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Home , Berliner Weisse, Brettanomyces, Flanders red ale, Sour beer

Souring Organisms

Brittney Berg Lallemand [email protected]

LALLEMAND BREWING Overview

LALLEMAND BREWING History and styles of sour

• styles have existed for centuries • What do we mean by Sour beer? • History and heritage of sour have been based in Europe, including: ➢

➢

➢

➢ Flanders Red

LALLEMAND BREWING Modern sours

• Growth in Belgian sour imports in 90’s influence US • Based on historic styles • Huge diversity in sour styles, flavors and creativity

LALLEMAND BREWING Souring applications and techniques

LALLEMAND BREWING Key microorganisms

• Bacteria ▪ producing bacteria - - • Non- Wild ▪ ▪ Lachancea • GMO souring yeast ▪ Sourvisiae

LALLEMAND BREWING Lactic Acid Producing bacteria

Pediococcus & Lactobacillus

LALLEMAND BREWING Pediococcus

• Lactic acid producing bacteria, can be <3.0 pH • Slow acting • Can grow in CO2 rich environments and in O2 environments • Found naturally on most fruits • Hop tolerant • A “ropey” diacetyl producer • Help from Brettanomyces!

LALLEMAND BREWING Lactobacillus

• Primary souring bacteria • Diverse range of sub species • Temperature sensitive • Softer and tangier lactic acid • Sugar Utilization

LALLEMAND BREWING Sources of LaB

• Laboratory (pure or mixed culture) • Bottle cultures • Nature • Yogurt • Un-mashed grain

LALLEMAND BREWING R&D Trials

LALLEMAND BREWING R&d trials: Fermentations (20C) (..F)

LALLEMAND BREWING R&D trials: Fermentations (30C)

LALLEMAND BREWING R&D trials: fermentations (40C)

LALLEMAND BREWING R&D trials: fermentations (50C)

LALLEMAND BREWING R&D trials: lactic acid production

LALLEMAND BREWING R&D trials: production

LALLEMAND BREWING R&D trials: Sensory characteristics

LALLEMAND BREWING Wild non-saccharomyces strains

Brettanomyces & Lachancea

LALLEMAND BREWING brettanomyces

• A wild yeast; large genetic diversity • Was commonly seen as spoilage yeast, gained popularity • Barnyard, horsey, band-aid & fruity esters • Tolerant to harsh conditions • Sugar utilization • Attenuation ▪ Good attenuators due to production of a range of alpha & beta- glucosidases • Secondary Metabolites ▪ Varies based on the composition of the , available oxygen, and time. ▪ Good at producing acetic acid even low levels of O2

LALLEMAND BREWING Common Brettanomyces strains

1. Brettanomyces bruxellensis • Most popular strain – widely used • Medium intensity Brett character • Commonly found in Belgian pales and • Typically starts as ‘horse blanket’ / leathery and become more light & earthy as time goes on 2. Brettanomyces lambicus • Most intense out of the bunch – very horsey, spicey/smokey flavor • Found in Lambics • Keep O2 low to avoid “nail polish’ – ethyl acetate 3. • Light intensity Brett, can take many months to fully develop noticeable characteristics • Often associated with fruity aromas – pineapple & mango

LALLEMAND BREWING Lachancea

• a lactic acid producing non-sachh yeast • Like most wild strains, it is a very slow fermenter • Can be used in primary or secondary ferms • Not as much know with this strain and not used as often in commercial • Known strains: L. fermentati and L. thermotolerans • New unknown Lachancea strain has been discovered and is believed to be genetically different from that of L.fermentati and L.thermotolerans •

LALLEMAND BREWING Lachancea fermentation data

Lachancea fermentation temperature 20°C; pitching rate 100g/hl 14.0 LYCC 8471, G120218 LYCC 8471, G120218, duplicate

12.0 LYCC 8467, 230118 LYCC 8467, 230118, duplicate

10.0

P) 8.0 °

6.0 Density Density (

4.0

2.0

0.0 0 24 48 72 96 120 144 168 192 216 240 264 Time (h)

LALLEMAND BREWING Lachancea lactic acid production

LALLEMAND BREWING Lachancea sugar metabolism

LALLEMAND BREWING New Technology

GMO Strain – Sourvisiae™ from Mascoma

LALLEMAND BREWING New technology: bioengineered strain

SourvisiaeTM is a bioengineered ale yeast (Saccharomyces cerevisiae) capable of producing lactic acid during primary fermentation: • Reproducible • Mono-culture • No other flavor compounds produced • Generally Regarded as Safe (GRAS)

LALLEMAND BREWING Sourvisiae™ Ethanol production

Ethanol 60

50 Parent Strain

40 Sourvisiae

30 g/L

Wild Yeast 20

10 Wild Yeast

0 0 2 4 6 8 10 12 14 Fermentation Time (days)

LALLEMAND BREWING Sourvisiae™ Lactic acid production

Lactic Acid 9

8 100% Wild type (WT) 7

6 100% Sourvisiae

5 g/L 4 Wild Yeast

3

Wild Yeast 2

1

0 0 2 4 6 8 10 12 14 Fermentation Time (days)

LALLEMAND BREWING Sourvisiae Pros/Cons

Pros • Will not hold up your kettle; saves time and money • So far no THP discovered through sensory Cons • GMO may not be for everyone – and that’s ok! • Difficulty predicting initial sourness, relying on blending in the end

LALLEMAND BREWING Final notes

• Know your final flavor/aroma wants • SOPs – stay clean! • Strive for continuous improvement and experimentation • If you’re going to brew sour beer, be prepared to dump it

LALLEMAND BREWING Questions?

Brittney Berg – Lallemand Brewing [email protected]

LALLEMAND BREWING LALLEMAND BREWING CBA Trials Control vs Sourvisiae

INGREDIENTS AND PROFILE

Malts 90% Great Western Two-Row, 10% Briess Caramel C60 Hops Bittering - Alchemy Aroma - Cascade Yeast Sourvisiae

Wort Analysis: Pitch rate: OG: 12.76 100g/bbl into 5 bbls of wort pH:5.52 Pitch Temperature: 65F BU:41 Primary Fermentation Setpoint: 66F SRM: 8.7 Aeration Type: Filtered Plant Air, ~12g/L over 10 minutes of cooling

LALLEMAND BREWING Trials - Sourvisiae CBA/Widmer Control vs Sourvisiae

Trial pH Comparison 6

5

4

3 Trial AE Comparison 2 14

1 12 10 0 0 24 48 72 Final 8 6 Sourvisiae pH Control pH 4

2

0 0 24 48 72 Final

Sourvisiae AE Control AE

LALLEMAND BREWING Introduction to Genetic Modification

Direct manipulation of genome with biotechnology: • Specifically tailored function

• Use of familiar, single template strain

• Yeast are great at accepting other ’ DNA

• Finer tuning than classical approaches

LALLEMAND BREWING