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The Art of Brewing

and
The Biology of Lager Yeast

Tom Pugh
Miller Brewing Company
Purpose

Provide a better understanding of...

The brewing process

Types of brewing yeasts

Attributes important to the brewer

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The Art of Brewing

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Definition of Beer

An alcoholic beverage produced by the


fermentation of sugar-rich extracts derived
from cereal grains or other starchy
materials.

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History of Brewing

Man has been making beer since the dawn of


civilization.
Where grain was grown, beer was made.

Sumaria (4000 BC) Sikaru


Egypt (3000 BC) Zythum
India (2000 BC) Sura
China (2000 BC) Kiu

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History of Brewing

Sumarian beer recipe


3000 BC
Resembled liquid bread:
Barley and Emmer
Spices / fruits
No Hops
Safe, nutritious, and
exhilarating beverage.

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The Role of Yeast in Brewing

Unwittingly, ancient brewers domesticated yeast.


Selected yeast that made good beer.

Deduced that yeast was important to make beer.


Collect the creamy foam or sediment from one brew.
Use it to pitch the next brew.

Did not know what yeast was.

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The Role of Yeast in Brewing

1680 Antonie van Leeuwenhoek


Observed yeast in beer.

1837 - Cagniard Latour


Microbe is responsible for alcoholic fermentation.

1839 -Justus von Liebig and Friedrich Wohler


Alcohol is produced by a chemical process in which dead and
decaying yeast participated.
Satired Latours theory in Annalen der Chemie . . .

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.small animal which sips sugar through its snout, and
excretes alcohol from its gut and carbonic acid from its
urinary organ.

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The Role of Yeast in Brewing

1866 - Louis Pasteur


Yeast was responsible for alcoholic
fermentation.

1883 - Emil Christian Hansen


Developed pure culture technique
Isolated pure cultures of brewing yeasts

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Brewing Yeasts

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Types of Brewing Yeasts

Two types of brewing yeasts, originally classified


on flocculation behavior

Top-fermenting
Ale yeast
Weiss yeast

Bottom-fermenting
Lager yeast

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Weiss Ale

Lager Lab

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Ale Yeast
Predominant brewing yeast prior to the mid-1800s.
Displaced by lager yeast

Strains are genetically more diverse - several origins

Warm fermentation temperatures: 65 to 72 F.

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Weiss Yeast
Bavarian origins - closely related.

Produces beer that has spicy, clove, vanilla, and


nutmeg flavor notes - POF.
PAD1 gene phenylacrylic acid decarboxylase
Decarboxylation of ferulic acid forms 4-vinyl-guaiacol,
which gives the characteristic clove flavor.

Warm fermentation temperatures: 65 to 72 F.

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Lager Yeast
Bavarian origin.
1400s in Munich - cool fermentations (selective pressure)
Taken to Pilsen and Copenhagen in 1840s
Pale malt, soft water, aromatic hops
Became very popular - displaced ale yeast
Popularity fueled by advances of Industrial Revolution
Steam power, refrigeration, railroads, pasteurization and
filtration technology
Strains are closely related - common origins
Cool fermentation temperatures: 42 to 52 F
Beers are more delicate, clean, drinkable, and less
aromatic.
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Taxonomy

Ale and Weiss yeasts - Saccharomyces cerevisiae


Polyploid and probably aneuploid.
Non-mating
Sporulates poorly and poor spore viability
Lager yeast - Saccharomyces pastorianus
S. cerevisiae
S. carlsbergensis
S. uvarum
Sporulates very poorly - poor spore viability

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Distinguishing Characteristics
Colony morphology
Microscopic appearance
Chain formation
Fermentation characteristics
Flocculation behavior / flavor compound profiles
Growth at 37 C
Melibiase
Electrophoretic karyotyping
Yeast 37 C Melibiase POF
Lager - + -
Ale + - -
Weiss + - +
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Distinguishing Characteristics
Difficult to distinguish between different lager yeasts
using conventional techniques
Colony and cell morphologies similar
Fermentation characteristics

PCR - limited success

Electrophoretic karyotyping

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Genome Structure - Lager Yeast

Allopolyploid and probably aneuploid.


Tetraploid

Natural hybrid
S. cerevisiae and S. bayanus
S. cerevisiae and S. monacensis

Contains two types of chromosomes


S. cerevisiae type
S. bayanus type
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Genome Structure
Gene order and function highly conserved
Single chromosome transfer experiments

Gene length similar, but nucleotide divergence.


Low levels of recombination between homeologues

Gene Nt. Identity AA. Identity


ILV1 86 % 96 %
ILV2 85 92
MET2 84 94
URA3 79 93
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Electrophoretic Karyotypes
cerev. parad.
Lager Ale bayan. Weiss Lab
Lab Lager Ale pastor.
XII
XII IV
IV
XV, VII
XV, VII XVI, XIII
XVI, XIII II, XIV
X
II, XIV XI
X
XI V
VIII
V, VIII

IX
IX
III

III VI
VI I
I

T C C
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The Brewing Process

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Ingredients

Malted barley

Cereal Adjunct

Hops

Water

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Malted Barley
Two types of barley
2-rowed
6-rowed
Provides fermentable sugars,
flavor, and color.
Malting process:
Steeping
Germination
Kilning
Purpose:
Activate enzyme systems
Preserve for brewhouse

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Steeping

Soak, aerate, drain.


2 days

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Germination

Ventilated to remove CO2


Repeated turning
4 to 5 days

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Cereal Adjuncts

Types of adjuncts commonly used:


Corn grits
Rice
Corn syrups (high maltose and dextrose)
Purpose:
Additional source of fermentable sugars
Lighter body

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Hops
Spice of beer
Provides aroma
and bitterness
Flower (cone) of a
vine-growing plant
Humulus lupulus
Female triploid
Used as:
Whole cones
Pellets
Extracts

Lupulin Glands

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Hops

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The Brewing Process
Step Purpose
Brewhouse Starch Sugars
Wort production

Fermentation Sugars Ethanol


Flavor production

Carbonation
Lagering
Flavor maturation

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The Brewing Process
Malt Mill

Cereal
Mash Tun
Cooker Brink Fermentation

Lauter Tun

Brew Hops
Kettle Aeration

Hot Wort Lagering


Receiver

Wort
Cooler
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Mash Tun / Cereal Cooker

Activate malt enzymes

Convert starch to
fermentable sugars

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Lauter Tun
Strainer

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Brew Kettle

Sterilization

Protein coagulation

Hop extraction

Volatile removal

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The Brewing Process
Malt Mill

Cereal
Mash Tun
Cooker Brink Fermentation

Lauter Tun

Brew Hops
Kettle Aeration

Hot Wort Lagering


Receiver

Wort
Cooler
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Wort Composition
Carbohydrates
20
73% Fermentable
15
11.77

10

CHO (% w/v)
4.43
5

0
Fermentable Non-fermentable

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Wort Composition
Fermentable Sugars
** need to adjust to normal wort
100

80

60 52.9

40
28.4
Percent (% w/v)
20 16.1
2.6
0
Maltose Glucose Maltotriose Fructose

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Wort Composition
Amino Acids (** adjust to normal wort)
300
269
250

200
151
150 132 126
PPM 107 105 110
89 93
100
65 63 68
53 49 56
50 30 31
17
0
Arg

Met
Phe
Asp

Leu

Asn
Thr

Gln
Tyr
Glu

Pro
Lys
Gly
Ala

Ser
Val
His
Ile

Not included: Cys (2 ppm) and Trp (50 ppm)


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The Brewing Process
Malt Mill

Cereal
Mash Tun
Cooker Brink Fermentation

Lauter Tun

Brew Hops
Kettle Aeration

Hot Wort Lagering


Receiver

Wort
Cooler
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Fermentation
Yeast growth Flavor compounds
Alcohol and CO2 Large - 600,000 L

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Lagering
Carbonation Off-flavor reduction

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The Brewing Process
Malt Mill

Cereal
Mash Tun
Cooker Brink
Fermentation

Lauter Tun

Brew Hops
Kettle Aeration

Hot Wort Lagering


Receiver

Wort
Cooler
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Balanced Growth
Yeast growth affects beer flavor.
Need balance between yeast growth and beer flavor.

The brewer needs...


Desired flavor profile in desired time.
Sufficient yeast crop for subsequent fermentations.

Oxygen is growth limiting nutrient.


Control point

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Yeast Metabolism During Fermentation
Sugars Oxygen

Membranes
Glucose

CO2
Energy Unsaturated Fatty Acids Esters
Pyruvate Sterols
Ethanol
Higher
TCA Alcohols
Amino Acids
Acetaldehyde Cycle
VDK
Organic Acids

Sulfur
Amino Acids Volatiles
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Higher Alcohols
Formed by the decarboxylation and reduction of
-keto acids.
From amino acid anabolism and catabolism.

Alcohol Amino Acid -keto acid


Isoamyl Leucine -keto-isocaproate

Amyl Isoleucine -keto-3-methylvalerate

Isobutanol Valine -keto-isovalerate

Propanol Threonine -keto-butyrate

Alcoholic, solventy, and fruity flavor notes


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Esters
Closely linked to lipid metabolism - growth.
Reaction of an alcohol and fatty acid intermediate

Acetate esters
Ethyl acetate solventy, fruity, sweet
Isoamyl acetate banana
Phenethyl acetate roses, honey, apple
Fatty acid esters
Ethyl caproate apple, aniseed
Ethyl caprylate apple
Isoamyl decanoate tropical fruits

Fruity flavor notes


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Vicinal Diketones

Threonine Pentanedione

-ketobutyrate -acetohydroxybutyrate Isoleucine

pyruvate -acetolactate Valine

Diacetyl

Buttery, butterscotch flavor

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Thanks to David Ryder of Miller Brewing Company
and Tom Pugh, formerly of Miller Brewing Company,
for providing this presentation to the Saccharomyces Genome Database
for dissemination to the yeast community.