nitrogen by Novozymes Neutrase 0.8 L BrewQ addition
Temperature (C)
Example 1:
Brewing with adjuncts in a decoction process of 60% malt and 40% rice
Liquefaction with Termamyl BrewQ and FAN adjustment with Neutrase in main
mash during protein rest.
100
80
The result show an increase of around 13-20% in FAN level with the addition
60
of 0.4 kg/ton of Neutrase BrewQ, and 20-26% with the addition of 0.8 kg/ton
40
20
0
4
40
40
40
40
40
40
14 (CC) +
46 (MT)
14 (CC) +
46 (MT)
14 (CC) +
46 (MT)
60 (MT)
60 (MT)
60 (MT)
0.25
0.25
0.25
Neutrase BrewQ
(kg/ton of malt)
0.40
0.80
0.40
0.80
90
100
135
190
180
180
Extract (P)
12.0
12.1
12.2
12.3
12.3
12.3
pH
Rice (%)
133
151
154
156
188
197
Malt (%)*
Fig. 7.4-2. Influence of pH on the activity of Novozymes Neutrase at 45C
65 C
60 C
55 C
45 C
25 C
100
80
60
40
20
0
10
20
30
40
50
60
Time (minutes)
102
103
Example 2:
Brewing with 60% malt and 40% barley
Main mash regime: 52C/30 64C/35 67C/15 73C/10 78C/05
Cereal cooker regime: 55C/15 75C/10 85C
The result show an increase of around 30% in FAN level with the addition of
Declared enzyme
Neutral proteinase
Declared activity
0.8 AU_NH/g
3.4.24.28
Physical form
Liquid
Production method
Trial 1
Trial 2
Trial 3
Trial 4
0.60
0.60
0.60
0.60
Trial 1
Trial 2
Trial 3
Trial 4
0.30
0.30
0.30
0.20
0.20
0.20
Neutral proteinase
0.20
0.20
Declared activity
1.6 AU_NH/g
0.40
0.40
3.4.24.28
Analytics (16P)
Trial 1
Trial 2
Trial 3
Trial 4
Physical form
Liquid
FAN (mg/l)
149
196
151
195
Production method
-glucan (mg/l)
52
51
53
103
1.950
1.937
1.938
1.985
Viscosity (mPa*s)
104
105
Chapter 8.
Diacetyl control
106
107
one of the most offensive off-flavors in Pilsner-type beer, based on the taste
threshold 0.02 to 0.15 mg/L depending on beer style, brand and taster.
Key benefits
and reduced into the much less flavor-active compounds acetoin (3-hydroxy-2butanone) and 3-hydroxy-2-pentanone. This can be seen in Fig. 8.2-1.
No Diacetyl off-flavor
Shorten, or even by-pass rate-limiting warm maturation (diacetyl rest)
L-Threonine
Pyruvate
Glucose
Glycolysis
-acetohydroxy -acetolactate
-butyrate
Glyceraldehyde
Isoleucine
Leucine
-3-phosphate
+
O
O
Valine
Dihydroxyacetone
-phosphate
2,3-Pentanedione Diacetyl
O
OH
3-Hydroxy
-2-pentanone
OH
O
Acetoin
The working conditions for Maturex 2000 L are 10-45C and pH 4.0-7.0.
Fig. 8.2-1. Formation and reduction of diacetyl and 2,3-pentanedione during yeast fermentation of
Maturex 2000 L is dosed into the cold wort in the fermenting cellar at the
wort.
the results are not only pH and temperature dependent, but also related
extra time needed for avoiding diacetyl off-flavor, from weeks to 2-5 days.
Depending on the adjunct ratio, wort concentration (Plato), yeast type, and
physical environment, the rate of diacetyl reduction is variable in time and
temperature requirements and not easily predicted. Therefore, the time needed
to reduce diacetyl to an acceptable level below the flavor threshold can vary
significantly.
108
109
mg/L
1.4
12
1.2
10
1.0
0.8
0.6
0.4
0.2
14
P/ Reference
P/ Maturex treated
DA/ Reference
DA/ Maturex treated
2,3-P/ Reference
Days
-aceto-lactate to diacetyl. But this reaction is slow when compared with the
action of Maturex 2000 L transforming the precursor directly to acetoin, so at
Fig. 8.3-2. Comparison of diacetyl and 2,3-pentandione formation and removal in fermenting wort
P/ Reference P/Maturex treated DA/Reference DA/ Maturex treated 2,3 P/Reference 2,3-P/Maturex treated Flavor threshold for diacetyl
O
CH3
CH3 O
Spontaneous
oxidative decarboxylation
O-
Slow reaction
CH3
Diacetyl
OH
precursor to these compounds, and only when they are excreted from the yeast
cells and present in the fermenting beer. This is demonstrated in Fig.8.3-3.
Yeast reductase
-aceto-lactate
O
Fast reaction
CH3
H
C
Sugar
CH3
CH3
Pyruvate
OH
Maturex 2000 L
COOH
Acetoin
CH3
CH2
CH3
CH
NH2
COOH
CH3
CH3
H
C
Maturex 2000 L
CH3
Fast
reaction
CH3
O
C
C
OH
wort, but taken up again by the yeast, so the diacetyl level is under the flavor
threshold at the end of fermentation.
O
O
-acetolactate
O
Slow
oxidation
CH3
CH3
Diacetyl
Extracellular -acetolactate
Minor amounts of diacetyl are still formed in the Maturex 2000 L treated
C
OH
CH3
OH
Acetoin
Valine
Diffusion
The formation of diacetyl does not need to be completely suppressed, but
CH3
CH3
H-
CH3
Diacetyl
Diffusion
CH3
CH3
OH
Acetoin
Fig. 8.3-3. Generation and reduction of diacetyl within the yeast cell and in the extracellular
medium in the presence of Novozymes Maturex 2000 L
110
111
100
80
60
40
20
0
10
20
30
40
50
60
Temperature (C)
100
80
60
40
20
0
3
pH
112
113
Maturex 2000 L is a unique enzyme specially designed for the brewing industry,
Maturex has been used for many years for quality and cost saving reasons. It
Fig. 8.6-1 and 8.6-2. In this case, the diacetyl rest was reduced from 4 to 2 days
is used year round or during special periods with tight capacity, for example,
Extract (% P)
be used, if the yeast produces extra diacetyl as a result of stress. This could be
Diacetyl
Temperature (C)
Temperature
Extract
15
14
13
12
11
10
9
8
7
6
5
4
3
2
1
0
-1
-2
0.85
0.80
0.75
0.70
0.65
0.60
0.55
0.50
0.45
0.40
0.35
0.30
0.25
0.20
0.15
0.10
0.05
0.00
4.20
4.18
9 10 11 12
4.18
4.15
Diacetyl(mg/L)
13 14 15 16 17 18 19 20
4.16 4.20
pH
Time (days)
Standard measurements for VDK and diacetyl, for example, ANALYTICA EBC
9.24.1 and 9.24.2 can be used to evaluate the effect of Maturex 2000 L.
Both methods can be used to measure the actual amount of VDK or diacetyl, as
well as the total VDK and diacetyl potential.
Extract (% P)
Diacetyl
Temperature
Extract
15
14
13
12
11
10
9
8
7
6
5
4
3
2
1
0
-1
-2
0.85
0.80
0.75
0.70
0.65
0.60
0.55
0.50
0.45
0.40
0.35
0.30
0.25
0.20
0.15
0.10
0.05
0.00
To measure the total VDK and diacetyl potential, the wort or beer must be
heat treated prior to analysis. Heat treatment at 60C for 90 minutes converts
the precursor -acetolactate and -acetohydroxy-butyrate to diacetyl and
2,3-Pentandione, respectively.
Temperature (C)
5
4.20
4.17 4.20
8
4.20
9 10 11 12
4.20
Diacetyl(mg/L)
13 14 15 16 17 18 19 20
4.15
pH
Time (days)
Please note that Maturex 2000 L works on the precursor released into the
fermenting wort. These precursor can be excreted by yeast, and also by some
Fig. 8.6-2. Trial with Novozymes Maturex 2000 L addition (2g/hl) and reduced diacetyl rest
114
115
Cleaning
to Filter
Lagering -1C
Cooling
Maturing 7 C
0.07 mg/l was reached when final attenuation was reached. This was after 84
Tank
20C. Using Maturex 2000 L dosed at 1g/hl, the level of acceptable diacetyl of
7
1
14
21
1,4
Temperature (C)
20
1,2
18
16
1,0
14
0,8
12
10
Extract (%)
0,6
0,4
6
4
Diacetyl(mg/l)
Days
22
0,2
2
0
0,0
24
48
72
96
120
144
Declared enzyme
2000 ADU/g
Declared activity
Fig. 8.6-3. The effect of Novozymes Maturex 2000 L on the diacetyl content
4.1.1.5
Physical form
Liquid
Production method
to Filter
Lagering -1C
Cooling
Maturing 7 C
1
2
Tank
3
4
5
6
7
8
1
0
14
21
Days
116
117
References
References
1
S. Home
Cellulases: a novel solution to some malting and brewing problems
EBC Congress, 1983
Wolfgang Kunze
Technology Brewing and Malting
International Edition, VLB Berlin, 1996
N.H. Aschengreen
Brewing Technology
Internal brewing compendium: 1998, update Novozymes 2003
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