Institute of Brewing and Distilling

General Certificate in Brewing (GCB)

Section 6
The Basic Principles of Yeast Fermentation.

6.1 Brewing Yeast:-

What it is.

Yeast is a single celled micro-organism, in fact it is a minute fungus

which is larger than any bacteria.

A yeast cell

Bud scar
Cell wall

Cytoplasm
Mitochondrion

Vacuole

Nucleus

Under the microscope, the appearance of the cells gives information

about the yeast.

• Bud scars indicate its age.

• The shape and density of the vacuole indicate phases in its growth.
• The size of the cell gives an indication of the strain. There are
numerous strains of yeast used in brewing, many having characteristics
that create unique flavours during fermentation.
• The presence of chains of cells indicates the class of yeast. Yeast is
classified depending on how it performs during the fermentation.

Yeast cells multiply by ‘budding’ as shown in the diagrams below:-

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 1. 2. 3.

New bud
New cell scars

How yeast lives.

Unlike green plants, yeast is unable to utilise sunlight so it has to make

use of simple sugars to satisfy its requirements for energy and growth. It
can utilise sugars in the presence or absence of oxygen.

When oxygen is available to the yeast it respires aerobically and sugars

are converted into carbon dioxide and water and energy is created to
produce new cells. Some unpleasant by-products are also produced.

When there is no oxygen available, the yeast respires anaerobically and

the by-products from sugar utilisation are alcohols (mainly ethanol),
carbon dioxide and the range of aromatic substances that make up a
pleasantly flavoured beer. This process is called fermentation.
Anaerobic respiration does not generate much energy and new cells are
not produced.
Nearly all of the sugars formed from the barley starch during mashing can
be used by yeast; sugar fermentability is described in Section 2.3.

Yeast Nutritional Requirements

Yeast has nutritional requirements other than just sugar and these are
listed below:-

• Protein or nitrogenous compounds in the form of amino acids. These

are created from the barley protein during malting and mashing.
• Lipids or fatty material. This is also supplied by the malt.
• Vitamins from the malt.

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 • Trace metals. Calcium is usually present in the brewing water, if not it
must be added as described in module 6.1. Zinc may be present in hop
products, if not it can be added to the wort.
• Oxygen is usually dosed into the wort. Oxygen is essential for healthy
yeast growth and a large yeast population is required to ensure that the
fermentation is healthy and fast.

Different types of yeast.

There are two main types of yeast used in brewing:-

Saccharomyces cerevisiae Saccharomyces carlsbergensis

• The ‘top’ yeast used for
fermenting ales.
• It floats to the top of the vessel at • It sinks to the base of the vessel at the
the end of a fermentation because
the carbon dioxide bubbles stick to
the yeast’s cell walls.
• It thrives on relatively high
fermentation temperatures, for
example 20°C and consequently
fermentations are fast, for example
3 days.
• The flavours produced by
fermentation are typical of ale
flavours.
• The system used for cropping the
yeast at the end of fermentation,
that is skimming the yeast off the
top of the beer, naturally selects
the best yeast for repitching.
• Beer containing this yeast can be
clarified by the addition of finings. • Beer containing this yeast cannot be
• It cannot ferment a sugar called
‘melibiose’.
(or uvarum)
• The ‘bottom’ yeast used for
fermenting lagers.
end of a fermentation because it has a
different kind of cell wall.
• It likes low fermentation temperatures,
for example 10°C and fermentations
are slower, for example 7 days.
• The flavours produced by
fermentation are typical of lager
flavours.
• The system used for cropping the
yeast at the end of fermentation, that is
collecting from the base of the vessel
is not selective and usually a pure
culturing system is in use to maintain
yeast purity.
clarified by the addition of finings.
• It can ferment a sugar called
‘melibiose’.

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Within each type of yeast there are numerous strains with each strain
performing differently in terms of which sugars it can ferment’ how
effectively it settles out after fermentation and what flavours it produces.

For this reason, many beer brands have their own specific pitching yeast.

Some breweries deliberately have more than one strain in their pitching
yeast a concept that works in their specific conditions.

Yeast Pitching.

Pitching is the term used for adding yeast to the wort to start the
fermentation.
The choice of pitching yeast has a major influence on the performance of
the fermentation and its outcome.

A pitching yeast must have the following characteristics:-

• The right strain for the beer to be fermented.

• Free from contamination by bacteria and other yeasts.
• Healthy and viable.
• Cropped from a healthy and consistent fermentation itself. Yeasts
selected from a slow or sticking fermentation are likely to repeat the
problem.

Selection will be based on laboratory analysis and fermentation records.

Notes.
Describe the yeast used in the fermentation of a beer that you are familiar
with.
Observe your brewery’s pitching yeast under the microscope and draw a
diagram.

6.2 Fermentation Theory

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Yeast activity.

How yeast grows, ferments and then settles out is illustrated in the
diagrams below:-

Four phases of fermentation

1. Pitching. 2. Yeast growth 3. The yeast 4. After

About 10 Yeast multiplies ferments the fermentation, the
million cells to a dense sugar into yeast floats to
per ml are population. alcohol. CO2 the top or settles
added to the and heat are to the bottom
wort. produced.

Fermentation.

The conversion of sugars into alcohol can be tracked by measuring the

specific gravity of the liquid. Alcohol is less dense (lighter) than sugar
and CO2 is lost, so specific gravity drops as the fermentation progresses.

This process is illustrated in the graph below:-

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 1

4
2 3
Specific gravity

Time

1. Nothing happens to the specific gravity until the yeast has been pitched
in. When the yeast starts to get used to its environment, it starts to grow.
This period is called the lag phase and its length depends on how much
yeast is added (the pitching rate) and how healthy it is.

2. The specific gravity drops slowly at first because the yeast is growing.
This is called the growth phase and how long it is depends on temperature
and most importantly, how much air/oxygen has been added to the wort.

3. The specific gravity drops rapidly during the fermentation phase. The
fermentable sugars are quickly converted into alcohol, large volumes of
carbon dioxide are produced and heat is generated. The pH of the beer
also drops during fermentation.

4. The drop in specific gravity slows down as the sugars are used up and
the yeast settles out of suspension. Top fermenting yeast may be cropped
at this stage. The beer may be reduced in temperature.

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6.3 Fermentation and Beer Flavour
Yeast takes in carbohydrates in the form of sugar to generate the energy it
needs for life. When no air is present, yeast converts those sugars into
alcohol and carbon dioxide by the process of fermentation.

Fermentable
CO2 CO2
Oxygen Sugar

YEAST
CELLS
Alcohol

Respiration Fermentation
'growth' 'no growth'

During the fermentation process, yeast converts the sugars produced in

the brewhouse into alcohol. The process starts when yeast is pitched into
the wort and finishes when most of the sugar has been converted into
alcohol and carbon dioxide.

The overall reaction for fermentation is:

C6 H12 O6 ---------► 2 (C2 H5 OH) + 2 (CO2) + Energy

Glucose (sugar) Ethanol (alcohol) Carbon Dioxide

This equation summarises very many metabolic reactions occurring

during fermentation, all controlled by a variety of different enzymes in
the yeast cell. The main purpose of the breakdown of glucose to alcohol
and carbon dioxide is to generate energy necessary for the yeast cells to
survive and grow. However, many of the metabolic reactions carried out
by the yeast, such as making proteins and fats for new cells, also produce
by products, which can be very important to beer flavour.

These by products, such as esters, higher alcohols, diacetyl and sulphur

compounds, can make major contributions to beer flavour and various
fermentation conditions can influence the amounts of these compounds.

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The yeast used for brewing beer is carefully selected because it influences
fermentation performance and the beer’s eventual flavour, by influencing
the amounts of various flavour compounds.

The various fermentation conditions that are important in this respect

include:

• Selection of pitching yeast.

• Pitching rate (the amount of yeast added to the wort). The amount of
yeast in suspension can be measured by ‘yeast count’.
Ales are pitched at approximately 9 million cells per ml,
Lagers at 14 million cells per ml.

• Wort dissolved oxygen (the amount of air/oxygen added to the wort).

• Initial temperature (wort temperature before the yeast is added).

• The rate of temperature increase during the growth phase.

• Top heat (the maximum temperature during fermentation).

• Final temperature (what temperature the beer is reduced to at the end).

• Fermentation vessel design / shape.

Consistent application and tight control of these parameters is required to

produce a fermentation of consistent speed so that the beer produced has
consistent quality and flavour characteristics.

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Beer Flavour Compounds

Trace compounds Ethanol (alcohol)

Carbohydrate
(eg sugar)
Carbon Dioxide

Amino acids Higher alcohols

Diacetyl
Vitamins
YEAST
Acetaldehyde
Trace Elements

Sulphur compounds

Minerals

Esters

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Alcohol (ethanol) itself only makes a minor contribution to beer flavour,
other than having a warming effect.

Esters are very important beer flavour compounds and several hundred
can be found in beer, although only a few are present in sufficient amount
to contribute significantly to beer, and together the flavours generated by
esters are described as “fruity” and “tropical fruit”.
These compounds are essentially formed by combination of alcohols with
organic acids, so that those present in the highest quantity, such as ethyl
acetate (which contributes “boiled sweet”, almost “solvent” flavour
character), are derived from ethanol (since this is by far the most
abundant alcohol). Another very flavour active ester is iso-amyl acetate,
which is usually present in high enough levels to taste (i.e. above its
“flavour threshold”) and tastes of “bananas” or “pear drops”.

Esters are formed as the rate of fermentation slows down (as the yeast is
fermenting the last of the sugar), when the yeast stops the synthesis of
fats and the amounts of individual esters and the total produced are
affected, in the main, by fermentation factors that affect yeast growth.

• Especially important is the strength of the original wort (i.e.

wort gravity), so that stronger worts usually produce very high
levels of esters and so, strong beers taste very fruity. In fact, it is
the proportion of fermentable sugars to the amount of
assimilable nitrogen (mainly amino acids derived from malt
proteins during mash conversion) that is the main driving force
in determining the levels of ester production.

• Other factors that encourage yeast growth, such as increased

dissolved oxygen at the start of fermentation, reduce ester
formation.

• Increased fermentation temperature encourages ester

production.

• However increased pressure during fermentation tends to

decrease yeast growth and thus reduces ester formation. This
means that fermentations in very tall vessels produce lower
levels of esters, because of the increased hydrostatic pressures.

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Higher Alcohols (also known as Fusel Alcohols) are produced as by
products from protein synthesis and have aroma and flavour effects such
as “alcohol” , “winey”. The main examples are iso-butanol and iso-amyl
alcohol. The total concentration of higher alcohols produced during
fermentation is directly related to amount of yeast growth, so that factors
increases yeast growth also favour increased production of higher
alcohols:
• Increased level of wort oxygen
• Higher levels of wort FAN (free amino nitrogen)
• Increased fermentation temperature
• Increased pressure during fermentation tends to decrease yeast
growth and thus reduces higher alcohol formation (like ester
synthesis), so that fermentations in very tall vessels produce
lower levels of higher alcohols, because of the increased
hydrostatic pressures.

Diacetyl gives a “toffee” or “butterscotch” flavour to beer. This character

is often acceptable in ales, but is usually unpleasant in lagers and lager
processing is designed to reduce the level of diacetyl to below its flavour
threshold - approximately 25 ppb or micrograms / litre (µgm/ litre).
Diacetyl is produced during fermentation, but yeast will reabsorb it (and
so remove it from beer) during a warm conditioning stage. Consequently,
it is very important (especially for lager brewing) to monitor, by chemical
analysis, the level of diacetyl towards the end of fermentation and during
maturation to ensure that the beer is not chilled too soon before the level
of diacetyl is controlled to achieve the final beer specification.
The desired beer diacetyl specification determines when the beer may be
moved from fermentation vessel to the conditioning phase, or chilled, or
centrifuged or filtered.
Factors affecting diacetyl production (and removal by yeast) are:

• The yeast strain

• Wort composition

• The type of fermentation vessel (open or closed)

• Fermentation conditions favouring yeast growth rate, such as

high temperatures and pitching rates, and an increased level of
wort oxygen.

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Sulphur Compounds make a significant contribution to beer flavour.
When in excess, they can give rise to unpleasant off-flavours so the
fermentation should be managed with the measures described below to
make sure the level remaining in the final beer is sufficient low to be
below flavour threshold. This is especially important for the more volatile
compounds such as hydrogen sulphide (H2S), which smells of bad eggs
and sulphur dioxide (SO2 ) which smells of burnt matches. Both of these
sulphur compounds are produced by yeast from sulphate and are by-
products in the synthesis of sulphur-containing amino acids.
The removal procedures include:

• Ensure sufficient evolution of CO2 to purge these volatile

compounds from the beer.

• Extended maturation time (to allow these sulphur compounds to

escape).

• More vigourous fermentation processes result in lower levels of

the volatile sulphur compounds.

• Do not allow settled yeast to remain the fermentation vessel for

too long, since this may encourage older yeast to break down
and release other unpleasant sulphur flavours (“cooked meat”,
“autolysed yeast”).

One other sulphur compound often found in beer, but is principally

derived from malt is Dimethyl Sulphide (DMS), which smells and tastes
of “cooked vegetables” or “tinned sweetcorn”.
This compound is rarely detectable in ales, because it is usually removed
during kilning to produce ale malts, but can be present at tasteable levels
in some lagers, although others are designed to have specification levels
of DMS below flavour threshold (approximately 30 ppb or micrograms
/litre or µgm /litre).
The level of DMS surviving into beer is very much determined by the
amount of DMS (and its precursor) surviving in malt after kilning and
also how much remains after wort boiling. Those lager brewers who
specify for a tasteable level of DMS in the finished beer will specify
desired levels in malt and will design the wort boiling conditions so that a
controlled level of DMS will remain in the finished beer. Otherwise malt
specifications and boiling conditions will be designed to ensure sufficient
removal of DMS to achieve the required low level in beer.

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One important point is that some yeasts can produce a small amount of
DMS during fermentation, which may be significant, although it is usual
to control DMS levels in beer by malt specification and boiling
conditions.

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