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OUTLOOK HAZY OR BRIGHT?

Lance Shaner

Owner

Omega Yeast Labs

Yeast Strains to Dial in Haze

Florida Craft Brewers Conference 2024

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Who are we?

Omega Yeast labs

Chicago, IL / St. Louis, MO

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High quality, pitch-ready liquid yeast. Handful of microbiologists, homebrewers, professional brewers and craft beer fans who made it our express purpose to make brewing easier and better for everyone.

Lance Shaner

Co-owner/Founder

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What is haze?

    • Haze = Turbidity
    • The cloudiness of a fluid cause by large numbers of individual particles that are generally invisible to the naked eye
    • Analogous to fog or smoke in air

Maplewood Brewery & Distillery. Chicago, IL

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Where does haze come from?

MALT

protein

polyphenol

B-glucan

YEAST

cell wall

glycogen

HOPS

phenolic compounds

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Types of haze

Chill Haze:

    • Non-covalent interactions
      • hydrogen and hydrophobic bonding
    • Reversible
      • Occurs at 0°C but goes away at 20°C

Permanent Haze:

    • Covalent bonding
    • Often occurring with increasing age, repeated cooling and warming, pasteurization, oxidation

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Malt and hops

Proteins

    • Major malt-derived proteins
      • Proline-rich (different from foam stabilizing proteins)
      • Barley Hordein (20% proline)
      • Wheat Gliadins (15% proline)
      • Haze generating smaller <40kDa peptide/proteins

Polyphenols

    • Proanthocyanidins
      • Monomers, dimers, trimers and higher polymers of catechin, epicatechin and gallocatechin

Kahle EM, Sarnkow M and Jacob F. JASBC 2020.

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What kind of haze are we talking about?

NEIPA levels

Sample

Turbidity Measurement

Drinking water

0.05–1.5 NTU

Lager

5–20 NTU

Porter / Pale Ale

20–200 NTU

Hazy IPA

200–1000 NTU

Orange juice

300–900 NTU

Milk

>4000 NTU

NTUs are a measurement of haze

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Measuring Turbidity

Tubidimeters/nephelometers

In the lab:

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Anton Paar HazeQC ME

    • Measures 90º, 25º, and 0º

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Hach Turbidimeter

    • Measures 90º, and/or 0º

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Optek Haze Control DT9011

    • Measures 90º, 11º, and 0º

In the brewery:

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Haffmans OptHaze-i

    • Measures 90º, 25º, and 0º

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Separators

    • Measures 90º, and/or 0º

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Wort: All barley malt (2-row) for target 15°P

Strains: Haze Positive OYL-011, Haze Neutral OYL-004

Pitch Rate: 10 million/ml

Temperature: 70°F

Fermentation End Point: 14 days

Dry Hop Amount: 2 lb/bbl (8g/L)

Dry Hop Addition:

    • Control (no dry hop)
    • Knockout (in fermentor pre-pitch)
    • Day 1
    • Day 2
    • Day 3
    • Day 4
    • Day 7
    • DDH (½ Day 1, ½ Day 7)

Haze Assay

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    • Yeast Strain Specific:
      • “Haze-positive”
      • “Haze-neutral”
    • How does dry hopping play in?
      • Dry-hop timing
      • Hop variety
      • Hop products
      • Dose rates
    • HZY1 - a gene for haze

Talk Outline

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Dry hop timing and yeast choice

Haze Positive

OYL-011 British V

Haze Neutral

OYL-004 West Coast Ale I

Dry Hop Timing

Dry Hop Timing

Pictures at 14 days. From left to right: Control (no dry hop), Knockout, Day 1, Day 2, Day 3, Day 4, Day 7, DDH (Day 1 and 7)

This dramatically impacts degree of haze

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Dry hop timing and yeast choice

Haze independent of terminal plato

KO

Day 1

Day 2

Day 3

Day 4

Day 7

DDH

OYL-011

OYL-004

750

500

250

0

Haze (NTU)

100

OYL-011

OYL-004

2.5

2

1.5

1

0.5

0

Terminal Gravity (Plato)

KO

Day 1

Day 2

Day 3

Day 4

Day 7

DDH

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Grouping strains:

Haze Positive

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Hazy IPA strains/Hefe/Kolsch

    • OYL-011 British V (Juice/London III/Foggy London)
    • OYL-017 Kolsch I
    • OYL-032 East Coast Ale
    • OYL-061 Voss Kveik
    • OYL-043 Point Loma
    • OYL-015 Scottish Ale
    • OYL-005 Irish Ale
    • OYL-021 Hefeweizen
    • OYL-044 Kolsch II
    • OYL-006 British I

Haze Neutral

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American IPA strains

    • OYL-004 West Coast I (Chico)
    • OYL-052 DIPA Ale (Conan)
    • OYL-200 Tropical IPA

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Other Strains

    • OYL-030 Belgian Wit
    • OYL-024 Belgian Ale
    • OYL-071 Lutra Kveik
    • OYL-016 British Ale VIII (Fullers)
    • OYL-091 Hornindal Kveik
    • OYL-057 Hothead Kveik
    • OYL-114 Bayern Lager

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Haze (NTU) with 2 lb/bbl dry-hop addition at day 7

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    • Hopping Rate
    • Hop Variety
    • Hop Products
    • Early Dry Hoping vs. Late Dry Hopping?

How does dry hopping play in?

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Haze increases with dry hopping rate

0.5

1

2

4

750

500

250

0

Haze (NTU)

Dry Hopping Rate (lb/bbl)

KO

Day 4

Day 7

*fermented with OYL-011 haze-positive strain

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Haze is dependent on hop variety

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More haze variability between hop varieties and hop product types

Overall trends show variety specific impacts on haze

Cryo and T-90 pellets show different haze potential

Supplier 1

Supplier 2

Cascade

Centennial

Citra

Mosaic

400

300

200

100

0

Cascade

Ekuanot

Loral

Simcoe

Amarillo

Centennial

Mosaic

Sabro

T-90 (2 lb/bbl)

Cryo (1 lb/bbl)

400

300

200

100

0

Haze (NTU)

Haze (NTU)

*fermented with OYL-011 haze-positive strain

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Double dry hop experiments

Early hop additions limit haze potential

Cascade

Centennial

Citra

Mosaic

KO

Day 7

1/4 KO and Day 7

300

200

100

0

Haze (NTU)

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Hazy Control

Pictured Left

Experimental

Pictured Right

Knockout

-

0.5 lb/bbl

Day 4

2 lb/bbl

2 lb/bbl

Haze

487 NTUs

34 NTUs

ABV

7.60 %

7.61%

AE

2.93

2.86

pH

4.66

4.7

Pilot Brew

Can we make a bright NEIPA with an addition of a 0.5 lb/bbl of knockout hops?

Recipe Details

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Wort: 100% 2-row, 16.7ºP

Strain : Haze Positive OYL-011

Pitch Rate: 10 million/ml

Temperature: 70°F

Fermentation End Point: 14 days

Whirlpool Hops: 1 lb/bbl Citra, Wai-iti, and Mandarina Bavaria

Dry Hop:

    • Hazy Control: Day 4 alone
    • Experimental: Knockout + Day 4

Cold crashed at Day 14 for one week and transferred to keg

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Summary of initial work

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Haze

No Haze

Yeast

OYL-011 or haze-positive strains

OYL-004 or haze-neutral strains

Dry-hop timing

Mid-late

Early

Dry-hop rate

High

Low

Hop Product/Variety

Variable, more to learn!

…what makes a yeast haze-positive?

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HZY1 makes yeast haze-positive!

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How we found

Genetic backcrossing experiment analogy

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HZY1

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How we found

Following the genetics

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HZY1

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How we found

Following the genetics

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HZY1

Haze-positive isolate

Haze-positive isolate

Haze-neutral isolate

Haze-neutral isolate

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Large repeat expansions in HZY1 gene in haze-positive strains

Region of interest contains HZY1 gene

The HZY1 gene has large repeat expansions in OYL-011

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Large repeat expansions in HZY1 gene in haze-positive strains

HZY1 is expanded in haze-positive strains

HZY1 gene

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Large repeat expansions in HZY1 gene in haze-positive strains

Found new haze-positive yeast that had an expanded HZY1 gene

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Deleting HZY1 in Brewing Strains Reduces Haze

Delete HZY1

Reduce Haze

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What about sensory when the haze difference is obvious?

Does Hazy = Juicy???

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The differences were not obvious… more tasting required!

“Honestly a guess,” while another correct taster commented, ​“I’m having difficulty picking out similar beers.” Across the three panels, the theme of the comments was primarily that all four samples were incredibly similar.

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What does this all mean?

    • Making hazy and non-hazy beers can be calculated!
    • Know what yeast and hop combinations work best for the targeted style

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Style

Yeast

Dry Hop

Pilsner

Haze-neutral (lager strains)

Consider an early dry hop and keep addition rates low

WC IPA

Haze-neutral (West Coast I, Lutra, even a lager strain)

Consider an early dry hop and hop varieties with minimal haze

Hazy IPA

Haze-positive (British V, Voss, Point Loma)

Only late dry hop and hop varieties with maximum haze

Hefe

Haze-positive (OYL-021)

Small late dry hop to help with haze stability

* Email me if you have questions or are interested in trialing strains!

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Evidence for HZY1 changing in the brewery

    • Grimm Artisanal Ales noticed their house strain was becoming less and less hazy. They had been serial repitching for 300 batches.

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    • Their strain proved to have lost its haze positive phenotype in our flask assay.

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    • After sequencing the Grimm house strain, we observed it had lost the N-terminal repeat expansion.

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    • What drives the selection for this change in HZY1?

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How did brewing yeast evolve to be hazy?

    • Hzy1 is a yeast mannoprotein that is heavily glycosylated. The expansions could change the surface of the cell to promote it to be more “buoyant”

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      • Maybe… Top Cropping Selected for Buoyant (HZY1)

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      • Cone Harvesting Selected for Flocculant (FLO genes)

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    • Still more to learn about Hzy1, and the mechanisms for yeast-dependent haze in dry hopped beers!

Jamie Bogner skimming the open top fermentor at the RRBC pilot brewery

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THANKS!

Lance Shaner

Owner

Omega Yeast Labs

lance@omegayeast.com

Stay in Touch

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Haze occurs across a range of knockout pHs

Kncokout pH was adjusted with lactic acid and NaOH between pH 4.5 and 6 and fermentations were dry hopped on day 7

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Yeast blends to for stable haze

Control

KO

Day 4

Day 7

OYL-011

OYL-052

OYL-011:OYL-052 50:50

OYL-205

OYL-011:OYL-205 50:50

500

400

300

200

100

0