PRODUCTS OF TRADITIONAL BIOTECHNOLOGY

No. 2

TRADITIONAL BIOTECH CUISINES

Fermented Food Products
Bread, yogurt, cheese, wine, and beer are produced by fermentation.
Fermentation involves a microbial process where enzymes produced by
microorganisms catalyze the transformation of organic compounds. Fermentation was
derived from the Latin word fervere which means to boil. It was observed that the
addition of yeast to fruit juices or cereal grains during bread-making produce bubbling
from carbon dioxide production. The trapped carbon dioxide causes the bread to rise.
In beer and wine making, yeast converts sugar to alcohol. It is believed that fermented
food products were produced initially by accident.
4000 BC The Chinese already use lactic acid producing bacteria for making yogurt, use
molds for making cheese, and acetic acid bacteria for making wine vinegar. To
date soy sauce, fermented rice, vegetables and fruits, are still being made with
salt or brine producing our fermented olives, pickles, and sauerkraut.
Between 5000 to 9000 BC
Milk from animals has been used for cheese making. Now, the same
technology is used for making cheese, Cheese makers inoculate the milk with lactic
acid bacteria and add enzymes (chymosin) also called rennet, to curdle the casein.
Camembert and Roquefort cheese (French cheeses) using molds were created by
accident in the late 18th century. So these are traditional biotech cuisines.
Camambert cheese was made when the curd was inoculated with a mold
Penicillium camamberti and was commercially produced in the 1800. Roquefort
cheese is made when the curd was contaminated with a mold Penecillium glaucum
roqueforti.
1500 BC
It is believed that fermented dough has been discovered by accident when some
dough was not baked immediately, underwent spontaneous fermentation and when
baked, produced a lighter, expanded, and more palatable bread. Later, most
probably, they also discovered that uncooked fermented dough could be used to
ferment a new batch of dough the beginning of starter dough.

Fermented Beverage
Beer making began as early as 6000 and 5000 BC. This process made use of
dough from barley that is partially baked before it is allowed to undergo
fermentation in water to produce acid beer. The acid beer is put in a jar and stored.
1680 - Dutch biologist and microscopist Anton van Leewenhoek examined
samples of fermenting beer through and observed yeasts under the
microscope
1800 Brewers are already producing alcohol on a large scale in 1800.
Brewers had accumulated enough knowledge to use pure yeast
cultures in the fermentation process
1837 The connection was made between yeast cell activity (observed by
Leewenhoek) and alcohol fermentation.
1876 Louis Pasteur, a French chemist, established that yeast and other
microbes are linked to fermentation and described that yeast
convert sugar into ethanol and carbon dioxide.
1911 Brewers already measure the amount of acid during mashing to better
control quality of beer
It is also believed that wine was first made accidentally when juices from grapes
fermented naturally in the presence of yeast, producing alcohol. When wine is
allowed to sit in shallow barrels, it is oxidized to vinegar by the action of
bacterium Acetobacter sp.
Other fermentation products were later produced in the 1900-1940 for different
uses such as glycerol, acetone, butanol, lactic acid, and citric acid. Yeast
biomass was also a product of fermentation. Glycerol, acetone, and butanol
were used during World War I for explosives. Yeast biomass was also used
during the war as source of single cell protein.
1950s to 1960s - Microbial production of antibiotics and amino acids
occurred in response to the need for antibacterial cure during World
War II. The same technology is still useful for antibiotic production.
Alanine added to fruit juice to improve taste
Aspartate
Cysteine
added to bread and fruit juice to improve flavor
Glutamate enhance flavor (MSG)
Glycine - enhance flavor of sweetened food
Histidine and trypsin prevents food rancidity

Lysine added to bread

Methionine added to soybean

BEER BREWING
a. Malt is prepared from barley placed in vessels and buried until barley is

germinated. Germinated barley (which is called malt) which contains

starch, sugars, coloring, aromatic compounds and natural enzymes that
digest starch grains into sugar.
b. Mash is produced by combining malt and grains (barley) and cooked. At
this point, the enzyme from malt cause the digestion of starches to sugar.
c. The aqueous extract (now called wort) is obtained from the fermented mash
and filtration is performed.
d. Hops (herb from female flower of hop plant) is added for flavor to the wort. Hops
are also believed to have antimicrobial activity.
e. Wort is boiled for several hours in large copper kettles, filtered, cooled, and
transferred to fermentation vessel.
f. Yeast is added for the fermentation of sugars which produce alcohol.
Bottom yeast (Saccharomyces carlsbergensis) is used to produce
lager beer. Fermentation is at 6-12C for 8-14 days.
Top yeast (Saccharomyces cerevisiae) is used to produce ales.
Fermentation is at 14-23C for 5-7 days.
g. The fermentation product is pumped into a large tank. Lager beer is stored
at 1C and ale is stored at 4-8C prior to filtration and bottling.

EARLY AGRICULTURAL BIOTECHNOLOGY

Paleolithic people lived in mobile camps and survived by hunting wild
animals and collecting wild plants.
About 10,000 years ago, people settled and began domesticating plants and
animals and unconsciously started the first forms of genetic selection.
Early farmers (from Iran, Iraq, Turkey, Syria, Lebanon, Israel) are believed to
have cultivated wheat, barley, and rye. Sheep and goats provided milk,
cheese, butter, and meat.
About 7,000 years ago, Africa (not yet a desert then) had flocks of sheeps,
goats, and cattle.
Evidence suggests that agriculture developed independently in several areas
of the world. People collected seeds of wild plants for cultivation and
domestication of wild animals.
Farmers collected seeds of plants with most desirable traits and set them
aside for planting the next year. They bred only the most prized animals,
thus illustrating the practice of artificial selection that produced new varieties
of plants.

Therefore, new varieties retained the desirable traits found in the wild
species but were modified in other ways, a form of ancient genetic
manipulation.
Selective breeding for improving livestock was also performed in the same
manner.
Plant collecting expeditions were common and collectors traversed the
globe. Plants include beans, peas, maize, lentils, oats, rye, and wheat.
Nikolai I. Vavilov (1887-1943). Russian plant geneticist and agronomist,
initiated comprehensive research and breeding program and was first to
introduce the concept of crop genetic resource management. He was,
however, arrested in 1940 on charges of espionage and died in prison from
malnutrition in 1943.

WINE MAKING
Grapes
crushed
SO2

Grapes
crushed
must or
juice

must or
juice

Juice extracted juice sits in

contact with skin (6-24 hr)

Press

Fermentation vat 3 weeks

Pulp is not removed

Press

Discard
pomace

Fermentation vat 10-15 days

SO2

clarifying
agent
casein
tannin

Aging in barrels
Racking : separate wine
from sediment & yeast

SO2
Aging in 5 months
Racking : separate wine
from sediment & yeast

Filtration

Bottling

WHITE WINE

Discard
pomace

Transfer to clean barrels 3x/year

clarifying
agent
casein
tannin

Settling tank

Filtration

Bottling
Age in bottles
6 months or

RED WINE

 Trofim Lysenko (1898-1976) a leader in Soviet Science under Stalen, rejected

the principles of Mendelian genetics in favor of the Lamarckism.
(Lamarckism is the theory that an organism can acquire physical traits in
response to its environment and pass it to the offspring). Therefore, this
resulted in the banning of selective breeding and experimentation on
scientific inquiries.
In New York (1948), Georgia and Washington State (1949), centers for
germplasm preservation, study, and distribution were established. USDA
established a national gene bank, the National Seed Storage Laboratory, Fort
Collins, Colorado.
Traditional plant breeding is based on the principles of Mendelian genetics.
Selection of superior genotypes of a particular crop depended on subjective
decisions made by the breeder.

CONVENTIONAL PLANT BREEDING

Since the practice of agriculture about 8,000 years ago, man has been trying to
alter the genetic make up of plants. They select and decide desirable characteristics
and use such selected plants in conventional breeding procedures. Among the
desired characteristics chosen are high yields, disease resistance, fast growth, and
better quality fruits, or seeds. Therefore this process depends a lot on the proper
recognition of desirable traits. How do the plants acquire more desirable traits in
nature? This occasionally arise through a process of mutation which is a natural
process. In nature, however, these mutations that result in desirable plant
characteristics, is a very slow process.

HYBRID SEED TECHNOLOGY

This involves the crossing of pure lines to produce F1 hybrids with predictable
characteristics. For example :

PLANT 1
Plant with good growth but
poor color is self pollinated until
confirmed as a pure line that
gives same plants each time.

PLANT 2
Plant with poor growth but good is self
pollinated until confirmed as a pure
line that gives same plants each time.

CROSS POLLINATION
F1 HYBRIDS WITH
COMBINED DESIRABLE
TRAIT OF GOOD GROWTH
AND GOOD COLOR

 A completely pure line can sometimes take seven to eight years to

achieve
Seeds are more expensive since it took a long time to produce and
is expensive to maintain
Cross pollination usually has to be done by hand to ensure self
pollination, therefore labor intensive
Farmer may need to buy hybrids each year since performance of
F1 may not be maintained.
Today, nearly all corn and 50% of all rice are hybrids.
MUTATION BREEDING
In the late 1920s researchers discovered that the number of variations or
mutations in plants can be increased by exposing plants to Xray, UV, and
chemicals. This has been used with wheat, barley, rice, potatoes, soybeans, and
onions. There is FAO mutant Variety Database maybe accessed which will show
variants (http://ww-mvd.iaea.org/MVD/default.htm) produced by mutation in
specific crops.

LIMITATIONS
1. Breeding can only be done between two plants that can sexually mate.
2. In crossing, many traits are transferred along with traits of interest.
Including those that are undesirable.

CONVENTIONAL PROCEDURE TO ADDRESS PESTS AND

DISEASES OF PLANTS AND ANIMALS
How plant and animal diseases are recognized
Visual examination for symptoms (but sometimes symptoms do not appear till
the enough damage has been done)
Microscopic observation
Biochemical tests
Culture and growth of microorganisms
How the disease were addressed and are still currently addressed
Plant pest
Bio insecticides
Chemical insecticides
Antimicrobial formulations

Animal disease
Use of antibiotics
Vaccination

Vaccination
Vaccination has been one of the most useful and successful discoveries as early
as the 1960s in human and veterinary medicine to address viral and bacterial diseases.
Conventional vaccines include live, attenuated, and killed vaccines.
Inactivated whole vaccines are made from diseases causing organisms or
pathogens. The infectivity of pathogens is destroyed with formalin while retaining its
ability to elicit an immune response in the animal host.
Live attenuated vaccines are live but weakened derivatives of pathogenic
organisms. Most of this type of vaccine have been derived by passage in culture until
they have lost virulence or ability to cause disease.
Major Points on Live vs killed vaccines

BASIS OF COMPARISON

LIVE-ATTENUATED

KILLED

Production

Relatively simple

More complex

How it is used

Injection

Injection

Dose

Low, often single

High, multiple

Heat sensitivity
Need to refrigerate

Sensitive
Yes

Not sensitive
Yes

Duration of immunity

Many years

Often less

Safety: Reversion to
virulence

Rarely

No

Low levels
1-2 for every 106

No

Side effects