100% Barley Brewing Made Possible
David Maradyn
Novozymes North America
Franklinton, NC, USA
MBAA-RMD Technical Summit
Fort Collins, CO
Friday June 25, 2010
Agenda
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Introduction and History
Ondea Pro Concept
Components of Ondea Pro
Barley Considerations
Amino Acid and FAN
Filtration
Milling
Lautering
Flavor
North American Experience
Conclusions Q&A
Introduction
 Brewing with increased amount barley is primarily driven
by the lower cost of barley compared with malt
 Other drivers includes
 Reduction of the CO2 emission
 Use of local raw materials
 Barley is usually added by mashing-in of a limited
percentage of barley, up to 30%
 The Novozymes Ondea® Pro concept enables for brewing
of 100% barley (or combinations between barley and
adjuncts)
 where the resulting beer can be blended with other brews
 or be sold as is
 This presentation is focused on the enzymatic solution
behind Ondea® Pro
History
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Replacement of malted barley with enzymes and raw
barley in the brewing process is not new …
Concept has been under investigation for over 50 years
In the early 1960’s researchers at Labatt Breweries of
Canada studied the feasibility of adding protolytic and
diastatic enzymes to raw cereal mash, such as
unmalted barley or corn, to produce a brewers wort
Claimed to produce an acceptable beer both analytically
and organoleptically
DENNIS. G.E . and QUITTENTON, R.C. U. S. Patent No. 3 ,081,172 (March 12, 1963)
History
 A subsequent publication revealed that the enzymatically
produced barley wort had issues compared to a
traditional malted barley wort
◦ Lower total nitrogen and fermentable sugars
 Other issues
◦ Difficulty in milling raw barley to produce a proper particle
size suitable for enzymatic hydrolysis
◦ Higher final diacetyl levels were found at end of fermentation
◦ Lower colloidal stability was found in the final product
 Conclusion that idea was still in development and needs
further research
LATTIMER. R.A.; LAKSHMINARAYANA, K.; QUIT-TENTON, R. and DENNIS, G.E. Inst Brew (Aust Section)
Proc 9th Conv. 1966, pp. 111-16.
Summary of the Ondea® Pro enzymatic solution
for brewing with 100% unmalted barley
 The attenuation is ensured
through a synergetic action
between
 Endogenous action of the barley
beta-amylase
 Exogenous added pullulanase and
alpha-amylase
 The fermentable amino acids are
provided by a synergistic action of
 Endogenous exo-peptidases
 An added protease
 Good lautering with a clear wort is
provided by a combination of well
adjusted mills and
 A filtration enzyme system
including both beta-glucanase
and xylanase components, and
 A lipase to ensure the wort clarity
Mashing Profile
Temperature
78-82 °C / 10 min
64 °C / 45-75 min
54 °C / 30 min
Time
 The synergies between the
enzymes are ensured through
 A three temperature step
infusion mashing profile
 With a pH of 5.6-5.8
The pullulanase is the key for the attenuation
performance of Ondea® Pro
 Starch is degraded by a synergetic
action of the added pullulanase and
alpha-amylase and the beta-amylase
from barley
Fermentable
sugars
Unique pullulanase Starch
added
glucose
 The main reason for the good
performance of the chosen
pullulanase is the temperature
profile (stability) which fits the
gelatinization temperature of
barley well.
Comparison of the temperature
activity curves at pH 5 of three
pullulanases
maltose
α-amylase
added
Barley
β-amylase
120
Comparison of the amount of unfermentable dextrin
(DP4+) of 3 pullulanases
100
Ondea Pro
Pullulanase
Pullulanase A
80
Pullulanase B
Dose
Ondea Pro
Pullulanase
Commercial
pullulanase A
Commercial
Pullulanase B
0
30.7
30.7
30.7
17.5 g EP/T
barley
17.1
20.1
27.5
Lab mashing trials. Ondea Pro is compared with 2
commercial pullulanases at equal dose in enzyme
protein. DP 4+ measured by HPLC
% of max activity
 The key is selection of the pullulanase
60
40
20
0
20
40
60
oC
80
The attenuation components provide a typical RDF but
with a very high maltose content
 Novozymes Ondea® Pro gives a
high maltose concentration
 A high RDF is ensured by a low
amount of dextrins
 Ondea ® Pro at 2 kg/t barley gives
around 70% RDF
 Maltose dominates the sugar
profile
Maltose conc in %
Sugar profile
Glucose
5.0 %
Fructose
1.5 %
Maltose
> 60 %
Maltotriose
< 15 %
Dextrins (DP4+)
< 20 %
Low glucose level leafs the
opportunity to add syrup and still
create a “maltose” profile
Results from lab scale mashing trials
Novozymes Ondea® Pro ensures processing also at
higher gravities; but it takes a dosage adjustment or
an extension of the 64°C saccharification rest
 High gravity reduces the level of
saccharification due to product
inhibition of the enzymes
 Ensuring the same level of
saccharification at higher gravities
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Fermentable sugars (DP-1,2 and 3) measured
after laboratory mashing trials with 2 kg Ondea®
Pro per t barley and increasing gravity
Either increase the Ondea® Pro dosage
(demonstrated below), or
Extend the 64°C saccharification rest
The sugar profile is kept constant by increasing
the amount of Ondea ® Pro.
The increase in enzyme dose is plotted against
gravity, defining the dose at gravity 1:5 as 100%
Different barley qualities requires different
enzymes dosages.
 Ondea® Pro has been tested on
close to 100 different barley
samples collected around the
world (as of April 2010)
 It works on approx 95% of the
tested varieties
 The few barley where it fails are
not suited for malting
 The observed variation in
generated FAN is well correlated
to the protein content
 We observe some variation in
needed enzyme dose to reach
the target attenuation
 The reason for this variation is
under investigation
 It is not correlated to the betaamylase content
 For each barley variety there is linear
correlation between the enzyme dose and
amount of non-fermentable dextrin’s
 Different barley varieties do therefore
appear as straight lines when then the
amount of non-fermentable dextrin’s is
plotted against the enzyme dose
Effect on DP4+ of different Ondea Pro
dosages applied at two barley varaities
% DP4+
25
24
23
22
21
20
19
18
17
16
15
Barley A
Barley B
1
1,2
1,4
1,6
1,8
2
kg Ondea Pro per t grist
Laboratory mashing. Different dosages of Ondea® Pro
was applied at standard 2 hours mashing with 45 min
saccharification rest. The unfermentable dextrins (DP
4+) were measured by HPLC
Synergy with barley peptidases provides
good yeast performance
 Novozymes Ondea® Pro’s proteolytic component works in synergy with the
endogenous enzymes from barley
 This is demonstrated by a comparison between the effect of Ondea® Pro on barley
with and without inactivation of the endogenous enzymes.
Barley wort needs
only
9 mg FAN/l/Plato
to secure a good
fermentation
The effect of the synergy with endogenous enzymes is demonstrated by a comparison
between a mashes at 50 °C with and without a pre-inactivation of the endogenous
proteolytic enzymes by a heat treatment at 75°C for 30 minutes
Barley wort produced with Novozymes Ondea® Pro has
significantly higher amount of fast absorption amino acids
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The amino acids are often divided
into 4 groups according to their
absorption by the yeast cell
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The amino acid profile from 100%
barley wort differs from malt wort
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The barley wort contains relatively
more of the easy fermentable
amino acids (Groups A and B) and
relatively less of the less
fermentable amino acids (Groups C
and D) - especially proline
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This explains the good
fermentability of the barley wort,
despite the lower FAN than found
in malt wort
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Having an amino acid profile more
suitable for the yeast leads to less
amino acid based Strecker
aldehydes - and in turn improved
flavor stability!
Relative amino acid compositions of malt and
barley worts according to the amino acid
fermentability groups
Average of 10 barley samples with the malt from the corresponding barley
Group A
Fast
absorption
Glutamic acid
Aspartic acid
Asparagine
Glutamine
Serine
Threonine
Lysine
Arginine
Group B
Intermediate
absorption
Valine
Methionine
Leucine
Isoleucine
Histidine
Group C
Slow
absorption
Glycine
Phenylalanine
Tyrosine
Tryptophan
Alanine
Group D
Little or no
absorption
Proline
FAN level and FAA profile of barley and malt wort in
pilot and industrial scale
Pilot trial
8 hl
FAN (12 %) mg/l
FAA Composition
Group A
Group B
Group C
Group D
Industrial trial
Barley
114
43.4%
27.0%
20.6%
9.0%
Malt
174
32.7%
21.7%
18.2%
27.3%
300 hl
FAN (12 %) mg/l
Barley
150
Malt
216
FAA Composition
Group A
Group B
Group C
Group D
38.8%
23.8%
23.6%
13.8%
30.1%
20.9%
21.1%
27.8%
Amino acid profile and FAN level of barley (mashed with 2 kg/t Ondea ® Pro) and malt wort in
pilot (8 hl) and industrial scale (300 hl).
• Industrial barley trials showed a little higher proline content
• 70% Group A and B FAA vs 54% Group A and B FAA Pilot barley vs malt worts
• 63% Group A and B FAA vs 51% Group A and B FAA Industrial barley vs malt
worts
100 % barley wort shows comparable
fermentation performance with less FAN
•
•
•
•
The FAN recommendations for malt brews are 120-220 mg/l (at 12 oP) or 10-18 mg/l/Plato
The FAN from 100% barley wort is lower; 108-170 ml/l (at 12 oP) or 9-14 mg/l/Plato
The barley worts have good fermentability
This leads to less unfermented FAN at the end of the fermentation
Lab fermentation of 12% malt wort and barley wort (made with Novozymes Ondea® Pro).
Fermentation temperature 12 °C, Yeast strain W34/70
100% Barley Beer have a better amino acid profile for yeast
growth that leads to less formation of Strecker aldehydes
(aroma components in wort and beer)
Amino Acid
Strecker Aldehydes
Flavor
Methionine
Methional
Potato
Phenylalanine
Phenylacetaldehyde
Sweet, green, floral
Leucine
3-Methylbutanal
Malty, burnt
Isoleucine
2-Methylbutanal
Malty
Wort aroma components from trials in µg/kg
Pilot Plant (8 hl)
Barley
Malt
Industrial Scale (300 hl)
Barley
Malt
3-Methylbutanal
31
115
99
230
2-Methylbutanal
11
51
38
110
Methional
13
12
19
93
Phenylacetaldehyde
56
142
85
174
109
325
245
607
Total Strecker aldehydes
The filtration and extract component ensures low
wort viscosity and high extract yields
 Increasing the barley inclusion in mashing leads to an increased viscosity
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This cannot be solved by traditional filtration enzymes
 The filtration system in Ondea® Pro keeps the viscosity low even for 100%
barley, though a combination of
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A beta-glucanase
A viscosity reducing xylanase
 The filtration enzyme also ensures a good extract yield
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Evaluated by low Extractable Extract in spent grain
The wort viscosity from lab scale mashing trials was measured at different
barley inclusion levels with and without addition of Novozymes Ondea® Pro
The turbidity reducing component ensures
low turbidity and facilitates lautering
The resulting free fatty acids from the lipase action in the mash are at the same
level as malt with an similar or lower amount of lipid oxidation product
 Barley wort (without use of the lipase in Ondea®
Pro) has very high turbidity and is very difficult to
lauter
 The lipase in Ondea® Pro reduces the turbidity of the
wort to well under the guideline of 80 NTU (20 EBC
unit)
Lipase added to laboratory mashing. The
turbidity was measured after filtration/lautering
 The lipase releases fatty acids similar to the level
found in malt based mashing
 Less lipid oxidation products are found in the barley
beer, likely because there is a lower level of fatty
acid in the beginning of mashing where the LOX is
active and oxygen are present
Laboratory mashing. The amount of free fatty
acids before lautering was measured by HPLC
Barley milling in combination with lauter tun
and mash filter
 Mashfilter in
combination with
hammer mill
 The fine grist of the
hammer mill is
preferable for the
enzyme system
 The wort separation
procedure needs no
changes. Filtration
speed and turbidity
showed the same or
improved
performances when
compared with all
malt brews.
 Lauter tun in combination with roller mill
 A barley kernel is less friable, and the endosperm is more
closely connected to the husk – compared to malt
 Milling barley with the same roller setup used for malt will
yield a much more coarse grist composition
 The finest possible grist composition is needed
 Six and four roller mills are preferable
 However, successful industrial trials have been
completed with two roller mills
 Optimized barley milling showed the following:
Optimized barley grist composition
Barley
Malt
Sieve 1
25 -30%
(18%)
Sieve 2
15-20%
(8%)
Sieve 3
33-40%
(33%)
Sieve 4
10-15%
(21%)
Sieve 5
2-5%
(10%)
Bottom
8-12%
(11%)
Flavor of 100% Barley Beer
 The concept has been tested in a number of pilot and production brews globally
 The majority of products produced were good-tasting beers – no defects
 The resulting sensory profiles have been different reflecting the brewmaster’s
optimization
 The barley beer taste can be optimized to resemble taste of a malt based beer
Flavour Characteristics
Mouthfeel - Overall Score
Intensity
5
Overall Score *
6
5
4
3
2
1
0
4
3
2
Malty/Grainy *
Aftertaste *
Sulphury
1
0
Fruity
Mouthfullness
Estery
Malt
Barley
Sweetness *
Bitterness
Malt
Sensory panel comparison of a malt and a similar barley based pilsner
Taste panel executed by: Centre for Malting and Brewing Science at
K.U.Leuven
Barley
North American Experience
 Industrial trials in North America have been limited to date
since product Ondea Pro launch in fall 2009
 One successful trial completed in US
 High-maltose/low-glucose wort obtained
 No lautering issues
 Low viscosity wort produced: similar beta-glucans, 80% lower
arabinoxylans than in comparable 100% malt wort
 Areas of attention
 Milling: further optimization should have been done, as the
grist we obtained was coarser than we desired
 Mashing: grist:water ratio, continuous stirring of the mash
is absolutely critical
 Lautering: bed is not as compact as when using 100% malt
– first wort ran off very quickly, no need to rake as deep
 Whirlpool: trub is not as compact as with 100% malt wort
North American Experience
100% unmalted barley beer prototypes
 Two sets of Ondea Pro beers have been produced in North
America in 2010 and presented at two brewing
conferences
 Craft Brewers Conference, Chicago IL, April 2010
 American Pale Ale
 ASBC MBAA Brewing Summit, Providence RI, June 2010
 Irish Stout
 IPA
 Helles
 All beers were very well received by conference
participants
 Ondea Pro is one more tool at the brewmasters disposal
for the creation of an interesting array of beers
Conclusions
 A enzyme solution – Ondea Pro - and process have been
developed to enable the brewing of great-tasting beer with
100% unmalted barley with existing brewing equipment
 Ondea Pro works in synergy with the endogenous enzymes from
barley
 The special pullulanase found in Ondea Pro is essential for the
attenuation performance
 Ondea Pro yields a wort with a relatively high maltose content
(> 60%)
 The amino acid composition of 100% unmalted barley wort
differs from malt based wort – higher group A and B amino acids
and lower proline content
 The lauter tun performance is ensured through the enzyme
system, milling adjustment and lauter tun management
 Flavor of 100% unmalted barley beer is similar to 100% malted
barley beer
THANK-YOU
for your
ATTENTION
Q&A