Biotechnology

and
Industry
Applications

Improvement in Fermentation Products.This

achievement can be done in different ways – by
selection of improved strain, by transgene application
into the microorganism, by using cheaper raw material,
by manipulation of medium constituent as well as by
simulation of the reactor (adjustment of different
cultural conditions like pH, temp., etc.).Products of
microbial fermentation include primary metabolites,
secondary metabolites, enzymes, proteins, capsular
polysaccharides and cellular biomass (single cell
protein).

Microbial Production of Synthetic Fuels.Important

fuels can be produced by using many microbes which
include ethanol, methane, hydrogen and hydrocarbons.
Zymomonas mobilis produces ethanol twice as rapidly
as yeasts from carbohydrates. Methane which is used in
various industrial purposes can be produced by
Clostridia, Bacteriodes, Sclenomonas, Butyrovibrio, etc.
from the waste.

Microbial Biomass and Single Cell Protein Production.

Microbial product of commercial significance is the
microbial biomass (the microbial cells themselves), e.g.,
commercially produced yeast cells, bacteria
(Methylophilus methylotrophus), flavoring cheese from
fungal biomass (Penicillium roquefortii).
 Microbial Mining or Bioleaching:The process of
bioleaching recovers metals from ores which are not
suitable for direct smelting because of their low content.
The application of bioleaching process is of particular
interest in case of uranium ore. Thiobacillus ferroxidans
is the commonest organism which is involved in case of
copper and uranium ore processing.

Production of Secondary Metabolites from Cultured

Plant Cell.In recent years it has been shown that
spectrum of compounds can be produced in culture
which is beyond the ability of whole plants. By using
different precursors several novel compounds of
biomedical importance can be obtained.Pharmaceutical
compounds like shikonin is being produced as
secondary products with the use of two-stage bioreactor
by stimulating the growth phase with the application of
different growth regulators.

Serpentine can be obtained from Catharanthus,

pseudoephedrine from Ephedra. Plant cells also can be
used to accomplish certain changes in the structure and
composition of some industrially important chemicals.
This conversion by means of a biological system is
termed as biotransformation, e.g., digoxin, a
cardiovascular drug, produced from digitoxin obtained from Digitalis lanata.
 Applications

Improved Yield from Crops. Using biotechnology techniques,

one or two genes may be transferred into a crop to give a new
trait to that crop. This is done in the hope of increasing its yield.
However, these increases in yield have proved to be difficult to
achieve. Current genetic engineering techniques work best for
single gene effects - that is traits inherited in a simple
Mendelian fashion. Many of the genetic characteristics
associated with crop yield, such as enhanced growth, are
controlled by a large number of genes, each of which just has a
slight effect on the overall yield. There is, therefore, still much
research, including plant genetic research, to be done in this
area.

Reduced Vulnerability to Environmental Stresses.Crops are

obviously dependent on environmental conditions. Drought can
destroy crop yields, as can too much rain or floods. But what if
crops could be developed to withstand these harsh conditions?
Biotechnology will allow the development of crops containing
genes that will enable them to
withstand biotic and abiotic stresses. For example, drought and
excessively salty soil are two significant factors affecting crop
productivity. But there are crops that can withstand these harsh
conditions. Why? Probably because of that plant's genetics. So
biotechnologists and plant geneticists are studying plants that
can cope with these extreme conditions, trying to identify and
isolate the genes that control these beneficial traits. The genes
could then be transferred into more desirable crops, with the
hope of producing the same phenotypes in those crops.
 Increased Nutritional Qualities of Crops. Maybe you’ve heard
over and over that eating beans is good for you. True? Well,
maybe. But what if it were genetically possible to increase the
nutritional qualities of food? One would think that would be
beneficial to society. So, can biotechnology be used to do just
that? Scientists are working on modifying proteins in foods to
increase their nutritional qualities. Also, proteins in legumes
and cereals may be transformed to provide all the amino acids
needed by human beings for a balanced diet.
Improved Taste, Texture or
Appearance of Food.Have you
ever gone to the grocery store,
bought some fruit and never gotten around to eating it? Maybe
you haven’t, but maybe your parents have. Modern
biotechnology can be used to slow down the process of spoilage
so that fruit can ripen longer on the plant and then be
transported to the consumer with a still reasonable shelf life.
This is extremely important in parts of the world where time
from harvest to the consumer may be longer than in other areas.
In addition to improving the taste, texture and appearance of
fruit, it will also extend the usable life of the fruit. As the
world population grows and grows, this may become a fairly
important issue. Extending the life of fruit can expand the market
for farmers in developing countries due to the reduction in spoilage. This has successfully been
demonstrated with the tomato. The first genetically modified food product was a tomato which
was transformed to delay its ripening. Researchers in Indonesia, Malaysia, Thailand, Philippines
and Vietnam are currently working on developing other delayed ripening fruits, such as the
papaya.

Reduced Dependence on Fertilizers, Pesticides and Other

Agrochemicals.There is growing concern regarding the use
of pesticides in agriculture. Therefore, many of the current
commercial applications of modern biotechnology in
agriculture are focused on reducing the dependence of
farmers on these chemicals. For example, Bacillus
thuringiensis (Bt) is a soil bacterium that produces a protein
that can act as an insecticide, known as the Bt toxin. But it is
a naturally occurring protein, not a foreign chemical. Could
this protein be used in crops instead of pesticides?
Traditionally, an insecticidal spray has been produced from
these bacteria. As a spray, the Bt toxin is in an inactive state
and requires digestion by an insect to become active and
have any effect. Crop plants have now been engineered to contain and express the genes for the
Bt toxin, which they produce in its active form. When an insect ingests the transgenic crop, it
stops feeding and soon thereafter dies as a result of the Bt toxin binding to its gut wall. Bt corn is
now commercially available in a number of countries to control corn borer (an insect like a moth
or butterfly), which is otherwise controlled by insecticidal spraying.
 History

About 10,000 years BC, people harvested their food from the natural biological diversity
that surrounded them, and eventually domesticated crops and animals. During the process of
domestication, people began to select better plant materials for propagation and animals for
breeding, initially unwittingly, but ultimately with the intention of developing improved food
crops and livestock. Over thousands of years farmers selected for desirable traits in crops, and
thus improved the plants for agricultural purposes. Desirable traits included crop varieties (also
known as cultivars, from "cultivated varieties") with shortened growing seasons, increased
resistance to diseases and pests, larger seeds and fruits, nutritional content, shelf life, and better
adaptation to diverse ecological conditions under which crops were grown.
Over the centuries, agricultural technology developed a broad spectrum of options for food, feed,
and fiber production. In many ways, technology reduces the amount of time we dedicate to basic
activities like food production, and makes our lives easier and more enjoyable. Everyone is
familiar with how transportation has changed over time to be more efficient and safer (Figure 1).
Agriculture has also undergone tremendous changes, many of which have made food and fiber
production more efficient and safer (Figure 1). For example in 1870, the total population of the
USA was 38,558,371 and 53% of this population was involved in farming; in 2000, the total
population was 275,000,000 and only 1.8% of the population was involved in farming. There are
negative aspects to having so few members of society involved in agriculture, but this serves to
illustrate how technological developments have reduced the need for basic farm labor.

Agriculturalists started conducting selective breeding of crops before having a thorough

understanding of the basis of genetics. Gregor Mendel's discoveries explaining how traits pass
from parents to offspring shed new light on the matter. Mendel's work showed
that genes separate during the formation of gametes, and unite randomly during fertilization; he
also showed that genes are transmitted independently of one another to offspring. This
understanding of the way that plants and animals acquire traits form parents created the potential
for people to selectively breed crops and livestock. Gregor Mendel's discovery revolutionized
agriculture by launching the development of selective cross breeding with a comprehensive
understanding of the underlying mechanisms of inheritance.
Selective Cross Breeding
In traditional plant breeding, new varieties are developed either by selecting plants with desirable
characteristics or by combining qualities from two closely related plants through selective
breeding. These features may for example be resistance to a particular pest or disease, or
tolerance to climatic conditions. Pollen with the genes for a desired trait is transferred from
plants of one crop variety to the flowers of another variety with other desirable traits. Eventually,
through careful selection of offspring, the desired trait will appear in a new variety of plants.
Traditional plant breeding has produced numerous highly successful new varieties of crops over
the centuries. There have also been many less than successful crosses made. In traditional
breeding, crosses are often made in a relatively uncontrolled manner. The breeder chooses the
parents to cross, but at the genetic level, the results are unpredictable. DNA from the parents
recombines randomly, and desirable traits such as pest resistance may be bundled with
undesirable traits, such as lower yield or poor quality. The parent plants must be closely related
to produce offspring. Traditional breeding programs are time-consuming, often taking decades to
produce new viable crop varieties, and labor-intensive. A great deal of effort is required to
separate undesirable from desirable traits, and this is not always economically practical. Many
potential benefits are lost along the way, as plants that fail to demonstrate the introduced
characteristics are discarded. Traditional plant breeding takes on average 12-15 years to produce
a new crop variety.
Classical Breeding with Induced Mutation
Mutations (Figure 2) are changes in the genetic makeup of a plant. Mutations occur naturally and
sometimes result in the development of new beneficial traits. In 1940, plant breeders learned that
they could make mutations happen faster with a process called mutagenesis. Radiation or
chemicals are used to change the plant's DNA, the basic molecular system of all organisms'
genetic material. The goal is to cause changes in the sequence of the base pairs of DNA, which
provide biochemical instructions for the development of plants. Resultant plants may possess
new and desirable characteristics through this modification of their genetic material. During this
process, plant breeders must grow and evaluate each plant from each seed produced. More than
2,500 plant varieties (including rice, wheat, grapefruit, lettuce and many fruits) have been
developed using radiation mutagenesis. Induced mutation breeding was widely used in the
United States during the 1970's, but today few varieties are produced using this technique. As
our understanding of genetics developed, so new technologies for plant variety development
arose. Examples of these that are used today include genetic marker assisted breeding, where
molecular markers associated with specific traits could be used to direct breeding programs, and
genetic engineering. Some of the significant steps leading to the current state of the art are
explained below.

1. Discovery by Watson and Crick: structure of DNA, 1953: Another milestone in the

development of understanding of genetics and how genes function, was the discovery of the
structure of DNA (the basis of genes), and how DNA works. Two scientists, James Watson and
Francis Crick made this discovery (Pray 2008), considered to be one of the most significant
scientific works in biology, largely through synthesis of the work of other scientists. Their work
contributed significantly to understanding what genes were.

2. Discovering genes that move (transposons): Transposons are sections of DNA-genes-that

move from one location to another on a chromosome. Transposons have been referred to as
"jumping genes", genes that are able to move around. Interestingly, transposons may be
manipulated to alter the DNA inside living organisms. Barbara McLintock (1950) discovered an
interesting effect of transposons. She was able to show how the changes in DNA caused by
transposons affected the color of maize kernels.

3. Tissue culture and plant regeneration: Another significant development in technology that

was important for plant breeding was the development of micropropagation techniques, known
as tissue culture (Thorpe 2007). Tissue culture permits researchers to clone plant material by
excising small amounts of tissue from plants of interest, and then inducing growth of the tissue
on media, to ultimately form a new plant. This new plant carries the entire genetic information of
the donor plant. Exact copies of a desired plant could thus be produced without depending on
pollinators, the need for seeds, and this could all be done quickly.
4. Embryo rescue: Often when distantly related plant species are hybridized are crossed, the
embryos formed following fertilization will be aborted. The development of embryo rescue
technology permitted crop breeders to make crosses among distantly related varieties, and then to
save the resulting embryos and then grow them into whole plants through tissue culture.

5. Protoplast fusion: Protoplasts are cells that have lost their cell walls. The cell wall can be
removed either by mechanical means, or by the action of enzymes. They are left with only a cell
membrane surrounding the cell. Protoplasts can be manipulated in many ways that can be used in
plant breeding. This includes producing hybrid cells (by means of cell fusion) and using
protoplasts to introduce new genes into plant cells, which can then be grown using tissue culture
techniques (Thorpe 2007).

6. Genetic engineering: Building on the above discoveries into the 1980s, advances in the field
of molecular biology provided scientists with the potential to purposefully transfer DNA between
organisms, whether closely or distantly related. This set the stage for potentially extremely
beneficial advancement in crop breeding, but has also been very controversial.

Genetic Engineering of Organisms

The basic structure of DNA is identical in all living things. In all organisms, different
characteristics are determined by the sequence of the DNA base pairs. Biotechnology has
developed to the point where researchers can take one or more specific genes from nearly any
organism, including plants, animals, bacteria, or viruses, and introduce those genes into
the genome of another organism. This is called recombinant DNA technology (Watson et al.
1992). In 1978, the first commercial product arising from the use of recombinant DNA
technology gene transfer was synthetic insulin. Pig and cattle pancreatic glands were previously
the only way of producing insulin for human use. In 1988, chymosin (known as Rennin) was the
first enzyme produced from a genetically modified source-yeast-to be approved for use in food.
Previously this enzyme for cheese production was obtained from cows' stomach linings.
In agricultural biotechnology, changes are made directly to the plant's genome. Once the gene
that determines a desirable trait is identified, it can be selected, extracted, and transferred directly
into another plant genome (Figure 3). Plants that have genes from other organisms are referred to
as transgenic. The presence of the desired gene, controlling the trait, can be tested for at any
stage of growth, such as in small seedlings in a greenhouse tray. A breeder can thus quickly
evaluate the plants that are produced and then select those that best express the desired trait.
Producing new varieties of crops through genetic engineering takes about 10 years on average.
 History

With ancestral roots in industrial microbiology that date back centuries, the new
biotechnology industry grew rapidly beginning in the mid-1970s. Each new scientific advance
became a media event designed to capture investment confidence and public support.
[15]
 Although market expectations and social benefits of new products were frequently overstated,
many people were prepared to see genetic engineering as the next great advance in technological
progress. By the 1980s, biotechnology characterized a nascent real industry, providing titles for
emerging trade organizations such as the Biotechnology Industry Organization (BIO).
The main focus of attention after insulin were the potential profit makers in the pharmaceutical
industry: human growth hormone and what promised to be a miraculous cure for viral
diseases, interferon. Cancer was a central target in the 1970s because increasingly the disease
was linked to viruses.[14] By 1980, a new company, Biogen, had produced interferon through
recombinant DNA. The emergence of interferon and the possibility of curing cancer raised
money in the community for research and increased the enthusiasm of an otherwise uncertain
and tentative society. Moreover, to the 1970s plight of cancer was added AIDS in the 1980s,
offering an enormous potential market for a successful therapy, and more immediately, a market
for diagnostic tests based on monoclonal antibodies. [22] By 1988, only five proteins from
genetically engineered cells had been approved as drugs by the United States Food and Drug
Administration (FDA): synthetic insulin, human growth hormone, hepatitis B vaccine, alpha-
interferon, and tissue plasminogen activator (TPa), for lysis of blood clots. By the end of the
1990s, however, 125 more genetically engineered drugs would be approved.[22]
The 2007–2008 global financial crisis led to several changes in the way the biotechnology
industry was financed and organized. First, it led to a decline in overall financial investment in
the sector, globally; and second, in some countries like the UK it led to a shift from business
strategies focused on going for an initial public offering (IPO) to seeking a trade saleinstead.
[23]
 By 2011, financial investment in the biotechnology industry started to improve again and by
2014 the global market capitalization reached $1 trillion.[23]
Genetic engineering also reached the agricultural front as well. There was tremendous progress
since the market introduction of the genetically engineered Flavr Savr tomato in 1994.[22] Ernst
and Young reported that in 1998, 30% of the U.S. soybean crop was expected to be from
genetically engineered seeds. In 1998, about 30% of the US cotton and corn crops were also
expected to be products of genetic engineering.[22]
Genetic engineering in biotechnology stimulated hopes for both therapeutic proteins, drugs and
biological organisms themselves, such as seeds, pesticides, engineered yeasts, and modified
human cells for treating genetic diseases. From the perspective of its commercial promoters,
scientific breakthroughs, industrial commitment, and official support were finally coming
together, and biotechnology became a normal part of business. No longer were the proponents
for the economic and technological significance of biotechnology the iconoclasts. [1] Their
message had finally become accepted and incorporated into the policies of governments and
industry.
 Date Events

 7000 BCE  Chinese discover fermentation through beer making.

 6000 BCE – Yogurt and cheese made with lactic acid-producing bacteria by various
people.
 4000 BCE – Egyptians bake leavened bread using yeast.
 500 BCE – Moldy soybean curds used as an antibiotic.
 250 BCE – The Greeks practice crop rotation for maximum soil fertility[
 100 CE – Chinese use chrysanthemum as a natural insecticide.

 1663 – First recorded description of living cells by Robert Hooke.

 1677 – Antonie van Leeuwenhoek discovers and describes bacteria and protozoa.
 1798 – Edward Jenner uses first viral vaccine to inoculate a child from smallpox.
 1802 – The first recorded use of the word biology.
 1824 – Henri Dutrochet discovers that tissues are composed of living cells.
 1838 – Protein discovered, named and recorded by Gerardus Johannes Mulder and Jöns
Jacob Berzelius.
 1862 – Louis Pasteur discovers the bacterial origin of fermentation.
 1863 – Gregor Mendel discovers the laws of inheritance.
 1864 – Antonin Prandtl invents first centrifuge to separate cream from milk.
 1869 – Friedrich Miescher identifies DNA in the sperm of a trout.
 1871 – Ernst Hoppe-Seyler discovers invertase, which is still used for making artificial
sweeteners.
 1877 – Robert Koch develops a technique for staining bacteria for identification.
 1878 – Walther Flemming discovers chromatin leading to the discovery of chromosomes.
 1881 – Louis Pasteur develops vaccines against bacteria that
cause cholera and anthrax in chickens.
 1885 – Louis Pasteur and Emile Roux develop the first rabies vaccine and use it
on Joseph Meister.

 1919 – Károly Ereky, a Hungarian agricultural engineer, first uses the word
biotechnology. 1928 – Alexander Fleming notices that a certain mould could stop the
duplication of bacteria, leading to the first antibiotic: penicillin.
 1933 – Hybrid corn is commercialized.
 1942 – Penicillin is mass-produced in microbes for the first time.
 1950 – The first synthetic antibiotic is created.
 1951 – Artificial insemination of livestock is accomplished using frozen semen.
 1952 – L.V. Radushkevich and V.M. Lukyanovich publish clear images of 50 nanometer
diameter tubes made of carbon, in the Soviet Journal of Physical Chemistry.
 1953 – James D. Watson and Francis Crick describe the structure of DNA.
 1958 – The term bionics is coined by Jack E. Steele.
 1964 – The first commercial myoelectric arm is developed by the Central Prosthetic
Research Institute of the USSR, and distributed by the Hangar Limb Factory of the UK.
 1972 – The DNA composition of chimpanzees and gorillas is discovered to be 99%
similar to that of humans.
 1973 – Stanley Norman Cohen and Herbert Boyer perform the first
successful recombinant DNA experiment, using bacterial genes.[4]
 1974 – Scientist invent the first biocement for industrial applications.
 1975 – Method for producing monoclonal antibodies developed by Köhler and César
Milstein.
 1978 – North Carolina scientists Clyde Hutchison and Marshall Edgell show it is possible
to introduce specific mutations at specific sites in a DNA molecule.[5]
 1980 – The U.S. patent for gene cloning is awarded to Cohen and Boyer.
 1982 – Humulin, Genentech's human insulin drug produced by genetically engineered
bacteria for the treatment of diabetes, is the first biotech drug to be approved by the Food and
Drug Administration.
 1983 – The Polymerase Chain Reaction (PCR) technique is conceived.
 1990 – First federally approved gene therapy treatment is performed successfully on a
young girl who suffered from an immune disorder.
 1994 – The United States Food and Drug Administration approves the first GM food: the
"Flavr Savr" tomato.
 1997 – British scientists, led by Ian Wilmut from the Roslin Institute, report
cloning Dolly the sheep using DNA from two adult sheep cells.
 1999 – Discovery of the gene responsible for developing cystic fibrosis.
 2000 – Completion of a "rough draft" of the human genome in the Human Genome
Project.

 2001 – Celera Genomics and the Human Genome Project create a draft of the human

genome sequence. It is published by Science and Nature Magazine.
 2002 – Rice becomes the first crop to have its genome decoded.
 2003 – The Human Genome Project is completed, providing information on the locations
and sequence of human genes on all 46 chromosomes.
 2008 – Japanese astronomers launch the first Medical Experiment Module called "Kibo",
to be used on the International Space Station.
 2009 – Cedars-Sinai Heart Institute uses modified SAN heart genes to create the
first viral pacemaker in guinea pigs, now known as iSANs.
 2012 – Thirty-one-year-old Zac Vawter successfully uses a nervous system-
controlled bionic leg to climb the Chicago Willis Tower.