Practical Enzymatic Brewing An intermediate exploration of Brewing Enzymes
Presentation Summary This seminar is a companion to a previous presentation, Basic Enzymology for Brewing. This presentation is focused on: • A review of the sources and historic/typical roles of endogenous enzymes • An overview of exogenous enzymes available • How to most effectively use both types of enzymes in the brewing process
Alan B. Windhausen Head Brewer || Quality Trainer Holidaily Brewing || Brewers Association
Learning Objectives • Know the origins of both endogenous and exogenous enzymes • Be able to push endogenous enzymes and increase their effect • Have a greater understanding of sources and varieties of exogenous enzymes • Have an appreciation of the limitations of exogenous enzymes, and their potential downsides • Gain a sense of new products made possible by smart enzyme usage
Outline Endogenous Enzymes in Beer Process Optimization and Other (Review) Enzyme Uses • Origins, Malting • Mash and Lauter • Adjuncts Exogenous Enzymes • Kettle • Origins • Fermentation • Available Enzymes • Filtration
Endogenous Enzymes in Beer Review with focus on optimization – for full examination of process, please watch Basic Enzymology for Brewing
Malting Consists of: • Steeping • Hydrating the grain, starts the process of growth. • Germination • Endogenous enzymes break down stored nutrients • And kilning • Halts modification, creates flavor. Modification of barley, two paths Michael Lewis and Tom Young, Brewing (2 nd edition), 2002)
Germination 1 – The hydrated embryo eats sugars in its immediate vicinity. 2 – Gibberellins (hormones that start modification) are released from the Scutellum and 3 – Specific enzymes get released or produced to break down the endosperm. Enzyme production during malting Hans Sejr Olsen. Enzymes in brewing. Biokemisk Forening. 2008
Germination (cont’d) The types of enzymes produced are: Cellulose digesting And Starch Proteases : enzymes: digesting enzymes: (α- and β-amylase) – break Break down proteins, the Break down the walls that down starches into sugars. grain uses throughout the enclose the starch granules. Brewers need these enzymes process for various to be created but to not break purposes. Principal enzymes are β- down the starches yet. glucanases (for β-glucans), Creates Free Amino and pentosanases (such as Limit-dextrinase and other de- Nitrogen, critical for yeast xylanases). branching enzymes are also health. created. Check out the CBC 2020 presentation on Malt COA’s!
Germination (cont’d)
Germination (cont’d) Solubilization: - Xylanase - Acetyl xylan esterase - Feruloyl esterase - Arabinofuranosidase - Carboxypeptidase Hydrolysis (breakdown) - Endo- and Exoglucanases - Glucosidases - Xylosidase
Germination (cont’d) Protein matrix breakdown: (Proteases) - Endopeptidases - Exopeptidases
Germination (cont’d) Cell Wall breakdown during modification Courtesy of Canadian Malting Barley Technical Centre Barley modification by day Gianinetti, Theory in Biosciences, 2008
Germination (cont’d) Limit Dextrinase: • Present and active in mash for only short time: • An inhibitor is rapidly solubilized into the mash. Competitive inhibitor Kevin Ahern & Indira Rajagopal, Biochemistry Free & Easy, 2019 Adjuncts may increase the amount of limit dextrins (different starch ratios). Exogenously, pullulanases will serve this same function without inhibition. Non-Competitive inhibitor Kevin Ahern & Indira Rajagopal, Biochemistry Free & Easy, 2019
Kilning Goal is to suspend modification, leaving starches and amylases intact. Caramel and crystal malts are mini- mashes inside the kernel. • Sugars form, then are caramelized Exogenous enzymes that break down sugars may impact fermentability of malts / adjuncts! Munich 20L Malt vs Caramel 20L David Richter, Briess Malt and Ingredients – blogpost, Jan. 2018
Exogenous Enzymes Overview of origins and available enzymes
Exogenous Enzymes Enzymes added to the brewing process. Sources: Historically: • Bacteria or fungi. • α-amylase originally produced from • Yeast is common source. cattle and pigs. • Could be endogenous enzymes. • Barley and malt are still used (β- • Could also be GMO (tailor-made amylase, e.g.) enzymes). • Fruit still provides commercial • Heat-resistant fungi are popular. proteases. • Certain strains of E. coli are commonly used as well.
Exogenous Enzymes (cont’d) Partial list of enzymes available From Biokemi, BioZoom, issue 522
Exogenous Enzymes (cont’d) “There is a pervasive resistance of the brewers in North America to use enzymes. It is not the norm [unless] there is a special target not achievable by any other means.” “Enzymes are a sensible way to improve beer and to equalize differences from batch to batch. [However], when enzyme companies are saying, ‘Hey you can do things in a totally different way, you can use for example raw barley.’ Well you won’t get the same beer.” - Dr. Charlie Bamforth
Exogenous Enzymes Principal use of exogenous enzymes is Alternate use (and more common in US in addressing fluctuations in ingredients. craft) is novel product design. • Batch of malt has low FAN. • Brut IPA is prime example • Variability in filtration times due to • Low-cal, low carb products malt variation. • Extremely high / 100% adjunct • Diacetyl rests and concentrations mashes vary with yeast • Faster production times • Unusual mash regimes However, if used on known brands, triangle test!
Process Optimization and Other Enzyme Uses Mash, Adjuncts, and Lauter
Mashing Goals of Mashing: • Solubilize the 15-25% of malt matter that is readily soluble. • Gelatinize starches and convert into sugars usable by yeast in desired ratio for style and gravity. • Release and solubilize other desirable malt components (proteins, amino acids, yeast nutrients, etc.)
Mashing (cont’d) Enzyme Temperature Range Denatures pH Range Function α-amylase 150-160 o F (66-71 o C) ~170 o F (77 o C) 5.3-5.7 Cuts larger starches (Ca 2+ randomly stabilized) β-amylase 130-150 o F (54-66 o C) ~160 o F (71 o C) 5.0-5.5 Breaks down starch chains, linearly, into maltose Proteases 122-138 o F (50-59 o C) ~155 o F (68 o C) 4.6-5.3 Break down proteins 113-128 o F (45-53 o C) ~145 o F (63 o C) (increase FAN) (peptidase) β-glucanase 95-131 o F (35-55 o C) ~140 o F (60 o C) 4.5-5.5 Breaks down cell-wall materials et. al. Limit-dextrinase 95-140 o F (35-60 o C) ~150 o F (65 o C) 5.0-5.8 Breaks down sugars left behind by amylases, can be inhibited
Mashing (cont’d) From Biokemi, BioZoom, issue 522
Mashing (cont’d) Rough enzyme ranges in Mash John Palmer – How to Brew
Mash - Attenuation Control Starch-reducing enzymes courtesy of Dupont, annotated by author
Mash - Attenuation Control (cont’d) Enzyme activity on starches Kunze
Mash - Attenuation Control (cont’d) Brewer’s Window Jake McWhirter – Missionary Brewer Blog
Mash - Attenuation Control (cont’d) Amylase optimizations / uses: • Exogenous α-amylase to mash at β-amylase optimum (~142 o F, 62 o C) • Adding α-amylase and fungal α-amylase to low DP malt / increase fermentability • Adjunct additions to mash (~20% endo, up to 100% exo) • Reduce cost of running a cereal cooker / make kettle available as such (malt liquor, anyone?)
Mash - Attenuation Control (cont’d) Brew Process Courtesy of Yuengling & Son, Inc
Mash – Attenuation (cont’d) Amyloglucosidase (glucoamylase): • Creates glucose, not maltose. • Increases fermentability in standard mash & Real Degree of Fermentation • Can create lower-cal / lower-carb products Concern: • Stalled Fermentation • ‘Glucose suppression/repression’ “Giving yeast glucose before maltose is like giving a kid French fries – how on Starch-reducing enzymes earth will you get it to eat the Brussel courtesy of Dupont, annotated by author sprouts?” – Professor Michael Lewis
Mash – Attenuation (cont’d) Pullulanse (limit-dextrinase): • Cuts at branch points, reduces unfermentable dextrins • Increases fermentablility (RDF above 88% w/ other exo.) If used without other exo. enzymes: • Increase in RDF, fermentable ratios stay roughly the same! • No glucose suppression/repression! Starch-reducing enzymes courtesy of Dupont, annotated by author
Mash Filtration – Cellulases Enzyme Temperature Range Denatures pH Range Function Proteases 122-138 o F (50-59 o C) ~155 o F (68 o C) 4.6-5.3 Break down proteins 113-128 o F (45-53 o C) ~145 o F (63 o C) (increase FAN) (peptidase) β-glucanase 95-131 o F (35-55 o C) ~140 o F (60 o C) 4.5-5.5 Breaks down cell-wall materials et. al.
Mash Filtration – Cellulases (cont’d) Commercial enzymes might be marketed as: • β-glucanase (should be pure β-glucanase) • Xylanase (should be pure xylanase) • Pentosanases (often pure xylanase) • Cellulases (β-glucanase + xylanase, most likely) • Combination products (cellulases + protein/starch enzymes)
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