Industrial Microbiology Lec 10

INDUSTRIAL
MICROBIOLOGY
Lecture X
Production of Wines and Spirits
GRAPE
WINE
PRODUCTION OF GRAPE WINE
Wine is by common usage defined as a product of the

normal alcoholic fermentation of the juice of sound
ripe grapes.
Vitis vinifera
The production of wine is simpler than that of beer in

that no need exists for malting since sugars are
already present in the fruit juice being used. This
however exposes wine making to greater
PROCESS IN WINE MAKING
CRUSHING OF GRAPES
Selected ripe grapes of 21 to 23 Balling are
crushed to release the juice which is known as
must, after the stalks which support the fruits have
been removed.
These stalks contain tannins which would give the
wine a harsh taste if left in the must.
The skin contains most of the materials which give
CRUSHING OF GRAPES
For the production of red wines the skins of
black grapes are included, to impart the
color.
Grapes for sweet wines must have a sugar

content of 24 to 28 Balling so that a residual
sugar content is maintained after
fermentation.
The chief sugars in grapes are glucose and

fructose; in ripe fruits they occur in about
CRUSHING OF GRAPES
Grape juice has an acidity of 0.60-0.65%
and a pH of 3.0-4.0 due mainly to malic and
tartaric acids with a little citric acid.
During ripening both the levulose content

and the tartaric acid contents rise.
Nitrogen is present in the form of amino

acids, peptides, purines, small amounts of
ammonium compounds and nitrates.
FERMENTATION
(i) Yeast used: The grapes themselves harbor a
natural flora of microorganisms (the bloom)
which in previous times brought about the
fermentation and contributed to the special
characters of various wines.
Nowadays the must is partially sterilized by the

use of sulphur dioxide, a bisulphate or a
metabisulphite which eliminates most
FERMENTATION
The yeast which is used is Saccaromyces
cerevisiae var, ellipsoideus (synonyms:
Sacch. cerevisiae, Sacch. ellipsoideus,
Sacch, vini.)
Other yeasts which have been used for

special wines are Sacch. fermentati, Sacch.
oyiformis and Sacch. bayanus.
FERMENTATION
Wine yeasts have the following
characteristics:
(a) growth at the relatively high acidity (i.e.,

low pH) of grape juice;
(b) resistance to high alcohol content

(higher than 10%);
(c) resistance to sulfite.
FERMENTATION
(ii) Control of fermentation
(a) Temperature: Heat is released during the

fermentations. It has been calculated that on the
basis of 24 Cal per 180 gm of sugar the
temperature of a must containing 22% sugar
would rise 52F (11C) if all the heat were
stopped from escaping.
If the initial temperature were 60F (16C) the
temperature would be 100F (38C) and
fermentation would halt while only 5% alcohol
has been accumulated. For this reason the
FERMENTATION
(b) Yeast Nutrition: Yeasts normally ferment the

glucose preferentially although some yeasts e.g. Sacch.
elegans prefer fructose.
To produce sweet wine glucose-fermenting wine yeasts
are used leaving the fructose which is much sweeter
than glucose.
Most nutrients including macro- and micro-nutrients are
usually abundant in must; occasionally, however,
nitrogeneous compounds are limiting. They are then
made adequate with small amounts of (NH4)2 SO4 or
FERMENTATION
(c) Oxygen: As with beer, oxygen is required in

the earlier stage of fermentation when yeast
multiplication is occurring. In the second stage
when alcohol is produced the growth is anaerobic
and this forces the yeasts to utilize such
intermediate products as acetaldehydes as
hydrogen acceptors and hence encourage alcohol
production.
FERMENTATION
(iii) Flavor development: Although some flavor

materials come from the grape most of it come from
yeast action.
The flavor of wine has been elucidated with gas

chromatography and has been shown to be due to
alcohols, esters, fatty acids, and carbonyl compounds,
the esters being the most important. Diacetyl,
acetonin, fusel oils, volatile esters, and hydrogen
sulfide have received special attention.
Autolysates from yeasts also have a special influence
AGEING AND STORAGE

The fermentation is usually over in three to
five days.
At this time pomace formed from grape
skins (in red wines) will have risen to the
top of the brew.
At the end of this fermentation the wine is
allowed to flow through a perforated bottom
if pomace had been allowed.
FERMENTATION
When the pomace has been separated from wine
and the fermentation is complete or stopped, the
next stage is racking.
The wine is allowed to stand until a major portion

of the yeast cells and other fine suspended
materials have collected at the bottom of the
container as sediment or lees. It is then
racked, during which process the clear wine is
carefully pumped or siphoned off without
disturbing the lees.
FERMENTATION
The wine is then transferred to wooden casks (1001,000 gallons), barrels (about 50 gallons) or tanks
(several thousand gallons).
The wood allows the wine only slow access to oxygen.
Water and ethanol evaporate slowly leading to air
pockets which permit the growth of aerobic wine
spoilers e.g. acetic acid bacteria and some yeasts.
The casks are therefore regularly topped up to prevent
the pockets.
In modern tanks made of stainless steel the problem
of air pockets is tackled by filling the airspace with an
inert gas such as carbon dioxide or nitrogen.
FERMENTATION
During ageing desirable changes occur in the wine.

These changes are due to a number of factors:
(a) Slow oxidation, since oxygen can only diffuse

slowly through the wood. Small amounts of oxygen
also enter during the filling up. Alcohols react with
acids to form esters; tannins are oxidized.
(b) Wood extractives also affect ageing by affecting
the flavor.
(c) In some wines microbial malo-lactic fermentation
occurs. In this fermentation, malic acid is first
converted to pyruvic acid and then to lactic acid.
FERMENTATION
The reaction is responsible for the rich flavor
developed during the ageing of some wines e.g.
Bordeaux.
Cultures which have been implicated in this

fermentation are Lactobacillus sp and Leuconostoc
sp.
A temperature of 11-16C is best for ageing wines,
high temperature probably functions by accelerating
oxidation.
CLARIFICATION
The wine is allowed to age in a period ranging from
two years to five years, depending on the type of
wine.
At the end of the period some will have cleared
naturally.
For others artificial clarification may be necessary. The
addition of a fining agent is often practiced to help
clarification.
Fining agents react with the tannin, acid, protein or
with some added substance to give heavy quick-
CLARIFICATION
In the process of setting various suspended
materials are adsorbed.
The usual fining agents for wine are gelatin, casein,
tannin, isinglass, egg albumin, and bentonite.
In some countries the removal of metal ions is
accomplished with potassium ferrocyanide known as
blue fining; it removes excess ions of copper, iron,
manganese, and zinc from wines.
PACKAGING
Before packing in bottles the wine from various
sources is sometimes blended and then
pasteurized.
In some wineries, the wine is not pasteurized,

rather it is sterilized by filtration.
In many countries the wine is packaged and

distributed in casks.
CLASSIFICATION OF WINES
CLASSIFICATION OF WINES
PROCESS OF
WINE MAKING
DISTILLED
ALCOHOL (SPIRIT)
BEVERAGES
DISTILLED ALCOHOL
(SPIRIT) BEVERAGES
The distilled alcoholic or spirit beverages are those potable
products whose alcohol contents are increased by
distillation.
In the process of distillation volatile materials emanating

directly from the fermented substrate or after microbial
(especially yeast) metabolism introduce materials which
have a great influence on the nature of beverage.
The character of the beverage is also influenced by such

post-distillation processes as ageing, blending, etc. The
components of spirit beverages which confer specific
aromas on them are known as congeners.
GENERAL PRINCIPLES IN THE

PRODUCTION OF SPIRIT BEVERAGES
In general the following steps are involved in the
preparation of the above beverages. The details differ
according to beverage.
(i) Preparation of the medium: In the grain

beverages (whisky, vodka, gin) the grain starch is
hydrolyzed to sugars with microbial enzymes or with
the enzymes of barley malt.
In all the others no hydrolysis is necessary as sugars

are present in the fermenting substrate as in brandy
(grape sugar) and rum (cane sugar).

(ii) Propagation of yeast inoculum: Large distilleries produce
hundreds of liters of spirits daily for which fermentation broths
many more times in volume are required.
These broths are inoculated with up to 5% (v/v) of thick yeast

broth.
Although yeast is re-used there is still a need for regular inocula. In

general the inocula are made of selected alcohol-tolerant yeast
strains usually Sacch. Cerevisiae grown aerobically with agitation
and in a molasses base.
Progressively larger volumes of culture may be developed before

the desired volume is attained.

(iv) Distillation: Distillation is the separation of more
volatile materials from less volatile ones by a process of
vaporization and condensation.
(v) Maturation: Some of the distilled alcoholic

beverages are aged for some years, often prescribed by
legislation.
(vi) Blending: Before packaging, samples of various

batches of different types of a given beverage are
blended together to develop a particular aroma.
SPIRIT BEVERAGES
Whisky is the alcoholic beverage derived
from the distillation of fermented cereal.
SPIRIT BEVERAGES
Brandy is a
distillate of
fermented fruit juice.
SPIRIT BEVERAGES
Rum is
produced from
cane or sugar
by products
especially
molasses or
cane juice.
SPIRIT BEVERAGES
Cordials are the American name
for what are known as liqueurs in
Europe.
They are obtained by soaking
herbs and other plants in grain
spirits, brandy, or gin or by
distilling these beverages over
the plant parts.
The are usually very sweet, being
required to contain 10% sugar.
Some well-known brand names
of cordials are Drambuie, Crme
de menthe, Triple Sac,
Benedictine, and Anisete.
SPIRIT BEVERAGES
Kai-kai is an
alcoholic beverage
widely drunk in West
Africa. It is produced
by distilling
fermented palmwine.
It is the base for
preparing some of
the better known
brands such as
schnapps.

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INDUSTRIAL

PRODUCTION OF GRAPE WINE

Wine is by common usage defined as a product of the

The production of wine is simpler than that of beer in

PROCESS IN WINE MAKING

The skin contains most of the materials which give

Grapes for sweet wines must have a sugar

The chief sugars in grapes are glucose and

During ripening both the levulose content

Nitrogen is present in the form of amino

PROCESS IN WINE MAKING

Nowadays the must is partially sterilized by the

Other yeasts which have been used for

(a) growth at the relatively high acidity (i.e.,

(b) resistance to high alcohol content

(c) resistance to sulfite.

(a) Temperature: Heat is released during the

(ii) Control of fermentation

(b) Yeast Nutrition: Yeasts normally ferment the

(c) Oxygen: As with beer, oxygen is required in

(iii) Flavor development: Although some flavor

The flavor of wine has been elucidated with gas

Autolysates from yeasts also have a special influence

PROCESS IN WINE MAKING

AGEING AND STORAGE

The wine is allowed to stand until a major portion

During ageing desirable changes occur in the wine.

(a) Slow oxidation, since oxygen can only diffuse

Cultures which have been implicated in this

PROCESS IN WINE MAKING

PROCESS IN WINE MAKING

In some wineries, the wine is not pasteurized,

In many countries the wine is packaged and

In the process of distillation volatile materials emanating

The character of the beverage is also influenced by such

GENERAL PRINCIPLES IN THE

(i) Preparation of the medium: In the grain

In all the others no hydrolysis is necessary as sugars

GENERAL PRINCIPLES IN THE

These broths are inoculated with up to 5% (v/v) of thick yeast

Although yeast is re-used there is still a need for regular inocula. In

Progressively larger volumes of culture may be developed before

GENERAL PRINCIPLES IN THE

(v) Maturation: Some of the distilled alcoholic

(vi) Blending: Before packaging, samples of various

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