Art of Brewing

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
Kindly provided by Tom Pugh and David Ryder of Miller Brewing Company
The Art of Brewing
Definition of Beer
An alcoholic beverage produced by the

fermentation of sugar-rich extracts derived
from cereal grains or other starchy
materials.
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
History of Brewing
Sumarian beer recipe

3000 BC
Resembled liquid bread:
Barley and Emmer
Spices / fruits
No Hops
Safe, nutritious, and
exhilarating beverage.
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.
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 . . .
.small animal which sips sugar through its snout, and
excretes alcohol from its gut and carbonic acid from its
urinary organ.
1866 - Louis Pasteur

Yeast was responsible for alcoholic
fermentation.
1883 - Emil Christian Hansen

Developed pure culture technique
Isolated pure cultures of brewing yeasts
Brewing Yeasts
Types of Brewing Yeasts
Two types of brewing yeasts, originally classified

on flocculation behavior
Top-fermenting
Ale yeast
Weiss yeast
Bottom-fermenting
Lager yeast
Weiss Ale
Lager Lab
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.
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.
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.
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
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 + - +
Distinguishing Characteristics
Difficult to distinguish between different lager yeasts
using conventional techniques
Colony and cell morphologies similar
Fermentation characteristics
PCR - limited success
Electrophoretic karyotyping
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
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
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
The Brewing Process
Ingredients
Malted barley
Cereal Adjunct
Hops
Water
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
Steeping
Soak, aerate, drain.

2 days
Germination
Ventilated to remove CO2

Repeated turning
4 to 5 days
Cereal Adjuncts
Types of adjuncts commonly used:

Corn grits
Rice
Corn syrups (high maltose and dextrose)
Purpose:
Additional source of fermentable sugars
Lighter body
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
Hops
The Brewing Process
Step Purpose
Brewhouse Starch Sugars
Wort production
Fermentation Sugars Ethanol

Flavor production
Carbonation
Lagering
Flavor maturation
The Brewing Process
Malt Mill
Cereal
Mash Tun
Cooker Brink Fermentation
Lauter Tun
Brew Hops
Kettle Aeration
Hot Wort Lagering

Receiver
Wort
Cooler
Mash Tun / Cereal Cooker
Activate malt enzymes
Convert starch to
fermentable sugars
Lauter Tun
Strainer
Brew Kettle
Sterilization
Protein coagulation
Hop extraction
Volatile removal
The Brewing Process
Malt Mill
Cereal
Mash Tun
Lauter Tun
Brew Hops
Kettle Aeration
Hot Wort Lagering

Receiver
Wort
Cooler
Wort Composition
Carbohydrates
20
73% Fermentable
15
11.77
10
CHO (% w/v)
4.43
5
0
Fermentable Non-fermentable
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
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)

The Brewing Process
Malt Mill
Cereal
Mash Tun
Lauter Tun
Brew Hops
Kettle Aeration
Hot Wort Lagering

Receiver
Wort
Cooler
Fermentation
Yeast growth Flavor compounds
Alcohol and CO2 Large - 600,000 L
Lagering
Carbonation Off-flavor reduction
The Brewing Process
Malt Mill
Cereal
Mash Tun
Cooker Brink
Fermentation
Lauter Tun
Brew Hops
Kettle Aeration
Hot Wort Lagering

Receiver
Wort
Cooler
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
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
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

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

Vicinal Diketones
Threonine Pentanedione
-ketobutyrate -acetohydroxybutyrate Isoleucine
pyruvate -acetolactate Valine
Diacetyl
Buttery, butterscotch flavor
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.

Art of Brewing

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Art of Brewing

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The Art of Brewing

Provide a better understanding of...

The brewing process

Types of brewing yeasts

Attributes important to the brewer

An alcoholic beverage produced by the

Man has been making beer since the dawn of

Sumaria (4000 BC) Sikaru

Sumarian beer recipe

Unwittingly, ancient brewers domesticated yeast.

Deduced that yeast was important to make beer.

Did not know what yeast was.

1680 Antonie van Leeuwenhoek

1837 - Cagniard Latour

1839 -Justus von Liebig and Friedrich Wohler

1866 - Louis Pasteur

1883 - Emil Christian Hansen

Two types of brewing yeasts, originally classified

Strains are genetically more diverse - several origins

Warm fermentation temperatures: 65 to 72 F.

Produces beer that has spicy, clove, vanilla, and

Warm fermentation temperatures: 65 to 72 F.

Ale and Weiss yeasts - Saccharomyces cerevisiae

PCR - limited success

Allopolyploid and probably aneuploid.

Contains two types of chromosomes

Gene length similar, but nucleotide divergence.

Gene Nt. Identity AA. Identity

Soak, aerate, drain.

Ventilated to remove CO2

Types of adjuncts commonly used:

Fermentation Sugars Ethanol

Hot Wort Lagering

Activate malt enzymes

Hot Wort Lagering

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

Hot Wort Lagering

Hot Wort Lagering

The brewer needs...

Oxygen is growth limiting nutrient.

Alcohol Amino Acid -keto acid

Amyl Isoleucine -keto-3-methylvalerate

Isobutanol Valine -keto-isovalerate

Propanol Threonine -keto-butyrate

Alcoholic, solventy, and fruity flavor notes

Fruity flavor notes

-ketobutyrate -acetohydroxybutyrate Isoleucine

pyruvate -acetolactate Valine

Buttery, butterscotch flavor

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