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Coors Brewery Institute
Practical Brewing Enzymology
Tobin L. Eppard
Coors Brewing Company
www.coors.com
Rocky Mountain District - MBAA
May 11, 2005
MBAA – Rocky Mountain District
Coors Brewery Institute
• What is in (and out) the box for
today
– A Brief Primer on Enzymes
– Malting processes
– Mashing processes
– Practical Brewing Implications
– Q&A Session
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Historically speaking..
• 1783 – Spallanzani noted a non cellular breakdown of meat by
gastric juices
• 1814 – Kirchoff, wheat paste converted starch to sugars…
• 1878 – Kuhne suggested Enzyme = “In Yeast”
• Catalytic Activity
Substrate
Enzyme
Product(s)
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Enzyme/Substrate
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What are Enzymes?
• Protein
• Structure is specific to function
• How is shape determined?
– Functional groups
– Amino acids….
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•
•
•
•
What are Enzymes?
Primary Structure
Secondary Structure
Tertiary Structure
Quaternary Structure
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What do they do?
• “Spontaneous” Chemical Reactions
– Energy Barriers, EA - Energy of Activation
• Reduction of EA (G) by as much as 1015
B2
B1
A
EA of Enzyme Catalyzed Reaction
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C
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Enzyme
Biochemistry
• Lock and Key Model
Product(s)
Substrate
Enzyme
Enzyme/Substrate
Complex
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Enzyme
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Enzyme
Biochemistry
• Enzyme “Process” in Biologic Sense
Inhibition
NEED
Energy
RNA
AMINO
ACIDS
DNA
Ribosome
Yeast
Functional
Protein
(Enzyme?)
4o
Sequence
3o
Sequence
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A-T-G-A-C-G
2o
Sequence
1o
Sequence
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Enzyme
Biochemistry
• Factors Influencing Enzyme Rates:
– Endo vs Exo enzymes
– pH
– Temperature
– Products/Inhibition
– Kinetics
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Enzymes in Malting
What’s going on in barley?
Thousands of enzymes…
Cellulases
Proteases
Amylases
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Enzymes in Malting
Barley Modification
Pictures: David A. Thomas, Coors Brewing Company and
Kunze, Technology Brewing and Malting
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Enzymes in Mashing
Mashing
• Solubilize starches
and Amino Acids/peptides
• Starches to reducing sugars
• Denature protein
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MBAA – Rocky Mountain District
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Brewing Enzymes
Enzyme
 Amylase
Action
Glycosidic -1,4
Endoamylase
Opt. Temp
pH
 67C
5.2
 Amylase
Glycosidic -1,4
(Non-reducing
end C-4)
Exoamylase
 62C
5.5
Proteases
Solubilize Proteins
and Polypeptides
(FAN) Endo and
Exoproteinases
< 52C
5.5
40 – 52C
6.0
 Glucanase
Glycosidic β-1,4
and β-1,3
Gum/Cell walls,
reduction in wort
viscosity
Endoglucanase
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*Exogenous
Brewing Enzymes
Enzyme
 Amylase (HS)
Action
Opt. Temp
pH
Glycosidic -1,4
Endoamylase
~ 67C
5.2
Amyloglucosidase
(AMG)
 Glucanase
Glycosidic -1,4
Endoamylase
~ 60C
 5.5
Glycosidic β-1,4
and 1,3 Gum/Cell
walls, reduction in
wort viscosity
Endoglucanase
40 – 52C
6.0
Pullulanase
Glycosidic -1,6
(Random)
Endoamylase
-
-
* - Not an exhaustive list, contact Novo or Genencor for more information
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Practical Aspects of Brewing
Enzymes…
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First Aide Kit
Extract Production
– Starch Gelatinization Temperatures
• Well Modified Malt(s)
– High DP malt OR
• Heat stable Alpha Amylase and Beta Amylase
– Adjunct usage rates
• Dilution of DP..
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First Aide Kit
Extract Separation
– Maximize Beta Glucanase (what little is
there)
• Consider, lowering your first mash-in
temperature
– Exogenous Beta Glucanase
– Minimize shear forces
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First Aide Kit
Wort Fermentability (more or less?)
– Exogenous Alpha Amylase, Beta Amylase
– Amyloglucosidase, Pullulanase
– Slightly lower conversion hold (64-65C)
– Slower ramp from conversion hold to mash
off (2ºC/min vs. 1ºC/min)
– Consider the use of kettle adjuncts like
HFCS… @DE of 95%
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First Aide Kit
Beer Filterability
–
–
–
–
Exogenous Beta Glucanase
Maximize Trub Formation (finings, boil)
Maximize cold maturation (if possible)
Add DE filtration first, before sheet filtration (if
possible)
– Reduce shear forces
– Yeast flocculation issues
– Secondary finings (isinglass) and
hydrogels/xerogels…
Pitures: Bill Simpson (Cara
Technologies)
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First Aide Kit
Beer Foam
– Maximize solubilization of HMW Proteins
• Consider raising your initial mash in temp (6062C)
– Hops +
– Viscosity (higher PG)+
– Gas +
– Yeast Enzymes
• Stress and high gravity – hydrophobic
polypeptide reduction
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Quality System
• Most important of all…
Set Malt Quality parameters and hold supplier
accountable to meet them
–
–
–
–
Extract
Viscosity
Color
DP
- S/T or Kohlbach
- Total Protein
- Friability
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Final Notes on Exogenous Enzymes:
– Safety Concerns (handling)
– It’s a Soup of a larger cast of
characters
• Side Reactions
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Discussion?
Pictures and Acknoledgements:
David A Thomas – Barley grain modification SEM(s)
Dr. Bill Simpson – CaraTechnologies, UK
Mathewson, Paul R. Enzymes – Practical Guides for the Food Industry Eagan Press Handbook Series, St. Paul
Minnesota, USA 1998 (sponsored by the American Society of Brewing Chemists)