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What is a
bioreactor?
A bioreactor may refer to any manufactured or
engineered device or system that supports a
biologically active environment.
In one case, a bioreactor is a vessel in which a
chemical process is carried out which involves
organisms or biochemically active substances
derived from such organisms.
This process can either be aerobic or anaerobic.
These bioreactors are commonly cylindrical,
ranging in size from liters to cubic meters, and
are often made of stainless steel.
A bioreactor may also refer to a device or system
meant to grow cells or tissues in the context of
cell culture.
Background
The first microbial bioreactors, in
particular
Escherichia
coli
and
Saccharomyces cerevisiae, were found to
be satisfactory for the production of
simple polypeptides such as insulin and
human growth hormone.
However, microbial bioreactors were
found to be unsuitable for proteins with
complex post-translational modifications
or intricate folding requirements, such as
the coagulation factors, or monoclonal
antibodies
Animal bioreactors
Animal systems for production
Blood
Urine
Seminal plasma
Egg white
Silk worm cocoon
Milk
Transgenic Animal- A
suitable Bioreactor
The animal bioreactor refers to an animal
with
bacteria in its digestive tract. The
bacteria may be a modified bacteria.
The animal may be a cow, pig, goat, sheep,
rabbit, horse, mouse, rat or guinea pig.
The bacteria may be present in the lumen
or the rumen, in the case of a ruminant
animal.
The bacteria may comprise a plasmid with a
heterologous nucleic acid, which may be
operatively linked to a regulatory element.
Preparation of an Animal
Bioreactor
Any animal with a digestive tract that is
capable
of
supporting
an
enteric
microorganism may be used as a bioreactor.
The animals may include cows, pigs, goats,
sheep, rabbits, horses, mice, rats and guinea
pigs.
The animals must be healthy and germ free.
The germ free host animals may be useful for
producing and maintaining the desired levels
of microorganisms in the digestive tract of
the animal fermentation chamber.
Administration of
Microorganisms
Animal bioreactors may be produced by
administering a microorganism to the host
animal by any method which allows for
introduction of the microorganism to the
digestive tract of the host animal including,
oral, nasal, and rectal administration.
The microorganism may be formulated in
any manner which allows for introduction
and propagation of the microorganism in
the digestive tract of the host animal
including
liquid
cultures,
lyophilized
cultures, encapsulated cultures, and agar.
Chemical
methods
for
disrupting
microbial cell walls include treatment
with
alkali,
organic
solvents
or
detergents.
If the product of interest is stable at
about pH 10.5-12.5, lysis may be carried
out on a large scale at low cost.
Lysis may also be performed using
enzymatic treatments which may be
highly specific and which may be
performed under mild conditions.
After cell disruption, cell debris is
removed by methods including lowspeed, high-capacity centrifugation or
membrane microfiltration.
Transgenic Sheep as
Bioreactor
The research in producing transgenic sheep
is now focused on producing sheep with
better
growth,
increased
meat
and
developing the mammary gland of this
mammal as a bioreactor.
The pharmaceutically important proteins are
made to secrete into the milk generated by
the sheep.
Though the amount of milk compared to
cattle is
less, yet lactation in sheep can produce
significant amount of milk on annum basis.
Applications of Transgenic
Sheep
1) Human growth hormone has been
successfully introduced into sheep in order
to increase their
development, growth and meat
production. Such
transgenic sheep has shown
considerable improvement in their body
weight, feed efficiency, meat/fat ratio and
fat composition. The gene for ovine growth
hormone is usually placed under control of
metallothionein promoter1
Transgenic Chickens as
Bioreactors
The hen since from long time has been a
potential
candidate
to
produce
human
biopharmaceuticals at low-cost with high-yield.
The reason for this is simple as:
The yolk and white of the egg are sterile.
The technology for fractionating egg yolk and
egg white proteins is available.
Highly
automated
systems
for
efficiently
producing and collecting thousands of eggs per
day are well established.
The egg white contains ~4 g of protein, more
than half of which comes from the expression of
a single gene i.e. ovalbumin gene (OV).
Hence, the OV promoter, combined with its other
Above
all
they
show
posttranslational modifications that can
be compared with that taking place
in humans.
The ability to express transgenes in milkproducing animals has resulted in the creation
of bioreactors.
These are animals that produce large amounts
of a given recombinant protein in their milk.
These recombinant proteins are produced in
fully biologically active form through proper
posttranslational
modification
(PTM),
for
purification and therapeutic use.
This approach has been used to produce
recombinant tissue plasminogen activator ,
granulocyte colony-stimulating factor, Ig , and
lactoferrin in the milk of goats and cows.
Cow
Gestation period (months)
1
9
Sexual maturity (months)
5
15
Time from introduction of transgene
to the beginning of lactation (months)
Pig
Sheep
Goat
Female Founder
Lactation induced in puberty
Natural Lactation
7
33
Male Founder
Lactation induced in puberty
(daughters)
45
Natural Lactation (daughters)
15
57
Average progeny
8
1
Annual Yield of milk production
(L/year)
4-5
8000
Production of the recombinant protein
/female/year (kg)
0.02
40
16
18
28
10
18
22
31
1-2
300
500
1.5
2.5
22
31
1-2
800
Animal bioreactors
Exemplary categories of polypeptides
Growth factors
Hormones
Antiviral proteins
Lipocortins
Lipotropins
Interleukins
Interferons
Stimulating factors
Kinases
Transmembrane
regulators
Immunoglobulins
Milk lipases
Cell surface proteins
Human pancreatic enzymes
Enkephalins
Silk proteins
Spider silk proteins
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