Escarpment Laboratories Inc.

Guelph, Ontario, Canada

© 2023 by Escarpment Laboratories Inc.

All rights reserved. No part of this document may be reproduced

or transmitted in any form without written consent from
Escarpment Laboratories Inc.
YEAST FLAVOUR
& BIOTRANSFORMATION

Your comprehensive guide to easy,

high-impact fermentation.
Table of Contents
About Escarpment Labs 5
Our Team 7
Yeast Flavour & Biotransformation Research 9
Yeast Aroma Compounds 17
Terpene Biotransformation 23
Thiol Biotransformation 27
Flavour Impact of Lactobacillus 31
Impact of Temperature on Yeast Flavours 33
Impact of Pitch Rate on Yeast Flavours 34
Beer Ideas 39
About the Guide 45
ABOUT US
We believe fermentation
should be easy.
How do we do it? We focus on our core
values:
Trust: We are focused on growing
Trust:
microbial cultures that our customers can
trust.
Knowledge: We help our customers
Knowledge:
become fermentation experts.
Community:
Community: We listen to our customers
and collaborate with them to solve
problems.

Escarpment Laboratories is a yeast producer

Escarpment Laboratories
based in Guelph, Ontario, Canada. We have
served the craft beer community in Canada,
the United States, and beyond for more than
8 years.

In that time, we have established ourselves as fermentation experts through

education, research, and tailored customer support. We were the first to publish
scientific research on Norwegian Kveik yeasts. We have developed many
innovative products for the craft beer industry including hybrid yeasts (Hydra,
Thiol Libre) as well as new products arising from applied research (Lactobacillus
Blend 2.0, Yeast Lightning Nutrient). Our new product development helps our
customers achieve new heights in fermentation.

This guide is a result of our efforts to share as much useful data as possible with
brewers to help make yeast and fermentation decisions easier, based on years
of applied scientific research.

5
 We focus on
knowledge, trust, &
community.
Our Team
We are brewers, scientists, and
fermentation nerds.

R&D
Our internal R&D team is focused on taking the needs of brewers and
translating them into new products and solutions to make fermentation easier.
Our R&D team not only develops new yeast strains but also new solutions
(Yeast Lightning Nutrient) and new processes to improve yeast production and
handling.

Experienced Brewers
Many of our staff are ex-brewers who bring their practical expertise to the
equation and help ensure we can offer the best possible products and support
for professional brewers. Our experienced staff support professional brewers
who are faced with a unique set of challenges every day.

Research Partnerships
We partner with universities and research institutes to accomplish our biggest
research projects. These partnerships give us access to equipment and
expertise that extends the value of the Escarpment Labs research ideas being
explored. Examples include investigating yeast flavour with the University of
Guelph, yeast mutations during repitching with the University of Washington,
and new yeast development with VTT Finland.

Education
We also believe in supporting the next generation of scientists and brewers.
We employ university co-operative education students and train them in key
brewing industry methods and offer them opportunity to contribute to the
brewing science knowledge base. We also offer frequent 1-day brewing
microbiology courses.

8
YEAST FLAVOUR AND
BIOTRANSFORMATION
RESEARCH

We help you using science,

not guess work.

Why does Escarpment Labs conduct yeast flavour research?

Brewers want to know exactly what flavours they can expect from a new yeast
strain. They also want to know what to expect if the operational conditions
change in the brewery (such as temperature and pitch rate). We have focused
for years on developing the resources to be able to answer these questions
properly.

What tools are used?

We use a diverse set of tools in order to help brewers optimize their house
yeasts and select new strains. Through our research partnerships, we have
access to sophisticated analytical tools that give us a clear picture of the flavour
potential in each yeast strain.

For yeast flavour metabolite analysis, we have used various analytical methods
including Gas Chromatography Mass Spectroscopy (GC-MS with various sub-
methods), High-Performance Liquid Chromatography (HPLC), as well as one of
the greatest analytical tools of all: the human nose (a.k.a. sensory analysis). We
use analytical and sensory data to help brewers choose a strain and optimize its
flavours.

Why does yeast make flavour?

There are still a lot of open questions here! Because we also want to know why
yeast makes different flavours, we are committed to combining analytical data
with genomic data to understand the genetics underlying our favourite yeast
flavour profiles. We have sequenced the whole genomes of all of our public
yeast strains.

We are now linking yeast genetics to their flavour output to understand how
they work. This will help us develop the next generation of beer yeasts and has
already informed the creation of new products such as Saison Maison, Hydra,
KRISPY, and Thiol Libre.
9
 Brewers want
to know exactly
what flavours
they can expect
from a new
yeast strain.
KEY YEAST AND
FERMENTATION
FLAVOURS

Alcohols
Yeast is capable of making a diverse range of alcohols which contribute to its
flavour. At low levels, these alcohols can contribute to flavour complexity. For
example, 1-octanol is perceived as citrus-like. However, at high levels, fusel
alcohols such as isobutanol and isoamyl alcohol can be perceived as “boozy”.
Different yeast strains also produce different amounts of glycerol, which
contributes to body and mouthfeel in beer.

Esters
Esters are perceived as fruity and floral flavours. There is a wide range of esters
that yeast is capable of producing, making this family one of the most interesting
to brewers! Broadly speaking, we can divide esters into fatty acid esters such as
ethyl hexanoate (pineapple-like) and ethyl octanoate (apple or cognac-like) and
acetate esters such as isoamyl acetate (banana-like) and phenylethyl acetate
(rose-like).

Phenols
Grain contains phenolic acids, which the grain plant produces as a stress-
protectant against fungi in the field. These flavourless phenolic acids can be
converted by yeast into volatile phenols such as 4-vinylguaiacol, which gives
Belgian beers their distinctive peppery-clove note. In many beers this note is
considered an off-flavour, leading to its common term of “POF” (phenolic off-
flavour).

Acids
Yeast is capable of making different types of acids, and some yeasts reduce the
pH of wort more than others. In typical yeasts, this acid production is minimal
and includes succinic acid and acetic acid. Specialized acid producing yeasts
(such as Lactic Magic) as well as souring bacteria are capable of producing a
wider range of acids, predominantly lactic and acetic acids but also citric acid.
These contribute to the tart and refreshing taste of sour beers.

11
Terpenes
Yeast is capable of both unlocking and transforming terpene compounds
(monoterpene alcohols) from hops. This can make a beer taste more citrusy,
fruity, or dank. Keep reading to learn more about terpene biotransformation.

Thiols
Certain yeasts are capable of unlocking thiol aromas from flavourless hop and
malt precursors. This can make a beer taste more tropical. Keep reading to learn
more about thiol biotransformation.

12
YEAST AROMA COMPOUNDS

LEGEND

For cleaner flavour profiles to let other ingredients shine through For high intensity yeast character

Note:
Aroma compounds are measured using HS-SPME-GC-MS. Concentrations are scaled to the compound’s sensory
threshold in beer to best show the predicted aroma impact. Ale yeasts were fermented at 20ºC (68ºF) and lager
yeasts were fermented at 15ºC (59ºF).
 YEAST AROMA COMPOUNDS

Isoamyl Alcohol
Fusel alcohol, whiskey, cognac

Phenethyl Alcohol
Floral, rose, bread crust

Isoamyl Acetate
Banana, fruity

Phenethyl Acetate
Floral, honey

Ethyl Acetate
Grape, pear

Ethyl Hexanoate
Pineapple, tropical fruit

Ethyl Octanoate
Fruity, wine-like, apricot

Ethyl Decanoate
Red apple, cognac

Citronellol
Citrus, lemongrass, citronella

4-vinylguaiacol
Clove, black pepper
Key Takeaways
You can select specific yeasts for their flavour
characteristics.
Our diverse yeasts produce a diverse assortment of flavour
compounds. You can use the data to build the profile you desire.
For example, to push ethyl hexanoate (pineapple-like), you could
select Ardennes Ale or Hornindal Kveik Blend.

When making hoppy beers, look for yeasts with

more terpene biotransformation (beta citronellol) to
optimize impact of survivable aroma compounds.
Higher production of beta-citronellol indicates stronger terpene
biotransformation by the yeast, which may enhance the aromatics
of your kettle, whirlpool, and early dry hops. For example, you
could select Hydra or Cerberus to push more citrus aromatics via
hop terpene biotransformation in an IPA.

Production of volatile phenols is an important factor

in which style the strain is appropriate for.
These spicy, clove-like aromatics are out of place in certain styles.
While some yeast strains are quite strong producers of fruity
flavour compounds (e.g. Saison Maison), phenolics can have a
“masking” effect on fruity flavours. Phenolic strains best suit
specialty Belgian, Wit, Weizen, and Saison styles. To push flavour
to new heights in a Witbier, look toward Fruity Wit.

16
 YEAST AROMA COMPOUNDS
BY YEAST CATEGORY

Non-Phenolic Yeasts
(American, British, Kölsch, Kveik)

LEGEND

Non-phenolic yeasts are used to produce most beer styles out there, including most
American and British styles as well as speciality styles such as Kölsch. This group has a
wide range of flavour profiles and intensities for brewers to select to match to their recipe.
American Ale can be selected to get some biotransformation with minimal esters. House
Ale (available in dry yeast format) offers a balanced and mild profile. Hydra, Cerberus,
Hornindal Kveik, and English Ale II are the flavour and biotransformation powerhouses of
this group.

Note: these fermentations were conducted at 20ºC, but many of the Kveik strains can be
much more expressive at higher temperatures.

Wild yeasts produce phenolics, and loss of phenol production is a result of yeast
domestication by humans - just like how domesticated dogs have changed from wild
wolves. These yeasts have lost this “wild” trait since the genes responsible for
phenol production are inactivated. This is like how your Pug probably doesn’t howl at
the moon or travel in packs!

17
 Phenolic Yeasts
(Saison, Wit, Weizen, Wild)

LEGEND

Phenolic yeasts are also known as “Phenolic off flavour positive” or “POF-
positive” since this flavour is considered a flaw in some beers. These yeasts are
used to produce a wide range of beer styles including most Belgian ales,
saisons, hefeweizens, and wild ales. These yeasts are defined by their
production of 4-vinylguaiacol, which imparts a spicy, clove note to the beer.
Many brewers will identify this flavour as distinctly Belgian. These yeasts are
also some of the most intense out there, able to push huge amounts of esters
and make absolute flavour bombs.

Yeast isoamyl acetate (banana) production depends on the amount

of glucose in your wort. Struggling to get enough banana in your
hefeweizen? Try adding glucose (dextrose) to the wort!

18
 Lagers
S. pastorianus

LEGEND

Compared to ale strains (mostly S. cerevisiae with some interspecies hybrids),

lager yeasts (S. pastorianus) are comparatively more cold-tolerant and tend to
be more neutral in flavour profile. Brewers can select strains with more
expressive profiles for full-flavoured lagers, and strains with lighter profiles for
ultra clean and crisp summer beers.

Lager yeast is a hybrid of two yeast species, S. cerevisiae (ale yeast)

and S. eubayanus (a cold-tolerant Saccharomyces yeast). S.
eubayanus wasn’t discovered until recently and now is being used
to develop brand-new lager hybrid strains.
 With this
knowledge, you
can select the
right yeast for
your desired
profile.
WHAT IS
BIOTRANSFORMATION?

Biotransformation Brewers are intrigued about

biotransformation for several
is the process by reasons. Firstly, it offers an
which yeasts change opportunity to be creative with
less desirable or craft beers as they develop
recipes to add new flavours
flavourless and aromas. Yeast
compounds into more biotransformation also offers
desirable and bold opportunities to unlock
efficiencies in the use of other
flavours and aromas. ingredients such as hops.

21
In other words,
flavours from
your other
ingredients
are brought to
life by yeast!
TERPENE
BIOTRANSFORMATION

An example of biotransformation of terpene compounds by yeast. The yeast enzyme NADPH

dehydrogenase transforms hop terpenoids such as geraniol into citrusy-tasting citronellol.

What are hop terpenes?

Of all plants, hops is the species with the most complex known essential oils.
Hundreds of compounds have been identified in hop essential oils but the
composition and amount of these compounds depend on the hop variety.

The most popular class of compounds present in hop oils are terpenoids.
Terpenoids possess strong sensory qualities that can stick around in your beer.
This has led to them being called “survivable hop compounds”. Most of the
terpenoids important in beer are monoterpene alcohol, which we will call
"terpenes" from this point onward.

Which terpenes can be biotransformed by yeast?

An example of a common terpene in hops is geraniol which can be transformed
into other compounds including [beta] citronellol (citrus), nerol (lemon-lime) and
linalool (fruit loops). Alpha-humulene, another terpenoid compound, gets
converted into caryophyllene, which can add a note of black pepper to beer.

23
 Yeast can convert terpenes into other forms during active fermentation, including citronellol which is
not naturally found in hops. This is one of the primary methods of yeast biotransformation in beer!

How does beta-glucosidase activity factor into terpene biotransformation?

Beyond transforming terpenes into other forms, yeast can also release
glycosidically bound terpenes using a beta-glucosidase enzyme. This releases a
sugar molecule from the glycoside and a flavour-active terpene molecule. While
the beta-glucosidase activity of most brewing yeast strains is fairly minimal,
some strains of Brettanomyces have very strong beta-glucosidase activity!
Brewers can also use purified beta-glucosidase enzyme as a processing aid.

Which hop varieties are rich in terpenes?

Luckily, scientific publications and industry resources share knowledge of
terpene concentrations of various hops. Try Cascade, Chinook, Mosaic,
Centennial, Strata, and Bravo as a starting point for terpene-rich hops.

When should a brewer saturate their wort with terpenoids?

It is best to saturate your wort with terpenes in the late boil and whirlpool. We
recommend a starting point of 0.4kg per hL (1 lb per bbl) of terpene-rich hops
added to the whirlpool. This will maximize the solubility of the terpenes and
expose the yeast to them for the entire duration of fermentation.

How do I maximize terpene biotransformation impact in an IPA?

We recommend pairing a non-phenolic yeast strain such as Hydra or Cerberus
with terpene-rich hops such as Mosaic and Strata as a whirlpool addition. We
recommend non-phenolic yeasts for this application because phenols can mask
fruity flavours.

Yeast strains can be selected to enhance the

impact of hop terpene biotransformation.

24
 Terpene Biotransformation:
Beta Citronellol Production

English Ale I
Laerdal Kveik
Spooky Saison
Voss Kveik
Mexican Lager
St. Remy Abbey Ale
Copenhagen Lager
Citronellol is used as our
Czech Lager
Premium Pils marker for terpene
Isar Lager biotransformation by yeast,
Cali Ale because it is not naturally
Arset Kveik Blend present in hops!
Biergarten Lager
Autobahn Lager
Foggy London Ale
Saison Maison
Kölsch Ale
Anchorman Ale
Old World Saison Blend
House Ale
Irish Ale
Ebbegarden Kveik Blend
Ardennes Ale
French Saison
Fruity Wit
American Ale
Classic Wit
American Ale
Classic Wit
JÖTUNN
Vermont Ale
West Abbey Ale
English Ale I
Weizen I
Hornindal Kveik Blend
Dry Belgian Ale
Hydra
Cerberus
Uberweizen
Farmstand Saison

0 2 4 6 8 10
Sensory Threshold
THIOL
BIOTRANSFORMATION

What are thiols?

Thiol compounds can smell like Sauvignon Blanc wine, gooseberry, guava,
grapefruit, and passionfruit! It’s no surprise that the hops with the highest
amounts of free aromatic thiols are also the most expensive: Citra, Sabro,
Mosaic, Nelson Sauvin, to count a few. Thiol compounds have in common a
sulfur-hydrogen group, which tends to make these molecules really noticeable
to humans.

Which thiols are found in beer?

This list is not exhaustive, but illustrates

some of the key compounds when we talk
about thiol aromas in beer. These
molecules contribute tropical fruit notes to
beer.

How does thiol release work?

Thiols and their release is all the rage these days, with new yeasts coming onto
the market that have been bioengineered or bred to enhance thiol release. This
includes our Thiol Libre strain, which was developed by selective yeast
breeding. Thiol liberating yeast strains open up a new world of opportunities for
flavour development, and give us entirely new ways to make aromatic beers.
Thiol-releasing yeasts have been developed to have enhanced activity from the
IRC7 enzyme, which is a cysteine beta lyase that helps the yeast release
aromatic thiols from flavourless precursors (cysteine-bound thiols).

27
What can I do with thiol releasing yeasts?
With hops, you tend to pay a lot of money for unbound thiols because they
make hops smell intense. However, there is huge opportunity to release bound
thiols from lower-cost ingredients, in unexpected places. Some common and
lower-cost hops such as Cascade, Saaz, and Calypso contain large amounts of
bound thiols that can be released by yeasts with strong thiol release capabilities.

How do I release thiols in beer?

Select a yeast such as Thiol Libre that has strong beta lyase activity.

Select hops with high amounts of bound thiols, such as Cascade, Saaz,
Calypso, and Perle. This opens up the possibility of using and enhancing
locally-grown hops! Use these in the mash (mash hopping) and whirlpool.
You can dry hop with the big-money hops like Citra and Mosaic later on if
you like!
Try adjuncts like Phantasm that contain a high amount of bound thiol
precursors.
Experiment with base malts, including locally-produced options. Different
malts have different bound thiol concentrations.

Thiol release potential in Escarpment Labs strains

Aroma intensity of Escarpment Labs yeast strains grown in the presence of cysteine-bound 4MMP. Red
denotes low capacity for thiol release and green denotes higher capacity for thiol release. The wild
yeast YJM1400 isolated from guava fruit showed stronger release of cysteine-bound 4MMP compared
to the selected beer yeasts. This strain was selected for breeding with Ebbegarden Kveik to produce
Thiol Libre.

Screening yeasts for thiol (3MHA) and

ester (ethyl hexanoate) production in a
wort fermentation. The hybrid strain Thiol
Libre produced more 3MHA than its
parental strains. This strain also retains
the fruity ester production of its parental
strain Ebbegarden Kveik.

LEGEND
CONTROLLING
PHENOLICS
Production of phenolic aroma compounds is yet
another example of biotransformation by yeast.
However, this transformation is not desirable in
every beer style.

How does yeast make phenols?

Yeast makes volatile phenols such as 4-vinylguaiacol (clove) from phenolic acid
precursors such as ferulic acid in your wort, using enzymes called PAD1 and
FDC1. Different base malts and crop years can result in different amounts of
phenolic precursors. This means that malt selection can impact the amount of
phenol aroma you get from POF+ (phenolic) yeasts. This is important to
consider since the presence of phenolic flavours is a large driver of overall
beer flavour.

Production of volatile phenolics is another example of biotransformation

by yeast. However, it is not always desirable in the final beer.

How do I control yeast phenols?

This is a tricky one, because ferulic acid (and other phenolic acids) is impacted
by plant stress. This means that the phenol potential of malt can actually
change from year to year! The increased plant stress seems to typically
increase the total phenolic acid concentration of the grain, so it is possible that
more stressed barley results in more ferulic acid. As a result the amount of
ferulic acid may change from year to year and location to location.

How can I predict phenol potential of my malt?

We suggest doing a paired forced fermentation test using a French Saison
(POF+) and a California Ale (e.g. Cali Ale) strain by pitching the yeast at 4x the
usual inoculation rate into two separate flasks of wort prepared from the malt
you are testing, on a stir plate for 72 hours. A sniff test of the POF+ sample
should reveal the phenol potential of the malt you are using. Currently,
phenolic acid content is not a standard feature of malt specification sheets.
How does malt production and kilning impact phenol precursors?
During the kilning process of malts, the free phenolic acid in the husk will start
to polymerize with other compounds in the husk of the grain. This will in turn
make the phenolic acids insoluble in water which reduces the amount is then
extracted into the wort in the brewing process. This will ring true whether it is
American or European malt.

As a result, phenol-variable yeasts like Wit yeasts will be highly responsive to

the free PA content in the malt. To reduce phenols with phenol-variable yeasts,
we suggest an English or North American pale ale malt as your base. To
enhance phenols with Wit and Weizen yeasts, we suggest using a North
American or European Pilsner malt as a base malt.

How do I reduce phenolics when using a POF+ (phenolic) yeast strain?

Use Pale Ale malt rather than Pilsner malt as your base.
Avoid releasing ferulic acid into the wort by avoiding a ferulic acid rest
step during mashing.
Avoid trub carryover in your wort.
Select a temperature and pitch rate that optimizes production of fruity
esters.

30
IMPACT OF
TEMPERATURE ON YEAST
FLAVOUR PRODUCTION

ACETATE ESTERS

FATTY ACID ESTERS

Production of volatile phenolics is another example of biotransformation

by yeast. However, it is not always desirable in the final beer.

31
How do I maximize (or minimize) fruity esters in my beer?
While every strain is a bit different, we find that acetate esters such as isoamyl
acetate (banana) are often more present at the medium to low end of the
yeast’s fermentation temperature range. Never tried fermenting a Hefeweizen
at 16ºC? Try it!

On the other hand, fatty acid esters such as ethyl hexanoate (pineapple) tend
to be maximized around the yeast’s optimal growth temperature, which is
typically between 25-35ºC depending on the yeast. While we do not suggest
fermenting most strains above 25ºC, you can ferment saison yeasts and Kveik
yeast at elevated temperatures to maximize fatty acid ester production.

How do I minimize fusel (boozy) flavours in my beer?

We suggest avoiding temperatures over 25ºC for most strains other than
saison and Kveik yeasts, because above this temperature there is an elevated
risk for fusel (boozy) off-flavours in your beer.

What about phenolics and temperature?

The common lore is that yeast will produce more phenolics at higher
temperatures. Interestingly, we haven’t seen much evidence for this. In general,
yeast phenol production is more a factor of the amount of precursor extracted
from the malt. In fact, one study of German wheat beer showed lower levels of
styrene (a phenol) at elevated temperatures.

32
IMPACT OF YEAST PITCH
RATE ON FLAVOUR
PRODUCTION

What happens to the flavour if I under-pitch yeast?

Individuals - PCA Variables - PCA

These are PCA plots which show a “map” of the different flavour profiles of the yeast strains and
pitching rates. Samples near the top correspond to beta citronellol (biotransformation) and isobutanol
(fusel), while samples near the right correspond to esters (fruity).

In 2019, we put this question to the test! At the time, we were looking at kveik
yeast but we also included an ale strain popular for IPAs (Vermont Ale). We
tested multiple yeast pitch rates (1 million cells/mL, 7 million cells/mL, and 10
million cells/mL). The highest pitch rate is a “standard” rate for Escarpment
yeast supply, so the lowest rate represents a 10x underpitch!

For some strains (Ebbegarden and Arset) we didn’t see a noticeable flavour
trend with pitch rate. However, for Voss, Hornindal, and Vermont, we did see
stronger flavour production as the pitch rate decreased. This may be beneficial
for kveik as they trended toward higher concentrations of fruity esters.
However, it is likely not beneficial to under-pitch Vermont as this was
associated with more fusel alcohol (isobutanol).
33
SOUR BEERS: FLAVOUR
FROM LACTOBACILLUS

These are PCA plots which show a “map” of the different flavour profiles of Lactobacillus strains when
copitched with yeast. Plot A includes analytically measured values and Plot B includes values
measured using sensory analysis. For example, samples to the left correspond to acetic flavours while
samples to the right correspond to tropical fruit flavours. This shows the flavour diversity of
Lactobacillus strains!

Wait! I can get flavour from Lacto?

Yes! Lactobacillus
Lactobacillus used to make sour beers is not just a way to make the beer
sour - it also produces unique flavour profiles. Different strains and species of
Lactobacillus can drive completely different flavour profiles.

For example, Lactobacillus

Lactobacillus brevis
brevis is associated with acetic and funky flavours,
making it suitable for some barrel-aged and wild ales as well as traditional
Berliner Weisse where a complex acid profile is desired. Meanwhile,
Lactobacillus plantarum
plantarum is quite diverse with some strains associated with
fruity and citrusy flavour profiles. Lactobacillus
Lactobacillus rhamnosus
rhamnosus is associated with
fruity, tropical flavour notes. This led to the creation of our Lactobacillus
Lactobacillus Blend
2.0 which blends L. L. plantarum
plantarum and L. L. rhamnosus
rhamnosus to really push the citrus and
tropical fruit notes.
34
BRETTANOMYCES:
THE ULTIMATE ESTER BOMB

Brettanomyces is a very special yeast, known for its delightful range of

flavours which include both candied-fruit esters and bombastic funky barnyard
phenols. Brett can produce a dizzying range of esters, which can make
different strains taste like plum, pineapple, strawberry, cherry, and all sorts of
hypothetical fruit in between. Those phenols are also important, giving Brett a
potent complexity from their barnyard, hay, and campfire qualities. Those
phenols are also why Brett has such a bad reputation, especially in the wine
world. In some environments like barrel aging red wines, Brett can run amok
and produce excessive phenols and steamroll more delicate flavours.

Several Brettanomyces
Brettanomyces species have been classified including Brettanomyces
bruxellensis and Brettanomyces
Brettanomyces anomalus
anomalus . There are a few other species but
they don't show up in beer very often.

In general, B.
B. bruxellensis
bruxellensis is the most aggressive fermenter of the family, with
some strains capable of fully attenuating wort in primary fermentation. Some
strains are incredibly aromatic, producing up to three times as much in terms of
total flavour production compared to the most expressive Saccharomyces
Saccharomyces
strains.

They also tend to produce a higher quantity of funky phenols than other
species, so B. bruxellensis
bruxellensis tends to be your go-to for funk bombs. B. anomalus
is often more subtle and may lend citrus, peach, white wine, and lighter funk to
a beer. B.B. anomalus
anomalus strains have been shown to have strong beta-glucosidase
activity, making them highly suitable for biotransformation of hops.

LEGEND

Brettanomyces (Brett for short) is capable of

producing massive amounts of flavour in
beer primary fermentations. However, this
comes at a tradeoff with less reliable
fermentation and smoky/medicinal
phenolics such as 4-ethylguaiacol.
 ESCARPMENT LABORATORIES
BRETT STRAIN CATALOG
Primary
Product Species Flavours Suitable Styles
Ferment

Pear, tropical, Pale ales, sours,

Brett B B. bruxellensis
fruit Orval, clones

Hoppy beers,
Brett D B. bruxellensis Pineapple, hay Yes
pale sours

Fruit salad, Pale sours,

Brussels Brett B. bruxellensis
candy, Lambic "Lambic"

Hoppy beers,
Brett C B. anomalus Citrus, mango anything barrel
aged

Flanders red,
Cherry, Yes, but
Brett L B. bruxellensis Oud Bruin, barrel
pleasant, acetic slow
aged sours

Balanced fruit
Brett M B. bruxellensis Anything
and funk

Strawberry,
Saison, barrel
Brett Q B. bruxellensis tropical fruit, Yes
aged sours
red fruit

Berliner Weisse,
Citrus, white
Berliner Brett I B. anomalus Saison, pale
wine
sours

Berliner Weisse,
Berliner Brett Peach, white
B. bruxellensis Saison, pale
II wine
sours

Mixed ferm
Saisons, barrel
MOTHERSHIP A bit of
Both aged sours,
BRETT BLEND everything
mixed ferm pale
ales
So, what are
we going to
brew next?
BEER IDEAS
Let's take a look at what we've learned and use it
to our advantage.
BEER IDEAS
So, what can you do with the information contained in this guide? We have some
ideas! What follows are recipe suggestions based on the data presented in this
guide as well as other ideas in brewing. Since every brewery (and brewer) is
different, we are suggesting some principles and combinations you can
experiment with in your own brewhouse and recipe design process.

01 Terpene biotransformation for citrus and

mango character in hoppy beers

You can select yeasts with stronger terpene biotransformation potential which
can unlock stronger citrus or fruity characteristics from many of the hop varieties
typically considered to be more piney, resinous, or “dank”. While our absolute
favourite for this application is Strata, you can also look to hop varieties
Columbus, Chinook, Centennial, as well as Vista. We find that when classically
“dank” hops are paired with a yeast capable of terpene biotransformation and
ester production, the end results create a beautiful and complex mango-like
character.

Suggested Yeast
Hydra

Attenuation: Phenolic:
65-72% Non-phenolic

Optimal Temp: Suggested Hops:

20–25°C (68–77°F) Strata or another hop
high in Myrcene and
Diastatic: Geraniol
No Suggested beer styles:
Flavour profile: Hop forward beers
including Hazy IPA and
Mango, stone fruit
Pale Ale

39
 02 Use neutral yeasts to let natural hop
flavours shine

Part of the reason that the Chico/Cali style ale strain is so popular is that it steps
out of the way and lets your hops or other ingredients shine through. Beyond
that strain there are many other options, such as our EL-D1 House Ale dry yeast.
This strain is fast and clean with minimal esters. but with an elevated terpene
biotransformation potential compared to Chico/Cali yeasts.

These tips apply to utilizing lager yeast in a Cold IPA, as well. We recommend
our Premium Pils which is an extremely efficient fermenting strain at 15ºC (59ºF)
with a balanced lager yeast profile that lets the hops sing.

Suggested Yeast
House Ale | EL-D1

Attenuation:
75-85%

Optimal Temp:
16–22°C (61–72°F)

Diastatic:
No

Flavour profile:
Natural, clean, balanced

Suggested Hops:
Late kettle and whirlpool with hops high in volatile survivable terpenoid
compounds (e.g. Mosaic, Centennial) and dry hop with hops high in free/unbound
thiols such as Citra, Sabro.

Suggested beer styles:

West Coast IPA and Pale Ale

40
 03 Enrich your wort for thiol
biotransformation

Thiol biotransformation has become a popular topic due to the potential to

release tropical fruit and white wine notes from previously hidden precursors. In
order to maximize the amount of thiol precursor available to your yeast, we
recommend using hops high in bound thiol precursors which include Cascade,
Calypso, and Motueka. The earlier, the better with solubilization of bound thiol
precursors, with some brewers going as far as mash-hopping.

For mash hopping, we recommend a starting rate of 3-5g/L (approx. 1 lb per bbl)
and factoring in the IBU extraction from the mash hops at a utilization rate of 30-
40%.

Suggested Yeast
Thiol Libre

Attenuation:
68-80%

Optimal Temp:
17–23°C (63–73°F)

Fermentation Rate:
High

Flavour profile:
Passionfruit, Guava, Grapefruit

Suggested Hops:
Late kettle and whirlpool with hops high in bound thiol precursors including Cascade,
Calypso, and Motueka. Perle and Saaz are also options. Balance this with a hop that
will round out the flavour, such as Cashmere or Chinook. When dry hopping, we
recommend starting at a lower rate and using hops that contain some unbound thiols
such as Citra, Sabro, or Simcoe.

Suggested beer styles:

Hazy/NEIPA, West Coast IPA, and Pale Ale

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04 Supercharge your wheat beers
While the Franco-Belgian styles aren’t exactly gaining in popularity these days,
we love our yeast-forward beers here at Escarpment Labs. And the yeasts used
to make beers like Saison, Hefeweizen and Witbier are some of the most
aromatic and capable of biotransformation.

Pairing a yeast with high biotransformation potential with the right hops and
adjuncts can help you fine-tune the profile of your beer. For example, you can
design a Witbier to pair Fruity Wit with hops high in geraniol such as Perle as
well as coriander (high in linalool), resulting in enhanced aroma presence.

Suggested Yeast
Saison Maison

Attenuation:
72-80%

Optimal Temp:
22–35°C (72–95°F)

Diastatic:
No

Flavour profile:
Pepper, Orchard Fruit

Suggested Hops/Adjuncts:
Since many Saison/Wit/Weizen yeasts are capable of noticeable terpene and thiol
biotransformation, you can experiment with European hops high in bound thiol
precursors such as Saaz and Perle. You can also get really interesting results from
“old school” American hops such as Mt Hood and Sterling or British hops such as
East Kent Goldings. Adjuncts such as coriander may also be biotransformed by yeast!

Suggested beer styles:

Saison, Witbier, Hefeweizen

42
 05 Make a really tasty sour
in no time at all
There are a lot of boring kettle/quick sours out there. We think the process can be
tweaked through microbe selection to yield much more flavourful results, including
thirst-quenching fruited sours and tart hoppy beers.
Here is our process:

Produce an unhopped wort base for kettle/quick souring, preacidifying to pH 4.5

using phosphoric or lactic acid.

Cool to 37ºC (99ºF) and pitch Lactobacillus Blend 2.0, which is selected to
enhance fruity characteristics in kettle/quick sours. This blend is also selected to
avoid hop-resistant Lactobacillus species. Sour until your desired pH, usually 18-
36 hours. It is OK if the temperature drops to 30ºC (86ºF).

Pitch a flavourful and high-temperature tolerant Kveik yeast alongside one dry
hop charge. The dry hop charge will inhibit the Lacto from producing more acid.
We recommend Laerdal Kveik or Hornindal Kveik Blend, both of which are
selected to offer a high presence of fruity ester compounds. For best results,
pitch either of these strains around 30ºC (86ºF).

Dry hop and/or fruit once a stable terminal gravity has been reached. Make sure
to monitor for a stable specific gravity before packaging.

Suggested Yeast
Laerdal Kveik

Attenuation: Suggested
72-80% Hops/Adjuncts:
Calypso, Equanot, Azacca
Optimal Temp:
22–35°C (72–95°F) Suggested beer styles:
Fruited Sour, Gose, Dry
Phenolic: Hopped Sour
Yes

Flavour profile:
Pineapple, Orchard
Fruit

43
 But wait!
What about dry hopping mid-ferment?
Isn’t that what biotransformation means?

Dry hopping mid-fermentation can offer some benefits in terms of

beer flavour, especially in hazy/New England style beers. So
much so, that this practice is often referred to as
biotransformation dry hopping. However, there is limited
evidence that this practice facilitates greater biotransformation
versus other approaches such as loading up the late kettle and
whirlpool with transformable compounds such as monoterpenes
and bound thiols.

Furthermore, many well-regarded producers of hazy IPAs avoid

the mid-fermentation dry hop entirely because it makes yeast
much harder to crop and repitch.

If you have been dry-hopping mid-ferment for the last few years,
we encourage you to experiment with shifting your first dry hop
toward the end of the fermentation and to ensure you include lots
of transformable compounds on your hot side. You may find this
improves yeast management by making the yeast much easier to
crop and repitch.

For many yeast strains, you can crop enough yeast for your next
batch on day 3 or 4 of fermentation without negatively affecting
diacetyl reduction or hop creep.
About this Guide
This guide was written and assembled by our co-founder Richard Preiss.

A number of researchers contributed to the data presented in this work:

Caroline Tyrawa, Iz Netto, Barrett Foster, University of Guelph co-operative

education students, Eugene Fletcher, the George van der Merwe lab at the
University of Guelph, and the Gibson/Krogerus group at VTT Finland.

Additional content in this guide was contributed by Eugene Fletcher and Phillip
Jurek.

This work would not have been possible without prior investment in public and
private scientific research. In particular, we would like to highlight the recent
efforts by VTT Finland, Omega Yeast, Berkeley Yeast, Imperial Yeast, Oregon
State University, University of Guelph, Nyséos, KU Leuven, White Labs,
Lallemand, and many more for advancing our understanding of yeast flavour
and biotransformation.
Learn More
Do you want to learn more about beer fermentation? Escarpment Labs
prioritizes creating and sharing our knowledge. Check out these resources:

Our
Our blog
Blog (www.escarpmentlabs.com/blog), with more than 100 informative
posts including how to do a Forced Diacetyl Test and how to ensure
success with Kveik yeast.

Our
Our Knowledge
KnowledgeBaseBase (www.escarpmentlabs.com/knowledgebase) with
over 80 entries covering topics including Yeast Pitch Rate, Yeast FAN (free
amino nitrogen) Requirements, and many Troubleshooting articles.

Our YouTube
YouTube Channel
Channel (www.escarpmentlabs.com/youtube) with more
than 20 hours of educational brewing content and brewer interviews
including our Yeast Basics series (Part 1 and Part 2).

For information on precursor compounds such as terpenes and thiols in hops,

we recommend the following resources:

Hop
Hop Oils
Oils and
andAcids
AcidsRankings
Rankings by Scott Janish (https://scottjanish.com/hop-
oils-and-acid-rankings/)

BeerMaverick hop database and hop substitution chart

(https://beermaverick.com/hops/)

Survivable
SurvivableCompounds:
Compounds:AABrewer’s
Brewer'sHandbook.
Handbook by YCH Hops

Blog posts by Omega Yeast and Berkeley Yeast

Hop Queries newsletter by Stan Hieronymus

(https://appellationbeer.com/blog/hop-queries/)

46
The Nerdy Details

If you are curious about the methods used to produce the data in this Guide,
here is some information:

Yeast aroma profile data were collected using HS-SPME-GC-MS on beer

samples in triplicate. Averages of triplicate data are shown. The wort was a
12.5ºP original gravity Pale Ale wort using Magnum and Cascade hops. Ale
strains were fermented at 20ºC (68ºF) and lager strains were fermented at
15ºC (59ºF).

Data on thiol release potential and thiol production were collected using
HS-GC-FID (non-thiol compounds) and HS-SPME-GC-MS/MS (thiol
compounds). Aroma intensity was based on growing the yeast in presence
of cysteine-bound 4-MMP and measuring aroma intensity by trained
sensory professionals. This project was published in the journal Applied
Microbiology & Biotechnology in 2021.

Impact of temperature on yeast flavour production data were collected

using HS-SPME-GC-MS on beer samples collected in triplicate. Averages of
triplicate data are shown. Results from this project were shared at World
Brewing Congress 2020 (virtual).

Impact of yeast pitching rate on flavour production data were collected

using HS-SPME-GC-MS on beer samples collected in triplicate. Averages of
triplicate data are shown. Results from this project were shared at MBAA
Conference 2019 (Calgary, Canada).

Lactobacillus impact on beer aroma data were collected using HS-SPME-

GC-MS on beer samples collected in triplicate. Averages of triplicate data
are shown. Sensory analysis was also performed using qualitative
descriptive analysis (QDA) by 16 trained tasters. Results from this project
was shared at World Brewing Congress 2020 (virtual).

If you would like a copy of any of the scientific papers, posters, and slideshows
referenced above, feel free to email help@escarpmentlabs.com.

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49
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Guelph, ON | N1C 0A1
Canada

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info@escarpmentlabs.com