18:Biotechnology & Genetic Engineering

Unit 18: Biotechnology and genetic modification

18.1 Biotechnology
1 Explain the role of yeast in the production of bread and ethanol
2 Understand that bacteria are useful in biotechnology and genetic modification due
to their rapid reproduction rate and their ability to make complex molecules
3 Discuss why bacteria are useful in biotechnology and genetic modification, limited
to:
(a) no ethical concerns over their manipulation and growth
(b) presence of plasmids
4 Describe how fermenters can be used for the large-scale production of useful
products by bacteria and fungi, including the conditions that need to be
controlled, limited to: temperature, pH, oxygen, nutrient supply and waste
products
5 Describe the use of:
(a) enzymes in biological washing powders
(b) pectinase for fruit juice production
(c) lactase for lactose-free milk
18.2 Genetic modification
1 Describe genetic modification as changing the genetic material of an organism by
removing, changing or inserting individual genes
2 Understand that the gene that controls the production of human insulin has been
inserted into bacterial DNA, for commercial production of insulin
3 Outline the use of genetic modification in crop plants by inserting genes:
(a) to confer resistance to herbicides
(b) to confer resistance to insect pests
(c) to provide additional vitamins
4 Discuss potential advantages and risks of genetic modification, limited to
modifying crop plants and bacteria.

Learn
 How and why is yeast important in biotechnology?
 How are bacteria useful in biotechnology and genetic modification?
 Why are bacteria useful in biotechnology and genetic modification?
 How are fermenters used for large-scale production of biotechnology
products?
 What conditions need to be controlled in a fermenter?
 How are enzymes used in biotechnology?
 What is the role of anaerobic respiration in yeast during the production of
biofuels and in bread-making?
 What is the role of pectinase in fruit juice production?
 What is the role of biological washing powders that contain enzymes?
 What is the role of lactase in lactose-free milk production?

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Genetic modification the process in which the genes from one organism
are transferred to another organism of different species.
Biotechnology: the application of biological organisms, systems or
processes to manufacturing and service industries.

Use of Yeast (fungi) in Biotechnology:

 Yeast can produce ethanol and antibiotics during anaerobic respiration.
 They are unicellular and have a faster rate of reproduction.
 No ethical concern

Fermentation:
Fermentation is a number of enzyme controlled reactions occurring inside
the microorganisms such as yeast to release energy.
e.g. the anaerobic respiration of glucose to ethanol.
Fermentation of microorganisms is required for commercial production of-
1. Biofuels 5. Antibiotic
2. Bread 6. Commercial enzymes
3. Alcoholic beverage
4. Single cell protein

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Use of fungi in Biotechnology:

1. Bread production: baking
o Flour, water, salt, oil and yeast are mixed to make a dough.
o Yeast has no enzymes for digesting the starch in flour, but adding
water activates the amylases present in flour. The amylase digests
some of the starch to sugar.
o The yeast then ferments the sugar to ethanol and carbon dioxide.
o The dough is repeatedly folded and stretched (kneaded) and then left
for few hours at a temperature of about 27 °C.
o The carbon dioxide makes bubbles in the bread dough. These bubbles
make the bread light in texture
o A protein called gluten gives the dough a sticky, elastic texture, which
holds the bubbles of gas.
o The carbon dioxide bubbles build up, making the dough rise to about
double its volume.
o The dough is kneaded again and baked at about 200 °C.
o During baking, high temperatures kill the active yeast and the ethanol
evaporates.

2. Alcohol production: brewing

 Yeast is mixed with sugar sources from fruit or grain extracts
 The mixture is then taken in a fermenter and kept in a constant
temperature and pH.
 In absence of O2 yeast respire on sugar anaerobically, producing
alcohol and CO2
 The alcohol is then collected from the fermenter through distillation.

Ethanol as Biofuels
 Ethanol acts as renewable source of energy which can be used in
combustion as a biofuel.
 It produces less pollution than petrol.
 Ethanol can be produced from fermented sugar or spare grain by yeast.
 Since 2013, 94% of new cars in Brazil use a mixture of petrol and
ethanol.
 Biofuels are not yet economical to produce as,
o They are expensive and require a huge amount of energy during
the growth and processing of sugar.
o May harm the environment. e.g. soy beans or oil palms destroy
forest , removing the habitats and endangere species.

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Importance of Bacteria in Biotechnology:

● Can be used by humans to produce foods and other useful substances.
● Yoghurt production
● Cheese production
● Commercial enzyme production
● Insulin production
● Sewage disposal depends on bacteria in the filter beds to form the
basis of the food chain that purifies the effluent.

Why bacteria are useful in biotechnology and genetic modification:

 capable of producing complex molecules
 reproduce rapidly
 they can be grown and manipulated without raising ethical concerns
 presence of plasmids for complex molecule formation
 They have a genetic code/plasmid that is the same as all other
organisms, so scientists can transfer genes from other animals or
plants into bacterial DNA.

Fermenter: A bioreactor

1. Cooling system:
● Cold water is pumped through a cooling Jacket surrounding the
fermenter.
● It removes the excess heat produced during fermentation.
● A sensor is used to measure the temperature inside the fermenter.

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2. Aeration system: O2 is needed for aerobic respiration in

microorganisms.
Proper mixing and adequate aeration help the microorganisms to multiply
rapidly.
 Sparger: a metal ring with tiny holes through which sterile air is
passed inside the fermenter under high pressure.
 Impeller:
○ a stirring device that mixes air bubbles well with the nutrient
broth.
○ It ensures even distribution of oxygen and nutrients.
○ Continuous stirring prevents agglutination of microorganisms.
3. pH controller:
a. Maintains an optimum pH for maximum growth of the
microorganisms.
b. A sensor also detects the pH of the broth and an acid/base
reservoir (buffer) to adjust the pH.
4. Nutrients: the broth contain-
a. Carbon and energy source, e.g., glucose
b. Nitrogen source, e.g., amino acid
c. Essential mineral salts

Use of enzymes in biotechnology:

The role of lactase in yoghurt production
 Lactase is used in the commercial production of yoghurt to speed up the
process and enhance its taste.
 The starter cultures in the production of yoghurt are Lactobacillus
bulgaricus and Streptococcus thermophilus.
 The bacterial culture is added to pasteurised milk at 42 °C.
 The bacteria ferment lactose, the milk sugar into lactic acid.
 The acid gradually increases the acidity of the fermented milk to a pH of
4.5.
 The acidity causes the milk to clot, forming yoghurt and provide the
flavour.

Fruit Juice Production

 Pectinases are enzymes extracted from fungi and used to separate the
juices from fruit like apples.
 Pectinase is used to make fruit juice more transparent.
● Fruit juice is produced by squeezing the fruits.

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● Chopping the fruit up before squeezing helps to release a lot of juice.

But different polysaccharides are released, which make the juice
cloudy.
● Pectinase break down pectin, the jelly-like substance that sticks plant
cell walls to each other.
● Pectinases break these down to make the juice clearer. The sugars
produced also make the juice sweeter.

Biological Washing Powders

● Proteases and lipases are manufactured for the use in the food and
textile industries.
● Biological washing powders contain different types of digesting
enzymes to remove the organic food stains.
● Advantages of using biological washing powders are:
○ Quickly breaking down large, insoluble molecules into smaller,
soluble ones that will dissolve in washing water.
○ They are effective at lower temperatures, meaning less
energy and cost is required
○ They can be used to clean delicate fabrics that would not be
suitable for washing at high temperatures
● If biological washing powder denature at high temperature

Lactose-Free Milk
● Lactose is a milk sugar.
● Human babies are born with the ability to produce lactase, the enzyme
that breaks down lactose
● The people who lose the ability to produce lactase become lactose
intolerant and react badly to the lactose in milk as well as milk products.
● Symptoms of lactose intolerance include nausea, flatulence and
diarrhea.
● Milk can be made lactose free by treating milk with the enzyme
lactase.

Antibiotics
One of the richest sources of antibiotics is Actinomycetes bacteria that look
like microscopic mould fungi.
 The actinomycete Streptomyces produces the antibiotic streptomycin.
 Penicillin is produced by the mutant forms of a different species of
Penicillium from the fungus.

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The antibiotics are chemically altered in the laboratory to make them more
effective and suitable for the use with different diseases. E.g. Ampicillin,
methicillin and oxacillin.
Antibiotics attack bacteria by inhibiting the production of the cell wall to stop
their reproduction or burst the cell open. Some antibiotics affect protein
synthesis, to stop the bacterial growth.
Antibiotics do not damage human cells, as they do not have cell walls and
their protein production machinary are different.

Genetic Engineering
Genetic engineering is changing the genetic material of an organism by
removing, changing or inserting individual genes from another organism.

● The organism receiving the genetic material is said to be ‘genetically

modified’, or a ‘transgenic organism’

● The DNA of the organism that now contains DNA from another
organism is known as ‘recombinant DNA’

● Examples of genetically modified organisms are:

○ The gene for human insulin has been inserted into bacteria
which then produce human insulin which can be collected and
purified for medical use for diabetics
○ Crop plants, such as wheat and maize, have been genetically
modified to contain a gene from a bacterium that produces a
poison that kills insects, making them resistant to insect
pests such as caterpillars
● Crop plants have also been genetically modified to make them
resistant to certain herbicides (chemicals that kill plants), meaning
that when the herbicide is sprayed on the crop it only kills weeds and
does not affect the crop plant.
● Genetically modified plants can be produced with additional
vitamins, eg ‘golden rice’ produce a chemical that is turned into
vitamin A in the human body, which could help prevent deficiency
diseases in certain areas of the world.

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Process of Genetic Engineering

● The desired gene that is to be inserted is located in the original
organism
● Restriction enzymes are used to isolate the required gene, leaving it
with ‘sticky ends’.
● A bacterial plasmid is cut by the same restriction enzyme leaving it
with corresponding sticky ends.
● Restriction enzymes cut DNA strands at specific sequences to form
‘sticky ends’
● The plasmid and the isolated gene are joined together by DNA ligase
enzyme
● If two pieces of DNA have complementary sticky ends, DNA ligase
will join them to form a single molecule of DNA
● The genetically engineered plasmid is then inserted into a bacterial
cell
● When the bacteria reproduce the recombinant plasmids are copied
and quickly spread as the bacteria multiply
● The plasmids will then express all the gene and make the protein
● The genetically engineered bacteria can be placed in a fermenter to
reproduce quickly in controlled conditions and make large quantities
of the desired protein

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● Bacterial plasmids are extremely useful for genetic engineering purposes

because:
○ They contain the same genetic code as the other organisms,
meaning they can produce the same proteins
○ There are no ethical concerns
○ The plasmids in bacteria are easy to remove and manipulate to
insert genes into them and then place back inside the bacterial cells.

The basis of genetic engineering is the production of recombinant DNA. For

example, if a bacterium receives the human gene that codes for insulin
production, it will make human insulin.

Further research found the following enzymes that were able to do this:
◾ Restriction enzymes that cut DNA molecules at specific points. Different
restriction enzymes cut DNA at different places. They can be used to cut out
specific genes from a molecule of DNA.
◾ Ligases (or DNA ligases) are enzymes that join the cut ends of DNA
molecules.

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Advantages of GM Crops:
(i) More tolerance to biotic and (iv) Enhanced nutritional value
abiotic stresses (hormones, of food, e.g., vitamin-A enriched
nutrition, heat etc) rice (Golden rice), protein
(ii) Resistance to pests. enriched vegetables like
(iii) Increases fertility of soil spinach, brocolli etc.

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Disadvantage of GM crops are:

(a) Transgenic genes in GM crops endanger native species.
(b) They cause damage to natural environment by reducing
biodiversity.
(c) They may cause human health problems by altered genes.

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