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MBAA – Rocky Mountain District
Coors Brewery Institute
Practical Brewing EnzymologyTobin L. Eppard
Coors Brewing Company
www.coors.com
Rocky Mountain District - MBAAMay 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
MBAA – Rocky Mountain District
Coors Brewery Institute 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
Product(s)
Enzyme/SubstrateEnzyme
MBAA – Rocky Mountain District
Coors Brewery Institute What are Enzymes?
• Protein
• Structure is specific to function
• How is shape determined?– Functional groups– Amino acids….
MBAA – Rocky Mountain District
Coors Brewery Institute What are Enzymes?
• Primary Structure
• Secondary Structure
• Tertiary Structure
• Quaternary Structure
MBAA – Rocky Mountain District
Coors Brewery Institute What do they do?
• “Spontaneous” Chemical Reactions– Energy Barriers, EA - Energy of Activation
• Reduction of EA (G) by as much as 1015
Ene
rgy
Lev
el
C
A
B1
B2
EA of Enzyme Catalyzed Reaction
MBAA – Rocky Mountain District
Coors Brewery Institute
Enzyme Biochemistry
• Lock and Key Model
Substrate
Enzyme/Substrate Complex
Product(s)
EnzymeEnzyme
MBAA – Rocky Mountain District
Coors Brewery Institute
Enzyme Biochemistry
• Enzyme “Process” in Biologic SenseNEED Energy
DNA
RNAAMINO ACIDS
Ribosome
1 o Sequence
2 o Sequence
3 o Sequence
4 o Sequence
Yeast
A-T-G-A-C-GFunctional Protein (Enzyme?)
Inhibition
MBAA – Rocky Mountain District
Coors Brewery Institute
Enzyme Biochemistry
• Factors Influencing Enzyme Rates:– Endo vs Exo enzymes– pH– Temperature– Products/Inhibition– Kinetics
MBAA – Rocky Mountain District
Coors Brewery Institute Enzymes in Malting
What’s going on in barley?
Thousands of enzymes…
Cellulases
Proteases
Amylases
MBAA – Rocky Mountain District
Coors Brewery Institute Enzymes in Malting
Barley Modification
Pictures: David A. Thomas, Coors Brewing Company and Kunze, Technology Brewing and Malting
MBAA – Rocky Mountain District
Coors Brewery Institute Enzymes in Mashing
Mashing• Solubilize starches
and Amino Acids/peptides• Starches to reducing sugars• Denature protein
MBAA – Rocky Mountain District
Coors Brewery Institute Brewing Enzymes
Enzyme Action Opt. Temp pH Amylase 67C 5.2
Amylase 62C 5.5
Proteases < 52C 5.5
Glucanase 40 – 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
Coors Brewery Institute
*Exogenous Brewing Enzymes
Enzyme Action Opt. Temp pH Amylase (HS) ~ 67C 5.2
Amyloglucosidase ~ 60C 5.5
(AMG) Glucanase 40 – 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
* - Not an exhaustive list, contact Novo or Genencor for more information
Glycosidic -1,6 (Random) Endoamylase
MBAA – Rocky Mountain District
Coors Brewery Institute 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
Coors Brewery Institute 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
Coors Brewery Institute 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%
MBAA – Rocky Mountain District
Coors Brewery Institute 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)
MBAA – Rocky Mountain District
Coors Brewery Institute 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
Coors Brewery Institute 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
Coors Brewery Institute
Final Notes on Exogenous Enzymes:– Safety Concerns (handling)– It’s a Soup of a larger cast of
characters• Side Reactions
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)