1. Practical Brewing Enzymology
Tobin L. Eppard
Coors Brewing Company
Rocky Mountain District - MBAA
May 11, 2005
MBAA – Rocky Mountain District

2. MBAA – Rocky Mountain District
What is in (and out) the box for today
A Brief Primer on Enzymes
Malting processes
Mashing processes
Practical Brewing Implications
Q&A Session
MBAA – Rocky Mountain District

3. 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
Enzyme/Substrate
Product(s)
MBAA – Rocky Mountain District

4. MBAA – Rocky Mountain District
What are Enzymes?
Protein
Structure is specific to function
How is shape determined?
Functional groups
Amino acids….
MBAA – Rocky Mountain District

5. MBAA – Rocky Mountain District
What are Enzymes?
Primary Structure
Secondary Structure
Tertiary Structure
Quaternary Structure
MBAA – Rocky Mountain District

6. MBAA – Rocky Mountain District
What do they do?
“Spontaneous” Chemical Reactions
Energy Barriers, EA - Energy of Activation
Reduction of EA (G) by as much as 1015
Energy Level B2 B1 A
EA of Enzyme Catalyzed Reaction C
MBAA – Rocky Mountain District

7. Enzyme Biochemistry Lock and Key Model Product(s)
Substrate
Enzyme/Substrate Complex
Enzyme
Enzyme
MBAA – Rocky Mountain District

8. MBAA – Rocky Mountain District
Enzyme Biochemistry
Enzyme “Process” in Biologic Sense
NEED Energy
RNA
Inhibition
AMINO ACIDS
DNA
Ribosome
Yeast
4 o Sequence
A-T-G-A-C-G
Functional Protein (Enzyme?)
3 o Sequence
2 o Sequence
1 o Sequence
MBAA – Rocky Mountain District

9. MBAA – Rocky Mountain District
Enzyme Biochemistry
Factors Influencing Enzyme Rates:
Endo vs Exo enzymes pH
Temperature
Products/Inhibition
Kinetics
MBAA – Rocky Mountain District

10. MBAA – Rocky Mountain District
Enzymes in Malting
What’s going on in barley?
Thousands of enzymes…
Cellulases
Proteases
Amylases
MBAA – Rocky Mountain District

11. MBAA – Rocky Mountain District
Enzymes in Malting
Barley Modification
Pictures: David A. Thomas, Coors Brewing Company and Kunze, Technology Brewing and Malting
MBAA – Rocky Mountain District

12. MBAA – Rocky Mountain District
Enzymes in Mashing
Mashing
Solubilize starches
and Amino Acids/peptides
Starches to reducing sugars
Denature protein
MBAA – Rocky Mountain District

13. MBAA – Rocky Mountain District

14. MBAA – Rocky Mountain District
Brewing Enzymes
Enzyme Action Opt. Temp pH
Amylase  67C 5.2
Amylase  62C 5.5
Proteases < 52C 5.5
 Glucanase – 52C 6.0
Glycosidic -1,4 Endoamylase
Glycosidic -1,4 (Non-reducing end C-4) Exoamylase
Solubilize Proteins and Polypeptides (FAN) Endo and Exoproteinases
Glycosidic β-1,4 and β-1,3 Gum/Cell walls, reduction in wort viscosity Endoglucanase
MBAA – Rocky Mountain District

15. *Exogenous Brewing Enzymes
Enzyme Action Opt. Temp pH
Amylase (HS) ~ 67C 5.2
Amyloglucosidase ~ 60C  5.5
(AMG)
Glucanase – 52C 6.0
Pullulanase
Glycosidic -1,4 Endoamylase
Glycosidic -1,4 Endoamylase
Glycosidic β-1,4 and 1,3 Gum/Cell walls, reduction in wort viscosity Endoglucanase
Glycosidic -1,6 (Random) Endoamylase
* - Not an exhaustive list, contact Novo or Genencor for more information
MBAA – Rocky Mountain District

16. MBAA – Rocky Mountain District
Practical Aspects of Brewing Enzymes…
MBAA – Rocky Mountain District

17. MBAA – Rocky Mountain District
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..
MBAA – Rocky Mountain District

18. MBAA – Rocky Mountain District
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
MBAA – Rocky Mountain District

19. MBAA – Rocky Mountain District
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 of 95%
MBAA – Rocky Mountain District

20. MBAA – Rocky Mountain District
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…
MBAA – Rocky Mountain District
Pitures: Bill Simpson (Cara Technologies)

21. MBAA – Rocky Mountain District
First Aide Kit
Beer Foam
Maximize solubilization of HMW Proteins
Consider raising your initial mash in temp (60-62C)
Hops +
Viscosity (higher PG)+
Gas +
Yeast Enzymes
Stress and high gravity – hydrophobic polypeptide reduction
MBAA – Rocky Mountain District

22. Quality System Most important of all…
Set Malt Quality parameters and hold supplier accountable to meet them
Extract - S/T or Kohlbach
Viscosity - Total Protein
Color - Friability DP
MBAA – Rocky Mountain District

23. MBAA – Rocky Mountain District
Final Notes on Exogenous Enzymes:
Safety Concerns (handling)
It’s a Soup of a larger cast of characters
Side Reactions
MBAA – Rocky Mountain District

24. 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)