YEAST Wrangling The Many Flavors of Brewing Yeast CURT WITTENBERG FOR SOCIETY OF BARLEY ENGINEERS OCTOBER 4, 2017

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YEAST Wrangling The Many Flavors of Brewing Yeast CURT WITTENBERG FOR SOCIETY OF BARLEY ENGINEERS OCTOBER 4, 2017

Please distribute cups and beer Please keep beers in numerical order: 1-6. Please do not drink beers until you have been given instructions.

Fermentation Control The struggle to maintain a balance between competing reactions. Compound A Compound B Compound C

Fermentation is complex! Fructose Raffinose Starch Sulfur compounds Diacetyl Amino Acids + Esters Ketones Fusel Alcohols Glucose Acetaldehyde Ethanol + CO 2 Phenols Maltose Maltotriose Sucrose

Off flavors result from natural metabolism

A simplified view:

Multi-YEAST Tasting All beers are made with: Imperial Penny Blonde wort A White Labs yeast Some have one additional ingredient (see list) Evaluate the aroma and flavor. Attempt to identify all of the yeast flavors. Try to guess the yeast style and, if possible, the yeast strain. Beer 1 No additions Beer 2 No additions Beer 3 Dry hopped Beer 4 Dry hopped Beer 5 Dry hopped Beer 6 Table sugar

Off Flavors Associated with YEAST When good yeast go bad Acetaldehyde Acetic Acid Alcohol Diacetyl Estery/Fruity/Banana Fusel Alcohol Medicinal/Band Aid Phenolic/Spicy/Clove Sulfur compounds Sweet Yeasty/Soapy/Fatty

YEAST Management General rules for controlling off flavors produced by yeast. 1. Practice good sanitation Contaminating bacteria and yeast introduce many off-flavors 2. Use appropriate pitching rate Recommended to moderately high pitching rate gives lower off flavors. Long lag and growth give higher off flavors 3. Provide good nutrition, especially zinc Better nutrition = less stress = less off flavors 4. Use temperature control Cooler is generally better = less off flavors 5. Provide oxygen early; avoid oxygen late Oxygen supports growth but after growth phase leads to oxidation and ester formation 6. Don t cold crash too soon Yeast re-metabolize some off-flavor compounds once sugar is depleted More efficient at higher temperatures

Off-flavors re-utilized postfermentatio n Acetaldehyde Sensory Profile: green apple, green beer, latex paint, Jolly Rancher Present in all beers Appropriate in American Lagers Causes: An intermediate in alcohol production; can also result from oxygen when packaging. Prevention: Pitch healthy yeast, oxygenate early, allow time post-fementation at increased temperature for clean-up

Off-flavors re-utilized postfermentatio n Diacetyl (vicinal di-ketones) Sensory Profile: Buttery, butterscotch; slick mouth feel Present in all beers Appropriate at low levels in some styles, including English Bitters, Scotch Ales, Dry Stouts, and Czech Pilsner. Causes: Long lag phase, short post fermentation rest Prevention: Avoid by pitching healthy yeast, oxygenating early and include post-fermentation rest at >60F

Off Flavors associated with stressed YEAST Also appropriate properties of some beers Alcohol Sensory Profile: Alcohol (Ethanol) aroma and taste, can smell sweet and taste spicy or hot/solventy Present in all beers Appropriate in all beers Causes: excessive fermentables, light bodied beers mask alcohol flavors poorly Prevention: Avoid excessive fermentables in low body, low flavor beers Sweet Sensory Profile: Sweet, sweetness on finish that doesn t clear rapidly from palate Present in some beers Appropriate in some styles including Barleywines, Sweet Stouts, Tropical Stouts and others. Causes: Excessive unfermentable sugars, poor yeast health and nutrition. Premature removal of yeast Prevention: Pitch healthy yeast, good nutrition, oxygenating early and allow yeast to complete fermentation. Increase temperature late in fermentation. Pitch attenuative yeast

Off Flavors associated with stressed YEAST Appropriate properties of some beers Estery/Fruity/Banana (Isoamyl Acetate) Sensory Profile: Banana, pear, apple, berry, nail polish when high concentrations Present in all beers Appropriate in some styles including many British & Belgian beers and German wheat beers. Causes: Long lag phase, high temperature, low O2 early, good yeast nutrition; inappropriate yeast strain for style Prevention: Pitch sufficient healthy yeast, oxygenate early and control temperature. Pick appropriate yeast strain for style. Characteristic of some yeast strains

Off Flavors more common in specific YEAST strains Also appropriate properties of some beers Phenolic/BandAid/Medicinal/Smoky/Spicy/Clove (Eugenol; 4-vinyl guiacol) Sensory Profile: Large range of aromas and flavors (see above) Appropriate at low levels in some styles, including many Belgian beers and German wheat beers. Causes: Characteristic of some yeast strains, increased when stressed; wild yeast or Brett contamination; poor wort processing can also contribute Prevention: Pick appropriate strain for style; control temperature; watch sanitation

Off Flavors associated with stressed YEAST Fusel Alcohol Sensory Profile: Alcoholic; hot; bitter; solventy Inappropriate above flavor threshold, especially in lighter beers Causes: Insufficient pitch, high temperature; excessive time on yeast; contamination with wild yeast Prevention: Pitch sufficient healthy yeast and control temperature; remove beer from yeast when fermentation is completed Acetic Acid Sensory Profile: Slight to strong sharp vinegar flavor Appropriate at low levels in some styles including berlinerwiesse, wild and sour ales, some dry stouts. Causes: Some yeast strains, wild yeast, Brett, bacteria Prevention: Choose appropriate yeast strain, watch sanitation

Off Flavors associated with stressed YEAST Appropriate properties of some beers Sulfur compounds Sensory Profile: Range of off flavors including burnt match, rotten eggs, rotten vegetable, sulfury, cooked vegetables Present in all beers Appropriate at very low levels in some styles, especially lagers. Causes: Characteristic of lager yeast, especially when stressed by low oxygen, poor nutrition or low pitch rate; yeast autolysis; bacterial contamination Prevention: Pitch sufficient healthy yeast and provide good conditions; watch sanitation; remove beer from yeast following reasonable postfermenations rest. Characteristic of some yeast strains

Off Flavors associated with stressed YEAST Yeasty/Soapy/Fatty Sensory Profile: waxy, soapy, fatty or greasy sensation Inappropriate Causes: Yeast autolysis from severe yeast stress or death Prevention: Avoid by pitching healthy yeast and removing beer from yeast before excessive period post-fermenation; avoid severe yeast stress

Let s talk about the Big Brew Beers? Beer 1 Off -Flavors? Yeast Strain? Beer 2 Off -Flavors? Yeast Strain? Beer 3 Off -Flavors? Yeast Strain? Beer 4 Off -Flavors? Yeast Strain? Beer 5 Off -Flavors? Yeast Strain? Beer 6 Off -Flavors? Yeast Strain?

Insert Picture(s) The Reveal

The Reveal: The flavors of YEAST Identities of the Big Brew Beers Beer 1 WLP001 - California Ale Yeast Temperature: 68F Additions: None Brewer: Scott Woodward Beer 2 WLP007 Dry English Ale Yeast Temperature: 68F Additions: None Brewer: Scott Woodward Beer 3 WLP007 Dry English Ale Yeast Temperature: 68F Additions: Dry hop with 2 oz Neomexicanus Hops Brewer: Andy Gamelin Beer 4 WLP029 German Ale/Kölsch Yeast Temperature: 65F, then raise to 70F Additions: Dry hop with 2 oz Eldorado Hops Brewers: Michael and Lauren Gaggioli Beer 5 WLP500 Trappist/Monastery Ale Yeast Temperature: 68F Additions: dry hop with 2 oz Neomexicanus Hops Brewer: Andy Gamelin Beer 6 WLP545 Belgian Strong Ale Yeast Temperature: 68F then rise to 74 Additions: 1 lb Sucrose as Simple Syrup Brewer: Curt Wittenberg

Brülosophy YEAST xbeermnts Not so fast. Experiments revealing little or no detectable or undesirable effect: Temperature Pitching Rate Starter vs Direct Pitch Viability vs Vitality Starter Repitch vs Fresh Yeast Old vs Fresh Repitch

The origins of brewing yeast?

YEAST Genomics A little knowledge is a dangerous thing. The Relationship Between Brewing Yeast and Other Industrial Yeasts Gallone et al, Cell, 2016 (Miraglia, White Labs, Verstrepen Lab, Belgium) Gonçalves et al, Current Biology, 2016 (Sampaio Lab, Portugal) Sequence the genomic DNA of >150 industrial yeast strains Compare all based upon DNA sequence to each other and previously sequenced genomes of lager yeast, wild yeast and others. Construct unrooted cladistic tree based upon sequence data Establish the occurrence of mutations in genes known to be involved in domestication-related functions (i.e. sexual reproduction, off-flavor production, etc.)

Gallone et al, Cell, 2016 Two independent clades of ale yeast British, American, Belgian and German fall into the Beer 1 clade. Others fall into a smaller Beer 2 clade and a few into wine and mixed origin yeast. Ale yeast are very diverse even within the same clade indicating rapid evolution, probably due to defined growth conditions over many generations.

American Ale California Ale Wheat Kölsch Gonçalves et al, Current Biology, 2016 Ale yeast are mostly clustered into a group containing British, German and Wheat beer strains. Many ale yeast are more closely related to other industrial yeast and likely originated independently. Kölsch and British ale strains have mutations in genes required for synthesis of 4-vinyl guaiacol (clove phenolic) whereas most Wheat beer strains do not. Genes that regulate water movement in and out of cells are inactivated in almost all wine, beer and sake strains which helps cells grow in high sugar environments.

Gonçalves et al, Current Biology, 2016 Lager yeast placement in the cladistic tree is based upon the sequence of S. cerevisiae DNA sequences, ignoring S. eubayanus sequences. Lager yeast occupy a place between Wheat Beer and English-Irish Ale yeast, but more distant from German Alt-Kolsch yeast. There are two cladistic trees of Lager yeast, Frohberg and Saaz, suggesting two independent origination events, consistent with different S. cerevisiae and S. eubayanus chromosome structures.

Occurrence of Domestication-Related Characteristics in Industrial Yeasts Maltotriose Utilization Gallone et al, Current Opinions in Biotechnology, 20

Occurrence of Domestication-Related Characteristics in Industrial Yeasts 4-Vinyl Guaiacol Production

Occurrence of Domestication-Related Characteristics in Industrial Yeasts Sexual Reproduction

YEAST Improvement Using knowledge to direct the development of new yeast strains. Knowledge of DNA sequence and chromosome structure can inform us about requirements for specific growth and flavor characteristics. New yeast strains can be developed based upon knowledge of genes rather than guessing based upon sensory or other traits. Strain development can still use classical genetics without implementing genetic modification approaches, thereby, avoiding the GMO dilemma.

Thank you! Questions?