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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 18 th 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.

1950’s

- 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.

to 1960’s

Alanine added to fruit juice to improve taste Aspartate

– added to fruit juice to improve taste Aspartate Cysteine added to bread and fruit juice

Cysteine

added to bread and fruit juice to improve flavor

Glutamate enhance flavor (MSG)

Glycine

Histidine and trypsin prevents food rancidity

- enhance flavor of sweetened food

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 14-23ºC for 5-7 days.

g. The fermentation product is pumped into a large tank. Lager beer is stored at 1°C and ale is stored at 4-8ºC 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

Grapes

crushed

crushed

SO 2

SO 2 must or must or
SO 2 must or must or

must or

SO 2 must or must or

must or

juice

juice

Juice extracted juice sits in contact with skin (6-24 hr)

Fermentation vat 3 weeks Pulp is not removed

(6-24 hr) Fermentation vat 3 weeks Pulp is not removed Press Discard pomace Fermentation vat 10-15

(6-24 hr) Fermentation vat 3 weeks Pulp is not removed Press Discard pomace Fermentation vat 10-15

Press

Discard

pomace

Fermentation vat 10-15 days

SO 2

Press Discard pomace Fermentation vat 10-15 days SO 2 Aging in 5 months clarifying agent casein
Press Discard pomace Fermentation vat 10-15 days SO 2 Aging in 5 months clarifying agent casein

Aging in 5 months

clarifying

agent

casein

tannin

Filtration
Filtration

Racking : separate wine from sediment & yeast

Filtration Racking : separate wine from sediment & yeast Bottling WHITE WINE SO 2 Press Discard

Bottling

WHITE WINE

SO 2

wine from sediment & yeast Bottling WHITE WINE SO 2 Press Discard pomace Aging in barrels

Press

Discard

pomace

Aging in barrels

WHITE WINE SO 2 Press Discard pomace Aging in barrels Racking : separate wine from sediment
WHITE WINE SO 2 Press Discard pomace Aging in barrels Racking : separate wine from sediment

Racking : separate wine from sediment & yeast

Transfer to clean barrels 3x/year

clarifying agent casein Settling tank tannin
clarifying
agent
casein
Settling tank
tannin
Filtration
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.

confirmed as a pure line that gives same plants each time. PLANT 2 Plant with poor

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

confirmed as a pure line that gives same plants each time. CROSS POLLINATION F 1 HYBRIDS

CROSS POLLINATION

line that gives same plants each time. CROSS POLLINATION F 1 HYBRIDS WITH COMBINED DESIRABLE TRAIT

F 1 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 1920’s 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 1960’s 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.

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.

Live attenuated vaccines

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

Not sensitive

Yes

Yes

Duration of immunity

Many years

Often less

Safety: Reversion to virulence

Rarely

No

Side effects

Low levels 1-2 for every 10 6

No