ALCOHOLIC

BEVERAGES
 INTRODUCTION

• All alcoholic beverages involve the action of fungi.

• Most involve the genus Saccharomyces. These yeasts covert
six carbon sugars such as glucose to ethanol and carbon
dioxide and live under anaerobic conditions. Yeasts can
tolerate fairly high concentrations of alcohol (up to about 14-
18%) in the medium.
• Alcoholic beverages are known from virtually all cultures.
These beverages are major social problems, but also sources
of nutrition.

• Plants often store nutrition as starch. Yeasts cannot use starch.

The starch is broken down by enzymes in the plant into
sugars. The sugars are then converted by the yeasts into
ethanol and carbon dioxide.
• Alcohol is a lipid and moves freely across membranes in the
stomach.
• Alcohol is broken down in humans, but also affects the
neurons and is a non-selective central nervous system
depressant.
WHAT IS FERMENTATION?
A metabolic process that converts sugar
to acids and gases, or alcohol. It occurs
in yeast and bacteria, and also in oxygen-
starved muscle cells, as in the case of
lactic acid fermentation.

ZYMOLOGY – or ZYMURGY – is an applied science

which studies the biochemical process of
fermentation and its practical uses.
HISTORY OF FERMENTATION
ANCIENT TIMES
7000 BC Beer and wine Assyria, Caucasia, Mesopotamia, Sumer
6000 BC Wine making Georgia
5000 BC Wine jars (artifacts) Iran
3000 BC Beer and fermented milk products Babylon
2600 BC Bread Egypt
1000 BC Soy sauce & miso China
600 BC Cheese Asia
500 BC Preservation of fish & meat
100 BC Bread Ancient Rome
HISTORY OF FERMENTATION
MODERN TIMES
Vinegar from fruit pulp (remains of grapes, olives, or other fruit after pressing
1700s
for juice or oil); gallic acid
1800s Yeast induced fermentation Germany
o Bacteria produce lactic acid which preserves food.
o Pasteurization – heat treatment to prevent unwanted fermentation. France
1850s
o Yeast + Grape juice = wine; the beginning of the science of food - Louis Pasteur
fermentation.
LATE
Composting
1800s
1900s Aseptic fermentation (no unwanted organisms)
HISTORY OF FERMENTATION
NEW FERMENTATION
Industrial Production of Acetone, Butanol, Butanediol – Substrates For
1900 - 1920S United Kingdom, Germany , Russia
Rubber Production
Development of biotechnological methods of organic acid production – citric
1930S
& gluconic acid
1940S Industrial production of organic acids
Industrial penicillin production, the first bacterial antibiotic; first large scale
1945 - 1950 United States
production of pharmaceuticals
1950S -
Steroid transformations by fungal spores
1960S
Commercial production of amino acids using fermentation; production of
1960S Japan
MSG, a flavor enhancer
1960 - 1980S Mycotoxins, treatment and reuse of wastes
Enzymes produced by microbes for use in detergents, grain processing,
1970S World-wide
sugar production and fruit juice clarification
1980S Genetic Engineering techniques – Insulin production United Kingdom
 USEFUL PRODUCTS OF
FERMENTATION

LACTIC ACID ETHANOL

PRODUCTS OF FERMENTATION

Beer Wine
PRODUCTS OF FERMENTATION

Mead Cider
PRODUCTS OF FERMENTATION

Yogurt Cheese
THE GLYCOLYTIC PATHWAY

ETHANOL
GLUCOSE PYRUVATE or
LACTIC ACID

GLYCOLYSIS
THE GLYCOLYTIC PATHWAY

ETHANOL
GLUCOSE PYRUVATE or
LACTIC ACID

C6H12O6 + 2 NAD+ + 2 ADP + 2 Pi → 2 CH COCO

3 2
− + 2 NADH + 2 ATP + 2 H2O + 2H+

glucose pyruvate
 GLYCOLYSIS

GLUCOSE
A six-carbon molecule that is a
digestive product of
carbohydrates and plays a major
role in the metabolism of living
C6H12O6
cells.
 GLYCOLYSIS
Step 1 - hexokinase
Phosphorylation
A reaction that transfers a
phosphate group from one
molecule to another.
 GLYCOLYSIS
Step 1 - hexokinase
hexokinase
The glycolytic enzyme responsible
for catalyzing the reaction that
transfers a phosphate group from
a molecule of ATP to a 6-
membered glucose ring.
 GLYCOLYSIS
Step 2 – phosphoglucose
isomerase the conversion of
glucose-6-phosphate to
fructose-6-phosphate
 GLYCOLYSIS
Step 2 – phosphoglucose
isomerase This reaction occurs with
the help of the enzyme
phosphoglucose
isomerase (PI).
 GLYCOLYSIS

Step 3 - phosphofructokinase
fructose-6-phosphate is
converted to fructose-
1,6-bisphosphate (FBP)
using atp.
 GLYCOLYSIS

Step 3 - phosphofructokinase
The enzyme that
catalyzes this reaction is
phosphofructokinase
(PFK)
 GLYCOLYSI
S
Step 4 - aldolase
The final step of the first
stage of glycolysis utilizes
the enzyme aldolase, which
catalyzes the cleavage of
FBP to yield two 3-carbon
molecules.
 GLYCOLYSIS
Step 5 - Glyceraldehyde-3-phosphate Dehydrogenase

two main events take place:

1) glyceraldehyde-3-phosphate is
oxidized by the coenzyme
nicotinamide adenine dinucleotide
(NAD)

2) the molecule is phosphorylated by

the addition of a free phosphate
group.
 GLYCOLYSIS
Step 5 - Glyceraldehyde-3-phosphate Dehydrogenase

The enzyme that catalyzes

this reaction is
glyceraldehyde-3
phosphate dehydrogenase
(GAPDH).
 GLYCOLYSIS

Step 6 - Phosphoglycerate Kinase

1,3 bisphoglycerate is
converted to 3-
phosphoglycerate by the
enzyme phosphoglycerate
kinase (PGK).
 This step involves a simple
GLYCOLYSIS rearrangement of the
position of the phosphate
group on the 3
Step 7 - Phosphoglycerate MUTASE phosphoglycerate molecule,
making it 2
phosphoglycerate. The
molecule responsible for
catalyzing this reaction is
called phosphoglycerate
mutase (PGM).
 The eighth step involves the
GLYCOLYSIS conversion of 2
phosphoglycerate to
phosphoenolpyruvate (PEP).
Step 8 - enolase The reaction is catalyzed by
the enzyme enolase.
Enolase works by removing
a water group, or
dehydrating the 2
phosphoglycerate.
 GLYCOLYSIS
STEP 9 – PYRUVATE KINASE
The final step of glycolysis
converts phosphoenolpyruvate
into pyruvate with the help of
the enzyme pyruvate kinase.
THE GLYCOLYTIC PATHWAY

ETHANOL
GLUCOSE PYRUVATE or
LACTIC ACID

FERMENTATI
ON
 ANAEROBIC ALCOHOLIC
FERMENTATION
In alcoholic fermentation, the pyruvate is converted into carbon
dioxide gas and ethyl alcohol (ethanol).

C6H12O6 → 2 C2H5OH + 2 CO2

2 CH3COCO2− + 2 NADH → 2 NAD+ + 2CO2 + 2 C2H5OH
 ANAEROBIC HOMOLACTIC
FERMENTATION

Homolactic fermentation (producing only lactic acid) is the simplest

type of fermentation. The pyruvate from glycolysis undergoes a
simple redox reaction, forming lactic acid.

C6H12O6 → 2 CH3CHOHCOOH
2 CH3COCO2− + 2 NADH → 2NAD+ + 2 CH3CHOHCOOH
WINE MAKING
 WINES

• Wine is fermented fruit juice. The most important fruit is Vitis

vinifera (Vitaceae), but any fruit can be used.
• Yeasts occur on the skins of most fruits and if the fruits are
mashed, the sugar-containing juices begin to ferment.
GRAPE FLOWERS AND FRUITS, VITIS VINIFERA, VITACEAE
YEAST ON THE OUTSIDE OF GRAPES B. Lehane, Power of Plants, McGraw Hill. New York.
1977
• Winemaking probably began as one of the earliest of human
enterprises (8000-3000 B.C.).
• The wine grape was domesticated by at least 4000 B.C. Wine
was used for Egyptian worship ceremonies.
• Wine only became a popular beverage about 2000-1000 B.C.
in Greece.
• About 600 B.C., wine growing reached France.
 GRAPE MOTIF IN AN
EGYPTIAN TOMB

B. Lehane, Power of Plants, McGraw Hill. New York.

1977
VINEYARDS NEAR
LAUSANNE,
SWITZERLAND
• Many good quality wines are still made in the same way they
have been made for centuries. However, most wines are now
made by sophisticated highly controlled processes.
• Grapes are crushed (still by foot or with crude presses in many
parts of the world).
• The juice is then often treated with sulfur dioxide to kill native
yeasts.
 1 - HARVESTING

Hand
Picking
Harvester
 2 - CRUSHING

Sorting Destemming
 3 - PRESSING

Pressing Pressing
NEWER STYLE EQUIPMENT FOR PRESSING GRAPES
GRAPE POMACE AFTER PRESSING
4 – MUST ADJUSTMENTS
 5 - FERMENTATION

Malolactic
Lees Fermentation
• If white wine is to be made, the juice is put into fermentation
tanks and the peels and stems re-pressed.
• If red wine is to be made, the skins are added.
• Special strains of yeasts are added to the liquid from the
grapes. These are often highly guarded trade secrets and
proprietary substances.
• After 8-10 days, the liquid is drawn off and allowed to ferment
for another 20 days to one month. Sediment forms.
WINE FERMENTATION
TANKS
 6 - AGING

Wine Aging in Oak

Barrels
AGING OF WINE IN BARRELS
STAINLESS STEEL AGING TANKS FOR WINE
7 - BOTTLING

Bottling
AGING OF CHAMPAGNE IN A NEW YORK WINERY
• Apples and pears are used to make cider and perry
respectively.
• To make fortified wines, ethanol or distilled beverages such as
cognac are added.
• As many as 40 million metric tons of grapes are produced per
year.
• The major wine making countries are the former USSR,
France, Italy, USA, Spain, Germany, Australia, Argentina, Chile
and South Africa.
• Consumption of wine in France is about 30 gal per person per
year and about 2 in the U.S.
 BEER, ALE AND STOUT
• Beer making goes back at least 6000 years. The Sumerians had records about brewing
of beer.
• Early brewing is intimately linked with bread making. This is one way of making the
grains more edible.
• Although the Egyptians used wine for religious ceremonies, they commonly drank beer.
Barley fields in California
and Germany
• Yeasts were developed and saved to give better qualities of
beer. Today beer making is also a quite sophisticated process.
• The three main ingredients are barley malt, hops and water.
• As much as 7.5 X 109 liters of beer are made per year (2 X 109
gallons).
 MALT

• Malt is sprouted grain that has been dried.

• Barley (Hordeum vulgare, Poaceae or Gramineae), is
preferred for several reasons. The husks stay on the kernels
and add some of the flavor. Also, barley contains higher
concentrations of the enzymes needed for converting starches
into sugars.
• The grain is steeped in water and allowed to sit where it is
warm until germination begins. The grain synthesizes
hydrolytic enzymes that convert starch to sugar. The cell walls
of the endosperm break down. The mixture is then dried at
130 to 200 C.
 THE BREWING PROCESS
• The process is now highly mechanized.
• The malt is extracted at 68-73 °C for 2-6 hours, a process
called mashing. During mashing, the enzymes diffuse into the
solution and break down the starch in the malt and the
adjuncts. Proteins are degraded into amino acids.
 THE MALTED GRAIN IS
DRIED, MILLED,
MASHED, AND STRAINED.
ADJUNCTS ARE ADDED
AT THIS STAGE.

Anheuser Busch Brewing Co., St. Louis

 ADJUNCTS

• Adjuncts are unmalted grains that are added. Barley, rice,

wheat, or corn grits are often used.
• In other countries, potatoes or cassava may be used.
• Adjuncts are commonly used in the U.S., but usually not in
Europe. They are less expensive than barley malt. Corn and
rice are both precooked before being added.
THE WORT IS MADE BY
BOILING THE
EXTRACTED MALT
MIXTURE WITH HOPS.

Anheuser Busch Brewing Co., St. Louis

 HOPS (HUMULUS LUPULUS, CANNABACEAE)

• Hops (Humulus lupulus, Cannabaceae) were only added to

beer after 700 A.D.
• The English only started adding them in the 1500's.
Sometimes other plants were added for flavoring before that
time. Hops not only add flavor but help to coagulate and
precipitate proteins.
• The female inflorescences are used. They contain a glandular
exudate with the flavor properties.
HOPS, HUMULUS LUPULUS, CANNABACEAE, FRUITS
HOPS IN CULTIVATION IN
KENT, ENGLAND
• Hops are then added and the liquid boiled.
• This boiling step does several things.
• The yeasts present are killed.
• Compounds in the hops are converted to bitter tasting
compounds.
• The resulting liquid, called wort, contains sugars, some
starches, proteins and amino acids, among other
components.
• The spent malt and adjuncts often are used for cattle
feed.
• In the process of making the wort, enzymes in the mixture are
denatured.
• Compounds in the hops are converted into the compounds that
give beer its bitter taste. The hops are then removed.
• The liquid is pumped to fermentation tanks and yeast is added.
• Saccharomyces cerevisiae is a top fermenting yeast and S.
uvarum is a bottom fermenting yeast. The latter is used to
make ales and lager beers.
 FERMENTATION AND
AGING OF BEER

Anheuser Busch Brewing Co., St. Louis

• Carbon dioxide is captured and is a major byproduct of
brewing.
• The mixture is allowed to ferment for 7-12 days at cool
temperatures.
• The resultant liquid is transferred and allowed to age for 2-3
weeks.
• Proteins precipitate out and some other chemical modifications
take place.
• In the U.S., most of the beers are pasteurized or micropore
filtered to remove yeasts. The beer is usually carbonated.
 SAKE
• Sake is made from rice and is a traditional beverage in Japan.
It is a fermented grain beverage.
• Aspergillus oryzae is used to convert the starch to sugar and
then yeast added. Both are involved in the fermentation
process.
• The mixture is fermented for about 25 days.
• The alcohol content is about 18-19%. The beverage is allowed
to mature for about 40 days. It is consumed before 1 year.
 PULQUE

• Pulque is made by fermenting the juice of Agave species,

especially A. americana. Pulque made from Agave tequilana is
distilled to produce tequila.
MAGUEY, AGAVE AMERICANA, AGAVACEAE

http://commons.wikimedia.org/wiki/Image:Agave_americana.jpg
COLLECTING THE AGUA
MIEL WITH AN OCOTE
TRANSFERRING THE AQUA MIEL FOR TRANSPORT
FILTERING THE AQUA MIEL PRIOR TO FERMENTATION
 CHICHA

• Chicha is made by chewing cassava or corn and spitting the

materials into a tank or container. The mixture is then allowed
to ferment. The saliva serves as a source of amylase to break
down starches. This beverage is found in many areas of
western South America.
“MASH” AND FERMENTATION OF
CHICHA IN PERU
Courtesy Dr. Memory Elvin-Lewis
DR. MEMORY ELVIN-
LEWIS WITH A BOWL
OF CHICHA IN PERU

Courtesy Dr. Memory Elvin-Lewis

 OTHER ALCOHOLIC BEVERAGES

• Kvass is made from fermented bread, barley or rye. It is

sometimes peppermint flavored and is popular in the former
Soviet Union.
• Sorgo is made from fermented sorghum in Africa.
• Pombe beer is made from bananas in Uganda and eastern
Africa.
 DISTILLATION

• Distillation involves converting components to the vapor phase

and then condensing them.
• The Arabs probably discovered this process. They distilled
perfume components and other mixtures of fermented
materials.
• Many of these substances were prepared for medicinals.
 Brandies

• Brandies are distilled wines.

• The most famous come from France. Cognac is one type.
• The fermented juices of many other fruits are also distilled.
• Liquers differ from brandy in that various flavoring agents are
added. They have sugar and syrups added.
ALEMBICS FOR DISTILLING
WINE TO MAKE BRANDY.
• By the 15th century, the English and Scots had begun distilling
barley beer and, in the 16th century, cognac was prepared in
France. Whiskeys were made in a similar way.
• Once distilled, the mixture is diluted back to about 50% alcohol
in general. Whiskeys are aged in (usually oak) barrels.
• The inside of these barrels is often charred. The barrel
provides some of the flavoring materials.
• Scotch whiskeys have a characteristic taste because of the
smoking process (over peat) used to dry the malt.
• Bourbon whiskey is made from corn as the primary grain (Zea
mays, Poaceae) and was developed by the early Scottish
(many of whom are called Scots-Irish) immigrants in
Pennsylvania.
• These whiskies are aged for at least two years in new, charred
oak barrels.
AGING OF WHISKEY IN CHARRED WHITE OAK BARRELS
 OTHER DISTILLED BEVERAGES

• Gin and vodka are distilled to a high percentage of alcohol and

in the case of gin, Juniperus communis, Cupressaceae,
"berries" are added.
• These beverages can be distilled from almost any fermented
mixture including potatoes, grains, etc.
• Rum is made from fermented molasses or sugar cane juice.
 TEQUILA AND MESCAL

• Tequila and mescal are distilled from pulque made from

various Agave species.
CULTIVATION OF AGAVE TEQUILANA IN JALISCO, MEXICO
 FIELD OF AGAVE AND
“HEARTS” REMOVED
FROM MATURE PLANTS.
 Courtesy Dr. Beryl Simpson

HEARTS OF AGAVE ON
WAY TO DISTILLERY.
STEAMED HEARTS READY TO BE FERMENTED AND DISTILLED.

Courtesy Dr. Beryl Simpson