ABT 344 FOOD BIOTECHNOLOGY 1+1

THEORY
Biotechnology-scope-principles-genetic engineering-gene concept-DNA and RNA
genetic code-organization of genetic material in virus, prokaryotes and eukaryotesgenetic improvement of microorganism-mutation-recombinant DNA technologyIndustrial Fermentations-Fermentations of alcohol, Citric acid, acetic acid and lactic acidAntibiotic fermentation-fermentation for enzymes, Vitamins and growth hormones, food
fermentations-dahi (Curd)cheese-yoghurt-sauerkraut pickles-bread-idli batter-microbes as
food and feed-single cell protein-biomass production of plants. In vitro propagation of
ornamental plants-lilies, orchids, roses, asparagus and chrysanthemum-mass production
of food crops through tissue culture methods. Biotechnology of food-Development of
Fruits and Vegetables into drugs, Vaccines and antibiotics.
PRACTICAL
Studies on sterilization technique-Media for tissue culture-development of mutantsrecombinant DNA technology-production-fermented food-alcohol-antibiotics-organic
acid-Enzymes-In vitro propagation of rice-micropropagation of ornamental plants.
LECTURE SCHEDULE
1. Biotechnology-definition-scope in different areas of biotechnology viz., microbial
biotechnology,

Plant

biotechnology

and

animal

biotechnology-principles

of

biotechnology
2. Genetic engineering-gene concept-DNA-RNA-fine structures of these genetic
materials-genetic code

3. Organisation of genetic materials in virus, prokaryotes and eukaryotes

4. Genetic improvement of microorganisms (bacteria and fungi) by mutation and
recombinant DNA technology
5. Industrial fermentations-alcohol fermentation by yeasts and bacteria
6. Citric acid fermentation, acetic acid fermentation and lactic acid fermentation
7. Antibiotic fermentations-penicillin, streptomycin and tetracyclines
8. Fermentation enzymes-amylases, proteases and cellulases
9. Mid semester examination
10. Fermentation of vitamins, growth hormones, dahi(curd) cheese, Yogurt, sauerkraut
and pickles
11. Bread fermentation and idli batter fermentation
12. Biomass production by microbes-bacteria, fungi and algae-biocides for the control of
pests and diseases
13. Microbes as food and feed-acceptability of single cell protein
14. Biomass production of plants-in vitro propagation of ornamental plants viz., lilies,
orchids, roses, asparagus and chrysanthemum
15. Mass production of food crops through tissue culture-methods viz., Callus induction
and regeneration of plant-rice
16. Biotechnology of food-definition and scope
17. Developing fruits and vegetable as carriers of drugs, vaccine and antibiotics

PRACTICAL SCHEDULE
1. Preparation of tissue culture media
2. Development of bacterial mutants by Physical method
3. Development of bacterial mutants by chemical method
4. Development of bacterial transformant
5. Production of single cell protein (Yeast, spirulina)
6. Production of alcohol from molasses by yeast
7. Production of pickles-sauerkraut
8. Production of alcohol from molasses by bacteria
9. Production of penicillin
10. Production of enzyme cellulases by fungi
11. Production of vinegar
12. Production of wine in laboratory condition
13. Production of cheese in laboratory condition
14. Micropropagation of Rose and Chrysanthemum
15. Studies on callus induction and regeneration of rice
16. Studies on callus induction and regeneration of banana
17. Practical examination

REFERENCES
1. Brown, C.M., I.Campbell and F.G.Prest, 1988. Introduction to biotechnology, ELBS.
Black well Scientific Publication, Oxford.
2. Chakraverty, A., 1989. Biotechnology and Other Alternative Technologies, Oxford and
IBH Publishing Co., Pvt., Ltd.,
3. Kalyan Kumar, De., 1992. Plant tissue culture. New Central Book Agency, Calcutta-9.
4. Ramachandran, S., 1989. Biotechnology in Agriculture. Oxford and IBH publishing
Co., Pvt., Ltd., New Delhi.
5. Pepplor, J.H., 1967. Microbial technology. Revinhold Publishing Corporation, New
York.
6. Purchit, SS and S.K.Mathur, 1992. Fundamentals of biotechnology. Agro. Botanical
Publishers, India.
7. Rama, S.V.S., 1986. Recent trends in Biotechnology and Biosciences. Society of
biosciences, Iragati Press, Muzaffarnagar.

Course Teacher

G.Gopalaswamy, professor (Biotech)

Course Associate

P.Maheshwari

Lecture schedule 1: Biotechnology-scope-principles

Sl.no.
1.

List of Contents
Biotechnology principles and scope

2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.

Areas of biotechnology
Microbial biotechnology
Some bacteria of biotechnological importance among 19 bacterial groups
Plant biotechnology
Insect protection and advantages
Disease protection
Advantages of biotechnology over irraditional breeding
Animal biotechnology
Other uses of biotechnology in animal production
Principles of biotechnology
Benefits and risks of biotech foods

FOOD BIOTECHNOLOGY (1+1)

1.

BIOTECHNOLOGY PRINCIPLES AND SCOPE

The term biotechnology usually applies to the use of recombinant DNA technology.
However, it has a broader definition, covering any technique that involves the use of
biological species and biomass, or their derivatives for the generation of beneficial
products. Though the term biotechnology was first coined by Karl Erkey in 1917, the
comprehensive definition as defined by the US congress in 1984, indeed states that it is
any technique that uses living organisms, to make or modify products, to improve plants
or animals, or to develop microorganisms for specific uses.
The term biotechnology is now refers to the use of recombinant DNA and in vitro
biological techniques in three major areas viz.,
1. As powerful tools in classic breeding
2. As means for generating transgenic plants, animals, and other organisms, and
3 As a means of integrating microorganisms into various agricultural production systems.
We are looking at a doubling of the population in the next 40 years. Were looking at a
need for food production increase of 250 per cent. At the same time, were looking at

dwindling resources for that food production. So, clearly, biotechnology with its ability to
improve yield, quality and nutritional value will help us in feeding and tomorrows
population.
-Terry Nedley, J.D.
- Animal and Plant Health Inspection Service, USDA.

AREAS OF BIOTECHNOLOGY
1. Agricultural biotechnology
a. Soil
b. Plant
c. Animal
d. Microbes Bioinoculants
2. Industrial biotechnology
3. Pharmaceutical biotechnology
4. Environmental biotechnology
5. Food biotechnology
6. Marine biotechnology
Microbial biotechnology
Microbes play an important role. Mostly they are ubiquitous and highly adaptive
to varied environments including extreme environments. Biological Nitrogen Fixation
(BNF) play a major role in sustainable agriculture. In nature, BNF is restricted to certain
prokaryotic diazotrophs like bacteria, actinomycetes (Frankia) and blue -green algae

(Cyanobacteria) which are either free living or associated with plants. This biological
source accounts for 40-45 % of the total nitrogen fixed annually in the biosphere. By
using the available skill and back ground information, it is now possible to manipulate
the BNF system (bacteria and BGA) for biotechnological purposes. Several mutants for
antibiotic resistance, nitrogenase absence (nif-) oxygen sensitivities, heterotrophy and
altered heterocysts are now available and many strains can be grown clonally with ease.
Gene transfer is also possible now. Another recent approach is to extend the legume Rhizobium

symbiosis

to

Industrially,microorganisms

non
are

legumes

exploited

for

by
the

biotechnological
production

of

approach.
antibiotics,

pharmaceuticals, single cell protein, alcohol and beverages, organic acids, enzymes, dyes
etc. by proper biotechnological approaches; it is now possible to develop super strain for
augmenting the production. To be precise, microbes are sources of novel genes for any
biotechnological manipulation. Microbes have been used in food industry for centuries to
extend shelf life and confer traits that alter the flavor, texture or nutritional value of the
starting food materials. Improving the capability of microbes to perform their function
has changed from natural selection of strains to targeted improvement through
mutagenesis and the application of molecular biology techniques such as regulation and
deregulation of biochemical pathways in microbes, mutagenesis, etc:

Table 1. Some bacteria of biotechnological importance among 19 bacterial groups

Sl.
No

Group

Family

Genus

Process

1.

Gram
negative
aerobic

Pseudomonadaceae

Pseudomonas

rods/
cocci

Single Cell Protein

(SCP) From Methanol,
Oxidation of Steroids/
Hydrocarbon,
Polysaccharides
(Alginate); Oxidation Of
Alcohols
SCP From Methanol
Oxidation Of Methane

Methylomonadaceae

Methylomonas

Nonsymbiotic binding of
nitrogen

Methylococcus
Azotobacter
Azotobacteriaceae
2.

Gram
Enterobacteriaceae
negative
facultative
anaerobic

Escherichia

Many different
processes, production of
amino acid (lysine)

Aerobacter

Nucleotides, 2ketoglutaric acid,

pullulanase, 6aminopenicillanic acid,
recombinant rennet
Leaching of copper,
zinc, iron,manganese,
other sulphides

rods

3.

4.

Gram

negative
chemolithotrophic
bacteria
Methane
Methanobacteriaceae
producing
Nocardiceae
bacteria

Thiobacillus

Methano
bacterium
Nocardia

Methane from sewage

Oxidation of
Hydrocarbon, Steroids.

5.

Grampositive
cocci

Micrococcaceae

Micrococcus

Oxidation
Hydrocarbon,
starter culture

Streptococcaceae

Streptococcus

Production of lactic acid,

diacetyl; cheese and
fermented dairy product
starter

(lactococcus)

of
meat

Meat starter
Pediococcaceae

6.

Endospore Bacillaceae
forming
rods / cocci

pediococcus
leuconostoc

Bacillus

Clostridium
7.

8.

GramLactobacillaceae
Positive
asporogene
ous
rodshaped
bacteria
ActinoCorynebacterium
mycete
group
related
organisms
Propionibactericeae
Mycobacteriaceae

Lactobacillus

Bifidobacterium
Corynebacterium

Dextran
production;
cheese starter, wine
starter
(heterofermentation)
Antibiotics,
many
enzymes, amino acids,
and vitamins (B2,B12)
Butanol, acetone, butyric
acid, botulins
lactic acid, fermented
milk products, fermented
sausage and vegetables;
silage, spoilage of foods
bifidoyogurt,
bifidotablets
Oxidation of
Hydrocarbon, amino
acids,

Arthrobacter

transformation of
steroids

Cellulomonas

cellulose fermentation

Propionibacteria

vitamin B12 , propionic

acid, cheese
fermentation

Mycobacterium

Oxidation of
Hydrocarbon and
steroids

Cyanobacteria

Biological nitrogen
fixation
Beta- keratene

Dunalliella
Spirulina

Single Cell Protein

(SCP)

Chlorella
Plant biotechnology
For centuries, farmers have made improvements to crop plants through selective
breeding and hybridization- the controlled pollination of plants. Plant biotechnology is an
extension of this traditional plant breeding with one very important difference. Plant
biotechnology allows for the transfer of a great variety of genetic resources in a more,
precise, controlled manner.
Unlike traditional plant breeding, which involves the crossing of hundreds of
genes, plant biotechnology allows for the transfer of only one or a few desirable genes.
Hence it is possible to develop crops with specific beneficial traits and without
undesirable traits, such as those that would reduce crop yields.
Many of these beneficial traits in new plant varieties include

Resistance to pest and diseases and abiotic factors

Quality improvement tastier fruits-tomatoes with higher solids content

Nutrition enhancement- oil seeds that produce oils with low saturated fat content

Environmental protection.

Insect protection
Generally, insecticides are sprayed indiscriminately to control insects. This is not only
costly but also affects the environment in a larger extent. This could be managed by
biotechnological methods.

Bacillus thruingiensis (B.t.), a naturally occurring bacterium present in soil, is known for
its ability to control insect pests. Different strains of Bacillus thrugiensis control different
pests. First discovered in 1902, farmers have been using for decades as a biological
insecticide spray. B.t. produces a protein (cry protein) that disrupt the digestive system of
target insects, while remaining harmless to other insects, human, birds and other animals.
Now, through biotechnology, researchers are able to introduce B.t. gene into plants
thereby making them resistance to certain insect pests. By using this technology,
Monsonto has developed.
The following B.t. transgenics.
1. New leaf potato- resistance to Colorado beetle.
2. Bollgard

TM

cotton - resistance to bollworm, tobacco budworm and pink bollworm.

3. Yield gard TM corn - resistance to European corn borer.

4. Round up TM soyabean herbicide resistant.
Advantages

Ecofriendly- no air, groun water pollution

No disturbance to beneficial insects

More compatible to IPM

Offers season- long, effective control

Easy to use and cost effective.

Disease protection
Plant disease, including fungal and viral diseases, can devastate the yield and
quality of crop production. Using biotechnology tool such as introducing a small part of
DNA from a virus into the genetic make up of a plant, it is now possible to protect alfalfa,
canola, corn, cucumbers, grapes, papaya, potatoes, soybeans,squash and tomatoes from
viral disease. Similarly, by transferring chitinase gene plants are protected from fungal
disease.

To minimize the economic loss resulting from plant disease, farmers often must plant
more acreage than they expect to harvest. This extra acreage increases farmers planting,
fuel, water and fertilizer expenses, which must be passed on to the consumer. By using
transgenic crops, farmers can reduce resources used, such as the expense of labour, fuel,
pesticides, seed and equipment, to plant extra acres. This decreased planting will result
in increased soil conservation.
By introducing a gene or genes into a crop plant, many other advantageous features may
be possible. This includes

A genetic trait that controls the ripening of tomatoes, peppers and tropical fruits. This

trait allows time to ship crops long distances and results in tastier foods far from crops
native regions.

Potatoes and tomatoes developed with higher solids content. This trait offers

decreased processing costs because less energy is needed to extract water when
producing potato and tomato products. The higher solids content of potatoes holds the
potential to bring consumers lower fat French fries. Because oil replaces water during
frying. Process, potatoes with higher solids content (and less water) absorbs less oil.

Corn and soybeans with increased essential amino acid content- the building blocks

of protein. This trait can improve the quality of protein in food products and animal feed
made from these crops.

Naturally decaffeinated coffee

Corn and pees that retain their natural sweetness

Crops with modified fatty acid content allowing for the production of more healthful

oils

Biodegradable plastics developed from soybean polymers.

Improving the vit. A content in rice (Golden rice)

Fortifying rice with Fe and Zn.

Many existing food products are currently being modified through biotechnology as
follows.
Sl.

Product

Genetic

No.
1.

Modification
Tomatoes, peas, peppers, tropical Controlled

Allow shipping of vine

fruits, broccoli, rasp berries,

ripened tomatoes, improve

ripening

melons
2.

3.
4.

Purpose

quality, shelf life, food

processing.
Reduce insecticide use

Tomatoes, potatoes, corn,

Insect

lettuce, coffee, cabbage family,

resistance

apples
Peppers, tomatoes, cucumbers

Fungal

Reduce fungicide use

Potatoes, tomatoes, squash,

resistance
Viral

Reduce diseases caused by

cucumbers, corn, oilseed, rape

resistance

plant viruses and since insect

(canola), soybeans, grapes.

carry viruses, reduce

5.

Soybeans, tomatoes, corn,

Herbicide

insecticide use
Improve weed control

6.

canola, wheat
Corn, sunflowers, soybeans, rice

tolerance
Improved

Increase the amount of

etc.

nutrition

essential amino acids,

vitamins or other nutrients in

7.

8.

Canola, palm oil,

Canola, peanuts

Improved

the host plant

Increase the amount of

nutrition

unsaturated oils in the plant by

Heat stability

affering the oil.

Improve processing quality
permit new food uses for

Coffee

Low caffeine

healthier oils
Naturally decaffeinate coffee

Corn, peas

content
Controlled

Retain sweetness during entire

11.

Rice

starch
Vit. A

shelf life
Reduce incidence of blindness

12.

Rice pulses, corn

More protein

in developing countries.
Complement essential amino

9.
10.

acids; tolerate drought and

13.

Cassava

Reduce cyano

acidic soils
Food safety non toxic

geneic
14.

Potato

glucosides
Modification

Withstand frost

of acid
15.

Corn and soybeans

production
More protein

Increased levels of lysine and

methionine for improved

Tomatoes

Higher

animal feed.
Antioxidant

17.

Strawberries

lycopene
Higher ellagic

Anti cancer agent.

18.

Garlic

acid
More allicin

Lower cholesterol

16.

One of the most striking differences between traditional breeding and biotechnological
approach is that the source of genetic material need not come from the same species.
Hence, it is possible to exchange genetic information between bacteria, plants and
animals (including humans). New technologies even allow scientists to decide in which
part of the plant tissue, a trait should be expressed (Tissue specific expression) such as
the pulp versus the skin of the apple.

Advantages of biotechnology over irraditional breeding

1.

Transfer of a single specific gene of insect from any plant and move it

to genetic code of another plant instead of transferring thousands of genes in traditional

breeding.
2.

Reduction of breeding time substantially (from 10-12 years to 4-5

years).
3.

Replacing high dose animal test to low dose test for toxicity

studies.
4.

Possibility of transferring desired gene from any source.

Noval produce such as seedless tomatoes, squash, egg plant, peppers, strawberries,
melons and cherries would be attractive to many consumers. Additionally these fruits
would have more taste due to increased total soluble sugar than seeded fruits and may be
of economic value to the processing industry as well. Attempts are now to produce sweet
protein from the African vine Pentadiplandra brazzeana. This heat stable protein is 500
times as sweet as sucrose at higher concentration and as much as 2000 times as sweet in a
2 % (by weight) solution. Lacking bitterness, it has a lot of potential as an alternative low
energy sweetener.
Animal biotechnology
In animal biotechnology programme, transgenic technologies are used to produce humanvaluable proteins in genetically modified animals. Human serum albumin, a major
protein constituent of the blood (which is used to treat emergency blood losses and
chronic blood deficiency), and factor VIII can essential constituent of the body response
to repair injuries of blood vessels), can be produced in goat milk to replace the blood
derived products that are currently threatened by risks of AIDS and other diseases.
Human growth hormone is produced in goat milk by serumtech Ltd.(Israel) and
pharmaceutical proteins ltd (UK) uses this technology to produce anti- -transferin
antibody in sheep. Gene pharming Ltd (The Netherlands) uses cows for the production of

lactoferin, collagen, and crythropoitin and the Red Cross is using swine to develop
protein c for blood coagulation.
The first FDA approved application of biotechnology to animal production was the
use of recombinant somatotrophin (rbst) in dairy cows. bst, a protein hormone found
naturally in cows, is necessary for milk production. This can be isolated from cows,
inserted into bacterial cells, and produced in large quantities by those cells. The bst can
then be recovered, purified and injected into dairy cows. The injection has been shown to
increase the milk production by 10-25 %FDA gave approval in 1993 based on the
following aspects.
1. bst is species- specific for cows
2. bst is a protein that is digested in the intestinal tract of human beings and cows.
3. Milk contains naturally and supplementation does not increase the amount of bst to
levels beyond normal ranges
4. bst supplementation does not change milk composition and
5. bst has not been found to cause growth promoting activity in a variety of species.
Porcine somatotropin (pst), a hormone similar to bst but active in hogs, is a repartioning
agent. It helps to redirect all dietry energy for lean muscle tissue production, resulting
increased pork carcass learnness and reduces fattiness. Although pst has not yet been
approved in USA, it has been approved in European countries.
Other uses of biotechnology in animal production
Development of vaccines to protect animal from disease (eg.sine pseudorabies)
Production of several calves from one embryo (cloning)
Artificial insemination
Improvement of growth rate and /or feed efficiency.(GM. Salmon that grows more
rapidly to the marketed size than regular Salmon)
Production of transgenic animals or animals with altered genetic make-ups

Developments of biotechnology application in animal sciences may be slow due to

greater difficulties in animal genetic engineering as well as serial and ethical concerns of
consumers. Most transgenic animal research is for medical applications, as in the case of
Dolly (cloned sheep), where scientists are investigating cystic fibrosis disease.